Characterization of sacha inchi (Plukenetia volubilis)&nbsp;and taro (Colocasia esculenta) flours with&nbsp;potential application in the preparation of both gluten-free and high protein foods.

Ruby Aracely Narváez Cadena; Angie Paola Salas Zambrano; Jesús Eduardo Bravo Gómez; Karen Sofia Muñoz Pabon; Diego Fernando Roa-Acosta

doi:10.12688/f1000research.130394.2

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Research Article

Revised

Characterization of sacha inchi (Plukenetia volubilis) and taro (Colocasia esculenta) flours with potential application in the preparation of both gluten-free and high protein foods.

[version 2; peer review: 2 approved with reservations, 2 not approved]

Ruby Aracely Narváez Cadena¹, Angie Paola Salas Zambrano¹, Jesús Eduardo Bravo Gómez¹, Karen Sofia Muñoz Pabon ¹, Diego Fernando Roa-Acosta¹

Ruby Aracely Narváez Cadena¹, Angie Paola Salas Zambrano¹, [...] Jesús Eduardo Bravo Gómez¹, Karen Sofia Muñoz Pabon ¹, Diego Fernando Roa-Acosta¹

PUBLISHED 09 Oct 2023

Author details Author details

¹ Departamento de Agroindustria, Universidad del Cauca, Popayan, Colombia

Ruby Aracely Narváez Cadena
Roles: Formal Analysis, Investigation, Methodology

Angie Paola Salas Zambrano
Roles: Formal Analysis, Investigation, Methodology

Jesús Eduardo Bravo Gómez
Roles: Investigation, Resources, Supervision, Validation

Karen Sofia Muñoz Pabon
Roles: Data Curation, Formal Analysis, Writing – Original Draft Preparation

Diego Fernando Roa-Acosta
Roles: Conceptualization, Project Administration, Supervision, Validation

OPEN PEER REVIEW

REVIEWER STATUS

Abstract

Background: Interest in alternative sources of tubers and legumes has increased in recent years because of the constant search for raw materials that provide bioactive compounds with antioxidant potential benefits for consumers. The functionality of new raw materials is sought through physical and/or chemical modifications to develop and innovate new foods. The objective of this study was to characterize taro (TF) (Colocasia esculenta) and sacha inchi (Plukenetia volubilis) flours, obtained by the wet (SIF-WM) and defatted method (SIF-DM), as an alternative for the formulation of new functional foods.
Methods: The free polyphenols of the different mixtures were analyzed, and the antioxidant properties of the extracts obtained were measured using ABTS^.* (2,2′-Azino-bis(3-ethylbenzthiazoline-6-sulfonic acid). The ABTS radical method, which reacts with the phenolic compounds of the food matrix, using Trolox as a standard.
The blends were subjected to pasting analysis, flow profile tests, determination of viscoelastic properties (temperature sweep). Characterization of common microorganisms in these foods was performed.
Results: The highest protein value was obtained in the sacha inchi flour obtained by the defatted method (72.62). The majority of components in taro were carbohydrates (85.4%). About antioxidant and determination of free polyphenols, taro flour obtained values of 2.71 µmol ET/g and 7.47 mg EAG/g, higher than Sacha inchi flours. In the rheological analysis (pasting properties, flow profile, and viscoelasticity), we observed that adding taro flour in different mixtures increases the viscosity peak and a lower breakdown, while there was an increase in setback. Except for defatted sacha flour, the others presented a flow index >1 before heating, showing a dilatant fluid behavior. The presence of Taro flour improves gel formation and stability. Staphylococcus aureus and Salmonella were present in the taro flour.
Conclusions: The flours analyzed represent a raw material with great potential for the development of gluten-free foods with functional properties.

Keywords

Rheology properties, Proximal composition, Microbiological quality, Gluten-free foods

Corresponding author: Karen Sofia Muñoz Pabon

Competing interests: No competing interests were disclosed.

Grant information: This work has been fully financed by the Colombian General Reimbursement System, the Uni-versity of Cauca, and SEGALCO S.A.S., within the framework of the SGR BPIN 2020000100052 project.
The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Copyright: © 2023 Narváez Cadena RA et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

How to cite: Narváez Cadena RA, Salas Zambrano AP, Bravo Gómez JE et al. Characterization of sacha inchi (Plukenetia volubilis) and taro (Colocasia esculenta) flours with potential application in the preparation of both gluten-free and high protein foods. [version 2; peer review: 2 approved with reservations, 2 not approved]. F1000Research 2023, 12:378 (https://doi.org/10.12688/f1000research.130394.2) First published: 06 Apr 2023, 12:378 (https://doi.org/10.12688/f1000research.130394.1) Latest published: 07 Mar 2025, 12:378 (https://doi.org/10.12688/f1000research.130394.4)

Revised Amendments from Version 1

In the new version, the figure of the sacha inchi defatting process is added.
Units are verified and adjusted, paragraph 1 of the introduction is rewritten. All the comments requested by the reviewer are adjusted.

See the authors' detailed response to the review by Lorena Sciarini
See the authors' detailed response to the review by Elisa Julianti
See the authors' detailed response to the review by Umi Purwandari
See the authors' detailed response to the review by Diego Salazar
See the authors' detailed response to the review by Oluwaseun P Bamidele

Introduction

According to Ref. 1, by 2050, if governance allows for debate and equity in global food and if food demand is very high, driven by rapid population growth, a high intensity of land use will be required to meet the demand for food, mainly meat and dairy products that are important sources of protein, vitamins and minerals in the human diet.² However, intensive livestock production is unsustainable from an environmental point of view, due to the demand for resources such as land to produce the crops needed for animal feed and fresh water; in addition, animal production is associated with the generation of greenhouse gases, a situation that contributes to the acceleration of global warming.³ Therefore, it is possible to achieve an adequate protein intake through the right combination of proteins of vegetable origin.

In this scenario, we must design food systems to contribute to global environmental sustainability and meet nutritional needs, proposing diets based on plants that allow including adjustments according to each region coupled with cultural customs.⁴ Underutilized or orphan crops are used to design original foods, not aimed at international trade, however, because of their high adaptation to the local environment, these crops play an important role in regional nutritional security.⁵ The mixture of plants such as cereals, pseudocereals, legumes, tubers, among others, are used to create different products with adequate balance of protein, carbohydrates, fiber and micronutrients such as vitamins and minerals.

For example, sacha inchi (Plukenetia volubilis) known as wild peanut, Inca peanut, sacha peanut or mountain peanut, is an oleaginous plant that belongs to the Euphorbiaceae family.⁶ Today, sacha inchi is still cultivated in the lowlands of the Peruvian Amazon and has been planted for centuries by the indigenous population.⁷ In Colombia, according to Ref. 8, production exceeds 2,400 tons of sacha inchi seed. The department of Putumayo is the largest producer of sacha inchi with 282 hectares, followed by Valle del Cauca, Caquetá and Antioquia.

Sacha inchi is a promising and industrializable food native to the Amazon that has essential unsaturated fatty acids such as omega-3, as alpha-linoleic acid with 47.7% to 51.9% and omega-9 as oleic acid with 7.9 to 8.9% by weight of oil, 27.4% protein, 4% ash and about 50% oil.⁹ These nutritional properties make sacha a favorable food for health and a suitable crop for developing high-protein and gluten-free foods.

Taro is a tuber abundant in starch of which 17-28% is amylose and the remaining is amylopectin.¹⁰ Taro has a high content of resistant starch that allows slow digestion with valuable effects on cholesterol and blood glucose regulation.¹¹ Taro has a high carbohydrate (59.36%) and protein (24.99%) content, with respect to the mineral content of taro, the highest amount present is magnesium 242.373 mg/kg, followed by calcium 94.455 mg/kg, iron 8.351 mg/kg and zinc 6.210 mg/kg, and, in relation to vitamins, is reported content of vitamin C 0.188 mg/100 mg, vitamin B1 0.047 mg/100 mg, and vitamin B3 0.078 mg/100 mg.¹² Compared to other tubers such as sweet potato, potato, cassava and yam; taro has a higher protein and fat content.¹² Likewise, taro has flavonoids and phenolic acids that have antioxidant properties; flavonoids, the largest group of phenolic compounds identified in the whole plant, are associated with reducing many degenerative diseases.¹¹

Foodstuffs are complex systems of great nutritional richness and therefore sensitive to attack by microorganisms (bacteria, fungi and yeasts). The main mode of contamination of raw materials is animal defecation, manure fertilization and recontamination persisting in facilities and transport; in addition, insects and rodents are a source of contamination.¹³ The low water activity (a_w<0.60) of meal does not favor microbial growth; however, contaminating spores along with inactive microorganisms will remain viable for prolonged periods and constitute a potential health hazard. As a persistence mechanism, Salmonella and other pathogens form biofilms that protect against disinfection and increase their tolerance to drying processes.¹³

In food matrices there is always a microbial load that must not exceed certain limits, according to Colombian regulations, which must be controlled to avoid the deterioration of the product and the consequent loss of its quality and suitability for consumption.

The design and production of new foods that respond to food and nutritional security in the midst of climate change in low-income countries is a challenge, so this study aimed to characterize the chemical, microbiological, rheological and bioactive properties of individual flours and mixtures of taro and sacha inchi, as potential foods applied in the formulation of different foods.

Methods

The taro tubers were bought in the Municipality of Orito Putumayo, located at 0° 38′ North Latitude and 76° 37′ West Latitude of Greenwich. Average temperature of 25 °C and relative humidity of 88%. The fresh Sacha inchi almonds were supplied by the company Fruty Amazónicos S.A.S. located in the village of La Concordia in the municipality of Valle del Guamuez, located at 00° 25″ north latitude and 76° 54″ west longitude. Temperatures range between 27 °C and 40 °C.

Obtaining taro flour

The taro flour (TF) was obtained following the methodology of Ref. 10. The raw material was received, the peel was removed; it was washed to eliminate impurities and then the edible part was cut into slices to facilitate dehydration; then it was weighed on a FENIX Electronic Weight Only Table Scale to verify yield, subsequently it was dried in a rotary oven (ORVES, Colombia) at a temperature of 60 °C for five hours to eliminate excess moisture. The dried slices were then ground in an electric mill (Quaker City Mill, model 4-e, Philadelphia) and then passed through a 30-mesh sieve system to achieve homogeneity in flour particle size.

Obtaining Sacha inchi flour

Sacha inchi seeds were received in white kernel, i.e., without shell or husk. The flour was obtained using the two methods described below:

Wet method

Sacha inchi flour from the wet method (SIF-WM), was obtained as follows: wet milling was performed in a blender (Oster, BLST 4655, Colombia) of 1.25 L, to which the sacha inchi and water were added in a 1:3 ratio, this, in order to disintegrate the grain. Obtaining a milky and homogeneous suspension. In order to separate the insoluble extract (cake) from the water-soluble extract (milk), a cloth filter was used and then the cake was dried in an oven at a temperature of 65 °C for 3 hours. Once dehydrated, it was immediately packed in a hermetically sealed polyethylene bag.¹⁴

Defatted method

The defatted sacha inchi (SIF-DM) flour was obtained as follows: the sacha inchi kernel was placed in the hopper of the automatic tactile oil extraction press (CGLDENWALL, store model K28, Shanghai China), as shown in Figure 1, a temperature of 124 °C was used. The press used the screw, which pushed the material into the main pressing cylinder, the pitch of the screw and the depth of the spiral were reduced, decreasing the volume available in the chamber, and the material being subjected to high pressure accompanied by friction against the closed bottom wall of the cylinder produced the extraction of the oil. The oil then flows through the existing orifices in the cylinder. The remaining material exits through the pressing system and the cake outlet nozzle.

Figure 1. Sacha inchi defatting process.

Proximate analysis

The proximate composition (protein, lipids, dietary fiber, ash and moisture) of taro and sacha inchi flours was determined according to the methods proposed by the AOAC (Association of Official Analytical Chemists, 1990)¹⁵ and the carbohydrate content was estimated by difference. The details of the method are explained in more detail in the protocols uploaded in the repository.

Water activity measurement

To determine the a_w of the raw materials, the powdered sample was placed in a Decagon Pawkit portable water activity meter until the cuvette was covered. Before reporting the measurement, the equipment was calibrated with standard salts at 0.25 (13.41 mol/Kg LiCl 0.25 a_w) and 0.76 (6.00 mol/Kg NaCl 0.76 a_w).

Protein

On nitrogen-free paper, 0.2 to 0.8 g of sample plus 1 g of kjeldahl catalyst were weighed and placed in digestion tubes. 10 mL of sulfuric acid were added. Then, gentle heating was initiated until no foaming or splashing was observed, using a temperature ramp as follows: 125 °C for 30 min, 270 °C for 30 min and 400 °C for 140 min.

The samples were digested until they were completely clear and translucent, free of organic matter, in the laboratory Kjeldahl digester (Raypa MBC-6/N, Spain).

Then they were cooled to room temperature in the Raypa distillation unit, after which each sample was titrated with 0.1 N HCl.

Lipids

For lipid testing, 1 g of sample was weighed in the extraction cartridges, then 80 mL of petroleum ether was added, immediately transferred in the rack to the laboratory Soxhlet and Randall Extractor (SX-6MP, RAYPA, Spain). After the extraction time and solvent recovery, the samples were placed in an oven at 60 °C for 1 hour to eliminate the remaining ether.

Fiber

For fiber testing, 1-2 g of the degreased sample were transferred to the laboratory fiber extractor (F-6P Fibertest, Spain) and fixed to the angle of the front part of the unit. Then 150 mL of 0.255 N H₂SO₄ was heated in an Erlenmeyer flask, when it was boiling, it was placed on top of the coolant, the heating knob was adjusted to boiling point 3 or 4 and left boiling for 30 minutes. At the end of this time, it was filtered and washed with distilled water and the operation was repeated 3 times using 30 mL of water each time. Then, 150 mL of sodium hydroxide solution preheated to 90 °C was placed in the upper part of the cooler; it was brought to boiling and kept for another 30 minutes. Then it was filtered and washed three times with boiling water. The sample was then placed in an oven at a temperature of 100-110 °C until a constant weight was obtained.

Determination of free polyphenols, FPP fraction

The determination of polyphenols was performed according to Ref. 16, with slight modifications. Two grams of the sample were weighed in a 50 mL falcon tube. The first fraction was added to the sample 8 mL of solution with 80% ethanol and 20% water and 1% formic acid (80 mL ethanol + 20 mL water + 1 mL formic acid). The samples were shaken for 25 minutes in a shaker at 200 RPM at room temperature. The samples were then centrifuged at 3500 rpm for 5 minutes at room temperature. Subsequently, the supernatant was taken in a new falcon tube (50 mL) and then 40 microliters of EDTA 2% was added to the supernatant.

For the second extraction, 8 mL of 70% acetone with 1% formic acid (70 mL acetone + 30 mL water + 1 mL formic acid) was added to the pellet of the first extraction. The pellet was shaken in solution for 25 minutes in the shaker at 200 RPM at room temperature. Subsequently, the samples were centrifuged at 3500 rpm for 5 minutes at room temperature and the second supernatant was poured into the first supernatant and mixed. Finally, the supernatant mixture was made up to 20 mL using distilled water and the absorbance was measured in the spectrophotometer (Thermo scientific, Genesys 10S UV VIS).

Determination of antioxidant activity by the ABTS method

In a test tube 4 mL of ABTS.* (2,2′-Azino-bis(3-ethylbenzthiazoline-6-sulfonic acid), solution was added and to start the reaction 135 μL standard solution and vortexed for 5 seconds. A reagent blank was carried with 4 mL of 4.5 acetate buffer and 135 μL of ethanol, point 0 was prepared by adding 4 mL of ABTS solution and 135 μL of ethanol. Finally, the tube was capped and waited exactly 30 minutes to measure the absorbance in the spectrophotometer (Thermo scientific, Genesys 10S UV VIS) at a wavelength of 729 nm.

Rheological analysis

Pasting properties

The pasting properties of each dispersion were determined using a rheometer (TA INSTRUMENTS, AR 1500, New Castel, USA), equipped with a starch pasting cell. Then, a suspension of flour in water with a concentration of 12% (w/w) in 25 g was prepared and exposed to heating and cooling. The following samples were analyzed: 100%TF; 100%SIF-DM; 100%SIF-WM; 25%SIF-DM:75%TF; 50%SIF-DM:50%TF; 75%SIF-DM:25%TF; 25%SIF-WM:75%TF, 50%SIF-WM:50%TF; 75%SIF-WM:25%TF. The shear rate was kept constant at 16.75 s^-1, throughout the heating and cooling range (25 °C–90 °C–25 °C) while the heating rate was 10 °C/min. The following parameters were obtained from the rheological analysis maximum viscosity [Pa.s]; minimum viscosity [Pa.s]; breakthrough viscosity [°C] and setback viscosity [Pa.s]. Finally, using the Savistky-Golay function, the data were smoothed in the GraphPad Prism 8.0.1 program (RRID:SCR_002798).¹⁷^,¹⁸

Flow profile

Flow properties were performed according to the methodology of Ref. 17 with modifications. An AR1500 rheometer (TA Instruments, New Castel, USA) was used in this study. The average viscosity was determined at 25 °C for 12 min with a shear rate increasing in 4 steps as follows:

Step 1: 1×10^-3 s^-1 to 1×10^-2 s^-1

Step 2: 0.01 s^-1 to 0.1 s^-1

Step 3: 0,1 s^-1 to 1 s^-1

Step 4: 1 s^-1 to 100 s^-1

Subsequently, the sample was heated to 90 °C. Once this process was completed, the average viscosity was determined again at 25 °C, subjecting the sample to the same shear rate.

Flow curves (shear stress versus shear rate) were obtained and fitted to the power law model shown in Equation 1.

(1)

T = K * y^{n}

Where:

T is the shear stress (Pa).

y is the shear velocity (s^-1)

K is the coefficient of consistency (Pa. s^-1)

n is the flow behavior index

The flow behavior index indicates Newtonian flow behavior when n = 1, shear thinning behavior when n < 1 and shear thickening when n > 1). Consistency and creep were determined before and after heating in order to determine the effect of thermal processing on these parameters.

Determination of viscoelastic properties (temperature sweep)

For the determination of viscoelastic properties (temperature sweep) the method of Ref. 19 was used, the viscoelastic properties were determined in a rheometer (TA INSTRUMENTS, AR 1500, New Castel, USA) using a system of parallel flat plates, of 40 mm diameter and 1500 μm distance between plates. The edges of the plates were sealed with vaseline, with the purpose of controlling evaporation and avoiding variations in concentrations of the aqueous suspensions used.

The samples were subjected to a cycle of dynamic heating (25-85 °C), stabilization (85 °C for 2 minutes) and cooling (85-25 °C) at 10°C/min, with a frequency of 0.5 Hz and 0.5% deformation. The profiles of the viscoelastic moduli as a function of temperature were recorded using the equipment’s software.

Microbiological characterization

The microbiological characterization of the flours was carried out, considering the methodology reported by Ref. 20, which was based on parameters established by the Colombian technical standard NTC that applies for each microorganism.

To perform the microbiological characterization, 10 g of sample were taken in triplicate, then diluted in 90 mL of peptonized water and mixed at 150 rpm for 10 min in a shaker (MaxQ 4450 orbital Thermo Ficher Scientific USA), following this, for each microorganism the seeding of the appropriate dilution and selective medium is described. The details of the method are explained in more detail in the protocols uploaded in the repository https://doi.org/10.5281/zenodo.7582202.

Coliforms

1 mL of each replicate was taken and seeded in a previously sterile Petri dish by seeding in depth on chromogenic colinstant agar for 24-48 h of incubation at 35 °C.

Mesophiles

1 mL of sample was taken from each replicate, and each dilution was seeded by immersion in previously sterile Petri dishes in Plate Count Agar (PCA) agar for 24-72 h at 30 °C.

Fungi and yeasts

100 μL of each replicate was taken and seeded per surface in previously sterile Petri dish by surface plate seeding on Potato Dextrose Agar (PDA) agar for 24-72 h incubation at 30°.

Bacillus cereus

100 μL of each replicate was taken and surface seeded in previously sterile Petri dish by surface plate seeding on Mannitol egg Yolk Polymyxin agar (MYP) agar with egg yolk specific for Bacillus cereus for 24 h incubation at 37 °C.

Staphylococcus aureus

100 μL were taken from each replicate and seeded per surface in a previously sterile Petri dish, by surface plate counting on Baird Parker agar at 24 h incubation at 35 °C, and then incubated for 24 h at 35 °C.

Salmonella

25 g of sample were taken in duplicate, then diluted in 225 mL of buffered pepton water and mixed at 150 rpm for 10 min in shaker (MaxQ 4450 orbital Thermo Ficher Scientific USA), once this time was over, they were left in incubation at 37 °C for 18 h, after which 100 μL were inoculated in 10 mL of Rappaport Vassiliadis malachite green broth (RVS medium) at 41. Furthermore, 1000 μL were inoculated in 10 mL of Tetrationate Mueller Kauffmann broth (MKTTn medium) at 37 °C for 24 h. After this time, the surface of a Petri dish containing XLD agar selective medium was seeded by means of a loop in such a way that asylated colonies were obtained, Salmonella Shiguella agar was incubated at 34 °C for 24 h, at the end of which typical colonies of Salmonella spp. were observed.

Statistical analysis

A completely randomized design (nine treatments) was used to evaluate the rheological properties of native and formulated flours. The design was based on the type of flour and its inclusion levels in the mix, which were: 100% TF; 100%SIF-DM; 100%SIF-WM; 25%SIF-DM:75%TF; 50%SIF-DM:50%TF; 75%SIF-DM:25%TF; 25%SIF-WM:75%TF; 50%SIF-WM:50%TF; 75%SIF-WM:25%TF. The response variables were viscosity as a function of temperature-shear rate and viscoelastic modulus (G′ and G″).

The results were presented as the mean ± standard deviation of triplicate experiments. One-way analysis of variance (ANOVA) was used to compare means. Differences between means were considered significant at P < 0.05 using Tukey’s new multiple range test. Data were subjected to analysis with Minitab version 20 (RRID:SCR_014483), https://www.minitab.com/es-mx/support/downloads/. Graphs were generated in Graphpad Prism 5.0.

Results and discussion

Proximate analysis

Table 1 shows the results of the proximate analysis for the samples. Plukenetia volubilis seed on dry basis has a high lipid content (52.84%) like oilseeds such as soybean (16.61-24.71%), almond (Prunus dulcis) and peanut (Arachis hypogaea) whose values vary around 50%.²¹^,²²

Table 1. Proximate composition of flours.

Component	Sacha inchi almond	SIF-WM	SIF-DM	TF
% Protein	33.73^b ± 0.01	31.54^c ± 0.04	72.62^a ± 0.10	6.05^d ± 0.13
% Ashes	3.05^c ± 0.001	2.56^d ± 0.06	6.79^a ± 0.04	5.18^b ± 0.01
% Lipids	52.84^a ± 0.62	8.87^c ± 0.27	9.84^b ± 0.24	0.70^d ± 0.05
% Crude fiber	3.71^c ± 0.18	19.42^a ± 0.32	7.71^b ± 0.01	2.65^d ± 0.16
% Carbohydrates	6.67	37.61	3.04	85.42

The acronym (TF) refers to taro flour; (SIF-WM) sacha inchi flour by wet method and (SIF-DM) sacha inchi flour by defatted method.

Values are presented as mean ± SD. For each parameter, different letters indicate significant differences at p < 0.05.

The results of lipids are within the range of values reported by Refs. 23–26. The protein content of sacha inchi kernel (33.73%) is above the range, but close to the value reported by Ref. 24. As for ash, the value was 3.05%, crude fiber (3.71%), and carbohydrates (6.67%). Regarding moisture, sacha inchi seed presented a 6.62%, close to the value reported by Ref. 26 with 6.28%.

Two methods were used to obtain sacha inchi flour: the wet method and mechanical defatting.

Wet method: The flour obtained by the wet method had an oil content of 8.87% and a protein content of 31.54%, with a protein/fat ratio of 3.56. The high protein values obtained from sacha inchi cake using this mechanical pressing method are the basis for the formulation of protein-rich foods using vegetable sources.

Mechanical defatting method: the sacha inchi flour obtained by the mechanical defatting method had a protein content of 72.62%, being above the range of values reported by Refs. 23, 27–30 which vary between 41.49 and 65.6%.

Crude fiber has a value of 7.71%, the carbohydrate content is 3.04%, below that reported by Ref. 27 with a value of 11.25%. This difference can result from suspended solids carried in the oil stream.

The comparison of the two methods for obtaining sacha inchi flour (SIF-DM and SIF-WM) made it possible to see the method that presented the highest protein content was SIF-DM with 72.62% compared to a value of 31.55% for SIF-WM. The lipids results were close, 9.84% for the SIF-DM and 8.87% for SIF-WM. As for fiber, SIF-WM presented higher values (19.42%) compared to SIF-DM (7.71%).

Taro flour: as shown in Table 1, carbohydrates are the major component of taro with a value of 85.42%. The values of protein (6.05%), ash (5.18%), and lipids (0.70%) from proximate analysis agree with research.¹⁰^,³¹^–³⁵ Crude fiber is the only data below the range consulted with a value of 2.65%, the closest value being that reported by Ref. 35 with a value of 4.38%.

Effect of processing on FPP and the antioxidant activities

Table 2 shows that the total polyphenols obtained in the flours are statistically different, taro flour presents the highest polyphenol content of 7.50 ± 1.73 mg GAE/g, being like those reported by Ref. 36, in comparison with flours from other sources such as plantain (5.93 ± 0.43 mg GAE/g)³⁷ present a higher value. The defatted sacha inchi flour presented a polyphenol content of 3.37 ± 0.16 mg GAE/g flour, being higher than the flour obtained by the wet method, because the mechanical extraction involving pressure and temperature allows a release of phenolic compounds.³⁸

Table 2. Bioactive properties of papachina and sachainchi flour.

Raw material	FPP (mg GAE/g sample)	ABTS (μmol ET/g sample)	a_w
TF	7.47^a ± 0.27	2.71^a ± 0.02	0.61
SIF-WM	2.68^c ± 0.26	0.49^b ± 0.02	0.64
SIF-DM	3.37^b ± 0.16	0.71^c ± 0.04	0.54

The acronym (TF) refers to taro flour; (SIF-WM) sacha inchi flour by wet method and (SIF-DM) sacha inchi flour by defatted method; (FPP) refers free polyphenols.

Values are presented as mean ± SD. For each parameter, different letters indicate significant differences at p < 0.05.

Antioxidants present in foods play an important role in their preservation because they prevent oxidation processes; in addition, these compounds contribute to health promotion through the prevention of pathological processes mediated by oxidative stress. Therefore, the formulation of new foods that present bioactive compounds is attractive for the new trends in the design of new foods.

Regarding the antioxidant activity measured by the ABTS method, taro flour obtained a statistically higher antioxidant activity with 2.71 ± 0.02 μmol ET/g compared to SIF-DM (0.49 ± 0.02 μmol ET/ g) and SIF-WM (0.71 ± 0.04 μmol ET/g). This difference can be explained because taro is a prime source of antioxidant compounds and, from a chemical point of view, has various biologically active phytoconstituents, such as flavonoids, sterols, and glycosides.³⁹

In this study, a TF drying temperature of 60 °C was used, which according to Ref. 40, temperatures between 50 °C and 60 °C allow for maximum antioxidant activities in taro flours. SIF-DM presented the lowest antioxidant activity, possibly due to the fact that during extraction the oil stream carries away bioactive compounds, which in sacha inchi are present particularly in polyunsaturated essential fatty acids, phytosterols and tocopherols, which are divided into three chemical categories: α-linolenic acid, β-sitosterol and γ- and δ-tocopherols.⁴¹

Rheological analysis

Pasting curves

The viscosity profile was carried out to know the evolution of viscosity during the heating and cooling phase in the raw materials and in each of the blends. According to the statistical analysis, the different flour blends significantly affect viscosity p < 0.05. Figure 2A shows the significant differences between the blends in the viscosity peak, according to the post hoc Tukey analysis five different groups identified with letters from “a” to “e” are classified. In group “a”, the samples with the highest viscosity (25%SIF-DM:75%TF and 100%TF) with values of 3.08 Pa.s and 2.99 Pa.s respectively, and in group “e”, the samples with the lowest viscosity value (75%SIF-WM:25%TF and 100%SIF-WM) with values of 0.35 Pa.s and 0.64 Pa.s respectively.

Figure 2. Effect of the different blends on the curves of the pasting curves.

The data represent the significant differences in the values of: (A) viscosity peak [Pa.s]; (B) Trough [Pa.s]; (C) Breackdown [°C] and (D) setback [Pa.s]. The acronym (TF) refers to taro flour; (SIF-WM) sacha inchi flour by wet method and (SIF-DM) sacha inchi flour by defatted method.

According to the results, the samples with higher TF content present the highest viscosity peaks, due to the presence of starch contributed by TF, taking into account that in this study taro, presented 85.42% of carbohydrates that according to Ref. 42 are represented by starch. Starch granules are not soluble in cold water, because of the strong hydrogen bonds that hold the starch chains together. Starch heated in excess water and above the pasting temperature suffers an order-disorder phase transition called gelatinization. This transition is associated with water diffusion in the granule, water absorption by the amorphous region, hydration and radial swelling of the starch granules, leading to an increase in viscosity. As Figure 3 shows, the greater the presence of a starch source, the higher the viscosity peak. This positive correlation of starch content with peak viscosity is consistent with research on different commercial flours from cereals, pseudocereals, roots, and legumes.⁴³ Comparing SIF-DM flour with SIF-WM, the defatting method produces a flour with higher viscosity, due to the interaction of water with proteins and starch,¹⁸ taking into account that SIF-DM has 72.62% protein. It can be attributed that the lower viscosity of the sacha inchi flour obtained by the wet method is due to the large particle size of the flour, due to its high oil content, which is shown in Table 2; it was not possible to perform a size reduction and we worked with the granules resulting from the liquefaction process and subsequent drying. Therefore, the endogenous lipids present in taro flour form a kind of protective layer of the carbohydrate and protein agglomerate, preventing water absorption and subsequent swelling of the starch.⁴⁴

Figure 3. Pasting properties of flour blends.

The graphs show the flow profile behavior of: (A) SIF-DM; (B) SIF-WM; (C) TF; (D) 25%SIF-DM:75%TF; (E) 25%SIF-WM:75%TF; (F) 50%SIF-DM:50%TF; (G) 50%SIF-WM:50%TF; (H) 75%SIF-DM:25%TF; (I) 75%SIF-WM:25%TF. The acronym (TF) refers to taro flour; (SIF-WM) sacha inchi flour by wet method and (SIF-DM) sacha inchi flour by defatted method.

Figure 2B shows the significant differences in the samples behavior before the onset of retrogradation (trough or drop viscosity), which occurs at the end of the constant temperature section, before cooling begins. There are six different groups, identified with letters from “a” to “f”, where in group “a”, are the samples with the highest trough value of 25% 25%SIF-DM:75%TF, with values of 1.84 Pa.s and 1.83 Pa.s respectively, and in group “f”, are the samples with the lowest trough value of 75%SIF-WM:25%TF and 100%SIF-WM with values of 0.1 and 0.02 respectively.

According to the above, as shown in Figure 3, the samples that presented a higher viscosity drop are the samples with higher TF content, this is because the crystalline structure (amylopectin) that makes up the granules is altered, when amylose and amylopectin are hydrated individually in excess water, both macromolecules tend to reorganize in a similar structure to the one they had initially.⁴⁵

As shown in Figure 3, mixtures with higher inclusion of starch present in tf and less protein contributed by sacha inchi result in more free water for starch molecules to swell and solubilize presenting higher viscosity; on the contrary, at high proportions of protein its interaction with starch granules could stabilize the amylose leaching channel of starch granules, which resulted in lower swelling power and solubility.⁴⁶

Figure 2C shows the significant differences in the samples behavior in the breakdown stage. According to Ref. 47, the breakdown or stability is the difference between the viscosity peak and the trough in the constant temperature section and shows disintegration at a maintenance temperature (95 °C) during continuous shear; the lower the Breackdown value, the higher the anti-shear capacity. In this study, according to the post hoc Tukey analysis six different groups identified with letters from “a” to “f”. In group “a”, the samples with the highest breakdown value (25%SIF-DM:75%TF and 100%TF) with values of 1.24 Pa.s and 1.16 Pa.s respectively, and in group “f”, the samples with the lowest value (100%SIF-WM) with values of 0.040 Pa.s. It was evidenced that the samples with low starch content contributed by taro present higher stability, while the samples with higher starch content have lower capacity to support viscosity during prolonged heating, because of the starch behavior, in which the low molecular weight amylose separates from the starch granule and they collapse at constant temperature until the amorphous part is solubilized, decreasing viscosity.⁴⁸

Figure 2D shows the significant differences between the mixtures in the setback parameter, according to the post hoc Tukey analysis five different groups identified with letters from “a” to “e”. In group “a”, we find the samples with the highest setback (100%TF and 25%SIF-DM:75%TF and 100%TF) with values of 1.69 Pa.s and 1.60 Pa.s respectively, and in group “e”, the samples with the lowest values (75%SIF-WM:25%TF and 100%SIF-WM) with values of 0.13 Pa.s and 0.03 Pa.s respectively. The variable reorganization or setback is the indicator of the retrogradation and rearrangement of starch molecules during the cooling process that defines the reabsorption of soluble starch polymers and insoluble granular fragments during the cooling phase.⁴⁹ According to the results, blends containing lower taro content are less prone to retrogradation. As mentioned above, starch (amylose-amylopectin) contributed by TF with variations in temperature generates changes, e.g., amylose forms double helix associations of 40 to 70 glucose units, since amylopectin crystallization occurs by reassociation of the outermost short branches, i.e., both amylose and amylopectin are capable of retrogradation, although the amylopectin seems to be responsible for long-term quality changes in foods.⁵⁰ In taro starch, amylopectin contents are higher than amylose 66 and 34 % respectively,⁵¹ which is the reason for attributing higher retrogradation in flour blends with higher TF.

Flow profile analysis

Figure 4 shows the results of the flow analysis with the viscosity behavior before heating (blue line) and after heating (red line) under stress conditions (shear stress, Pa) and shear rate (s^-1). We observed the coefficients obtained by regression of the power model, where “n” is the flow index and “K” is the consistency index. According to the consistency index, we see that “K” increases in all samples except for SIF-WM, because of the viscosity gain, being “n” and “K” inversely proportional parameters.

Figure 4. Flow profile of flour blends.

Regarding the flowability index, we observed the behavior of the samples with the highest starch content TF were those that presented the greatest change in “n”, decreasing their flowability index represented by the gain in viscosity attributed to the behavior of the starch in taro.

Regarding the flow index, all the mixtures, except for SIF-DM, have a value of n > 1 before heating, showing that they have a dilettantish fluid behavior. In water and food starch suspensions, the dilatant or shear thickening behavior relates to the initial stiffness of starch granules to resist shear and to a high concentration of solids, resulting in particle swelling.⁵² SIF-DM presented an n of 0.7709, demonstrating a behavior of a pseudo-plastic fluid.

For the mixture’s behavior after heating, all the blends, except for SIF-WM, presented an n < 1, demonstrating a behavior of a pseudo-plastic fluid, i.e., that changes in temperature affect these mixtures behavior. This behavior occurs because, in the gelatinization process, the starch granules break, releasing amylose to the aqueous medium and, on cooling, these amylose chains align, forming networks that form gels or viscous suspensions.¹⁷ The n values for the SIF-WM were reduced to 1.064, exhibiting Newtonian behavior. The Newtonian flow shows that the viscosity is independent of the shear rate. According to Ref. 52, these flours could be suitable for formulating products such as beverages, where improving the nutritional content is required without affecting its viscosity during shearing.

Viscoelastic properties - Temperature sweeping

Figure 5 presents the moduli G′ (red color) and G″ (blue color) as a temperature function (°C). We observe the behavior of the elastic and viscous moduli for each mixture, evidencing an increase in the moduli as the temperature sweeps are performed. Except for graphs “A” and “H”, the mixtures behavior during heating is similar, since the two moduli increase their value, decreasing the difference between them and achieving a crosslinking, where the material ceases to have a viscous character, i.e., it stops being liquid and becomes elastic properties characteristic of a solid. This behavior is characteristic of materials undergoing liquid-solid phase transformations. When the molecules gain weight, in this case, when the starch granules interlock, the loss modulus G" decreases and the storage modulus G′ increases, reaching a point of equilibrium that is its material change of nature, its solidification.

Figure 5. Temperature sweeping, modules G′ and G″ of the flour blends.

The graphs show the loss modulus G" and storage modulus G' for: (A) SIF-DM; (B) SIF-WM; (C) TF; (D) 25%SIF-DM:75%TF; (E) 25%SIF-WM:75%TF; (F) 50%SIF-DM:50%TF; (G) 50%SIF-WM:50%TF; (H) 75%SIF-DM:25%TF; (I) 75%SIF-WM:25%TF. The acronym (TF) refers to taro flour; (SIF-WM) sacha inchi flour by wet method and (SIF-DM) sacha inchi flour by defatted method.

There is a progressive exudation of amylose in the swollen granules, where gel-like linkages are formed. Researchers have reported the effect of amylose to increase the firmness of gels during cooling as one of the initial causes of gel firmness. Thus, during the cooling stage, the moduli stay constant with the predominance of the elastic modulus.⁵³

The behavior of flour blends A and H corresponding to 100% SIF-DM and 75% SIF-DM: 25% TF, respectively, shows that the G′ and G″ moduli remain constant both during the temperature increase and during subsequent cooling, thus presenting greater stability to temperature variations. This behavior responds to the low starch content of these samples, since starch is key for the viscoelastic behavior of the flours, due to the interaction with water, generating absorption and swelling of the granule and, consequently, greater viscoelasticity. SIF-DM and 75%SIF-DM:25%TF blends did not present a gel point, however, the other blends presented a gel-like behavior, highlighting that the 50%SIF-DM:50%TF and 25%SIF-DM:50%TF blends presented the most elastic gel at a gel temperature of 60 °C in both cases, showing similar viscoelastic characteristics.

We suggest an optimum level of protein inclusion represented by sacha inchi between 25 and 50%, where the consistency of the gel increases and mixtures prepared above this range do not present a gel point, while lower values will have a more liquid behavior.

Microbiological analysis

Table 3 shows the results of the microbiological analysis performed on the three types of flour. For the microbiological analysis, we considered the NTC 6069⁵⁴ standard for quinoa flours and NTC 267⁵⁵ for wheat flour.

Table 3. Microbiological analysis.

Microorganism	Specification (log UFC/g)	Counts (log UFC/g)
Microorganism	Specification (log UFC/g)	TF	SIF-DM	SIF-WM
Total mesophilic aerobic count	5.47	4.21 ± 0.09	4.32 ± 0.09	3.94 ± 0.05
Staphylococos count	<2	2.38 ± 0.09	1.52 ± 0.10	2.44 ± 0.08
Molds and yeasts count	3.7	1.51 ± 0.10	1.60 ± 0.08	2.02 ± 0.06
Total coliform count	<1	Absence	Absence	Absence
Salmonella in 25 g	Absence	Presence	Absence	Absence

The acronym (TF) refers to taro flour; (SIF-WM) sacha inchi flour by wet method and (SIF-DM) sacha inchi flour by defatted method.

As can be seen in Table 3, the flours comply with the quality specifications required by Colombian regulations; however, as shown in Figure 6, Salmonella is present in the taro flour, possibly due to the persistence of this type of microorganism; drying this flour at 60 °C preserves some bioactive compounds but is not sufficient to eliminate it. According to Ref. 14, this type of pathogens acquire resistance to disinfectants and temperature because they form biofilms, which protect them and can remain in equipment, utensils and facilities, contaminating food. In addition, flours such as taro can be contaminated during harvesting or transportation.

Figure 6. Colonies seeding taro flour sample on XLD agar after 24 h.

A: MKTTn broth, C: RVA broth, seeding of taro flour sample on Salmonella-Shigella agar B: MKTTn broth, D: RVA broth.

Table 2 shows that in foods such as sacha inchi and taro flour, where a_w is less than 0.85, Salmonella and other pathogenic bacteria such as Staphylococcus can survive in a viable but non-culturable state for long periods of time due to their increased resistance to thermal processes.⁵⁶ Although the growth of microorganisms is not maintained at low water activity, foodborne bacteria and fungi can easily contaminate flour and survive for long periods of time with a low reproduction rate.²⁰

Conclusions

The method that allows for the highest concentration of protein in sacha inchi is mechanical defatting with a protein result of 73%, compared to that obtained by the wet method, which resulted in 31%. However, during the production of sacha flour with the mechanical method, bioactive compounds are lost, which is why lower values of total polyphenols and antioxidant activity are obtained, compared to taro flour.

The viscosity profile evaluation determined that the starch contained in taro provides good viscosity characteristics, being this an important ingredient for the development of foods such as creams, pastes, sauces, among others. However, mixtures with excess taro have a higher rate of retrogradation, presenting a disadvantage for food stability during storage.

Through the viscoelasticity analysis (temperature sweep), we found the optimum level of inclusion of defatted sacha inchi is between 25 and 50%, where a stable and consistent gel is presented.

Although taro’s drying temperature of 60 °C maintains polyphenols and antioxidant activity, it does not allow the inactivation of bacteria such as Salmonella, which has shown resistance to thermal treatment in recent years.

Developing foods using gluten-free orphan raw materials, especially taro and sacha inchi that are easily adapted to regions with food safety hazards, can be a good alternative to offer the market different gluten-free products, excellent source of protein, and high fiber content.

Future research can formulate foods from sacha inchi flour, which will allow obtaining high protein foods, responding to the trend of obtaining high protein foods from vegetable sources.

Salmonella contamination was present in the taro flour. In response, the facilities and equipment were washed and disinfected with a different product than usual, and again salmonella analyses were performed to guarantee its elimination.

Data availability

Underlying data

Zenodo: Sacha_article, https://doi.org/10.5281/zenodo.7625871.⁵⁷

This project contains the following underlying data:

- REOLOGY SAMPLES.zip
- • Flow profile analysis_folder containing data on the flow analysis of flours in different samples. (Folder contains data on flow analysis and statistical analysis of flours in different blends)
- • Pasting_containing curves and statistical data of pasting properties in different samples. (Folder containing curves and statistical data of pasting properties)
- • Viscoelasticity_containing curve data and statistical analysis of analyzed viscoelastic properties in different samples. (The folder contains the data of the curves and the statistical analysis of the viscoelastic properties analyzed)

Extended data

Zenodo: Sacha_article, https://doi.org/10.5281/zenodo.7625871.⁵⁷

This project contains the following extended data:

Protocols on proximate and microbiological analysis

- Bacillus cereus--protocol for the determination of Bacillus cereus present in food according to the Colombian regulations is presented.docx. (This file contains the microbiology protocol for the determination of Bacillus cereus according to Colombian regulations)
- Coliforms--protocol for the determination of coliforms present in food according to the Colombian regulations is presented.docx. (This file contains the microbiology protocol for the determination of Coliforms according to Colombian regulations)
- Fungi and yeasts--protocol for the determination of fungi and yest present in food according to the Colombian regulations is presented.docx. (This file contains the microbiology protocol for the determination of Fungi and yeasts according to Colombian regulations)
- Mesophiles-- protocol for the determination of mesophiles present in food according to the colombian regulations is presented.docx. (This file contains the microbiology protocol for the determination of Mesophiles according to Colombian regulations)
- Salmonella Spp-- protocol for the determination of Samonella present in food according to the Colombian regulations is presented.docx. (This file contains the microbiology protocol for the determination of Salmonella Spp according to Colombian regulations)
- Staphylococcus aureus--protocol for the determination of Salmonella present in food according to the Colombian regulations is presented.docx. (This file contains the microbiology protocol for the determination of Staphylococcus aureus according to Colombian regulations)
- Protocol Fiber--protocol for fiber determination is presented.docx. (This file contains the proximal analysis protocol for fiber determination and handling of the equipment used for this determination)
- Protocol Lipid--protocol for lipds determination is presented.docx. (This file contains the proximal analysis protocol for lipds determination and handling of the equipment used for this determination)
- Protocol Protein--protocol for protein determination is presented.docx. (This file contains the proximal analysis protocol for protein determination and handling of the equipment used for this determination.)

Data are available under the terms of the Creative Commons Attribution 4.0 International license (CC-BY 4.0).

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53. Paulet PG, Bello-pérez LA, Salas F: Características viscoelásticas y estimaciones de masas moleculares en almidón de oca (Oxalis tuberosum). Rev. la Soc. Química del Perú. 2009; 75(2): 266–276.
54. ICONTEC: Harina de trigo. NTC 267.2007.
55. ICONTEC: Productos de molinería. harina de quinua. requisitos. Instituto colombiano de normas técnicas y certificación. NTC 6069.2014.
56. Subedi S, Du L, Prasad A, et al.: Inactivation of Salmonella and quality changes in wheat flour after pulsed light-emitting diode (LED) treatments. Food Bioprod. Process. 2020; 121: 166–177. Publisher Full Text
57. Sofia K: Sacha_article. [Data]. 2023. Publisher Full Text

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Version 4

VERSION 4 PUBLISHED 06 Apr 2023

Author details Author details

¹ Departamento de Agroindustria, Universidad del Cauca, Popayan, Colombia

Ruby Aracely Narváez Cadena
Roles: Formal Analysis, Investigation, Methodology

Angie Paola Salas Zambrano
Roles: Formal Analysis, Investigation, Methodology

Jesús Eduardo Bravo Gómez
Roles: Investigation, Resources, Supervision, Validation

Karen Sofia Muñoz Pabon
Roles: Data Curation, Formal Analysis, Writing – Original Draft Preparation

Diego Fernando Roa-Acosta
Roles: Conceptualization, Project Administration, Supervision, Validation

Competing interests

No competing interests were disclosed.

Grant information

This work has been fully financed by the Colombian General Reimbursement System, the Uni-versity of Cauca, and SEGALCO S.A.S., within the framework of the SGR BPIN 2020000100052 project.
The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Narváez Cadena RA, Salas Zambrano AP, Bravo Gómez JE et al. Characterization of sacha inchi (Plukenetia volubilis) and taro (Colocasia esculenta) flours with potential application in the preparation of both gluten-free and high protein foods. [version 2; peer review: 2 approved with reservations, 2 not approved]. F1000Research 2023, 12:378 (https://doi.org/10.12688/f1000research.130394.2)

NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article.

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Key to Reviewer Statuses VIEW HIDE

ApprovedThe paper is scientifically sound in its current form and only minor, if any, improvements are suggested

Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit.

Not approvedFundamental flaws in the paper seriously undermine the findings and conclusions

Version 2

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Reviewer Report 17 Sep 2024

Umi Purwandari, University of Trunojoyo Madura, Madura, Indonesia

Approved with Reservations

https://doi.org/10.5256/f1000research.155702.r317396

The paper was written carefully enough and data were analyzed thoroughly using recent related publication. However, microbiological analysis did not seem relevant to the work, and therefore it would be better deleted.

The term 'flour size' was confusing. Was it starch granule size, and if it was so, did the authors measure it, or is that any supporting argument for judging the size.

Some parts were not correctly written, such as in page 10 where it was refer to Table2 for oil concentration. While Table 2 was about antioxidant activity. Also, statistical notation for Table 2, ABTS did not look right.

Is the work clearly and accurately presented and does it cite the current literature?

Partly
Is the study design appropriate and is the work technically sound?

Yes
Are sufficient details of methods and analysis provided to allow replication by others?

Partly
If applicable, is the statistical analysis and its interpretation appropriate?

Yes
Are all the source data underlying the results available to ensure full reproducibility?

Yes
Are the conclusions drawn adequately supported by the results?

Yes

Competing Interests: No competing interests were disclosed.

Reviewer Expertise: Food science, food microbiology

I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above.

CITE

Report a concern

Author Response 22 Nov 2024

Karen Muñoz, Departamento de Agroindustria, Universidad del Cauca, Popayan, Colombia

22 Nov 2024

Author Response

Dear Dr Purwandari, Thank you very much for your thoughtful and constructive feedback on our manuscript. We appreciate the time and effort you dedicated to reviewing our work, as well ... Continue reading Dear Dr Purwandari, Thank you very much for your thoughtful and constructive feedback on our manuscript. We appreciate the time and effort you dedicated to reviewing our work, as well as your insightful suggestions, which have greatly helped us improve our paper. Below, we address each of your comments in detail:

1.The paper was written carefully enough and data were analyzed thoroughly using
recent related publication. However, microbiological analysis did not seem relevant
to the work, and therefore it would be better deleted.
Regarding the microbiological analysis, it is important to note that this was conducted to
evaluate the initial state of the obtained flours concerning their microbiological
composition. Since these flours are intended for food consumption, it is essential to know
the level of microbial growth and to ensure that they fall within the permissible ranges
established by current regulations governing such products.

2. Some parts were not correctly written, such as in page 10 where it was refer to
Table2 for oil concentration. While Table 2 was about antioxidant activity. Also,
statistical notation for Table 2, ABTS did not look right.

With the purpose of addressing the question raised, the information was supplemented
using the correct term ;granulometry.; Additionally, the article mentions the following
textually “Comparing SIF-DM flour with SIF-WM, the defatting method produces a
flour with higher viscosity due to the interaction of water with proteins and starch,
considering that SIF-DM has 72.62% protein. This result can be attributed to the fact
that the lower viscosity of sacha inchi flour obtained by the wet method is due to the
sample;s particle size (sieving with a standard mesh size of 425 to 600 microns), as it
was not possible to reduce particle size due to its high fat content, and the resulting
granules from the liquefaction and subsequent drying processes were used.
Therefore, the endogenous lipids present in the taro flour form a type of protective
layer around the aggregate of carbohydrates and proteins, preventing water
absorption and the subsequent swelling of starch”.

3 Some parts were not correctly written, such as in page 10 where it was refer to
Table2 for oil concentration. While Table 2 was about antioxidant activity. Also,
statistical notation for Table 2, ABTS did not look right.
Regarding the suggested corrections, these have been properly adjusted, where Table 1 was
referenced and the scientific notation was modified.
Dear Dr Purwandari, Thank you very much for your thoughtful and constructive feedback on our manuscript. We appreciate the time and effort you dedicated to reviewing our work, as well as your insightful suggestions, which have greatly helped us improve our paper. Below, we address each of your comments in detail:

1.The paper was written carefully enough and data were analyzed thoroughly using
recent related publication. However, microbiological analysis did not seem relevant
to the work, and therefore it would be better deleted.
Regarding the microbiological analysis, it is important to note that this was conducted to
evaluate the initial state of the obtained flours concerning their microbiological
composition. Since these flours are intended for food consumption, it is essential to know
the level of microbial growth and to ensure that they fall within the permissible ranges
established by current regulations governing such products.

2. Some parts were not correctly written, such as in page 10 where it was refer to
Table2 for oil concentration. While Table 2 was about antioxidant activity. Also,
statistical notation for Table 2, ABTS did not look right.

With the purpose of addressing the question raised, the information was supplemented
using the correct term ;granulometry.; Additionally, the article mentions the following
textually “Comparing SIF-DM flour with SIF-WM, the defatting method produces a
flour with higher viscosity due to the interaction of water with proteins and starch,
considering that SIF-DM has 72.62% protein. This result can be attributed to the fact
that the lower viscosity of sacha inchi flour obtained by the wet method is due to the
sample;s particle size (sieving with a standard mesh size of 425 to 600 microns), as it
was not possible to reduce particle size due to its high fat content, and the resulting
granules from the liquefaction and subsequent drying processes were used.
Therefore, the endogenous lipids present in the taro flour form a type of protective
layer around the aggregate of carbohydrates and proteins, preventing water
absorption and the subsequent swelling of starch”.

3 Some parts were not correctly written, such as in page 10 where it was refer to
Table2 for oil concentration. While Table 2 was about antioxidant activity. Also,
statistical notation for Table 2, ABTS did not look right.
Regarding the suggested corrections, these have been properly adjusted, where Table 1 was
referenced and the scientific notation was modified.
Competing Interests: No competing interests were disclosed. Close
Report a concern
Respond or Comment

COMMENTS ON THIS REPORT

Author Response 22 Nov 2024

Karen Muñoz, Departamento de Agroindustria, Universidad del Cauca, Popayan, Colombia

22 Nov 2024

Author Response

Dear Dr Purwandari, Thank you very much for your thoughtful and constructive feedback on our manuscript. We appreciate the time and effort you dedicated to reviewing our work, as well ... Continue reading Dear Dr Purwandari, Thank you very much for your thoughtful and constructive feedback on our manuscript. We appreciate the time and effort you dedicated to reviewing our work, as well as your insightful suggestions, which have greatly helped us improve our paper. Below, we address each of your comments in detail:

1.The paper was written carefully enough and data were analyzed thoroughly using
recent related publication. However, microbiological analysis did not seem relevant
to the work, and therefore it would be better deleted.
Regarding the microbiological analysis, it is important to note that this was conducted to
evaluate the initial state of the obtained flours concerning their microbiological
composition. Since these flours are intended for food consumption, it is essential to know
the level of microbial growth and to ensure that they fall within the permissible ranges
established by current regulations governing such products.

2. Some parts were not correctly written, such as in page 10 where it was refer to
Table2 for oil concentration. While Table 2 was about antioxidant activity. Also,
statistical notation for Table 2, ABTS did not look right.

With the purpose of addressing the question raised, the information was supplemented
using the correct term ;granulometry.; Additionally, the article mentions the following
textually “Comparing SIF-DM flour with SIF-WM, the defatting method produces a
flour with higher viscosity due to the interaction of water with proteins and starch,
considering that SIF-DM has 72.62% protein. This result can be attributed to the fact
that the lower viscosity of sacha inchi flour obtained by the wet method is due to the
sample;s particle size (sieving with a standard mesh size of 425 to 600 microns), as it
was not possible to reduce particle size due to its high fat content, and the resulting
granules from the liquefaction and subsequent drying processes were used.
Therefore, the endogenous lipids present in the taro flour form a type of protective
layer around the aggregate of carbohydrates and proteins, preventing water
absorption and the subsequent swelling of starch”.

3 Some parts were not correctly written, such as in page 10 where it was refer to
Table2 for oil concentration. While Table 2 was about antioxidant activity. Also,
statistical notation for Table 2, ABTS did not look right.
Regarding the suggested corrections, these have been properly adjusted, where Table 1 was
referenced and the scientific notation was modified.
Dear Dr Purwandari, Thank you very much for your thoughtful and constructive feedback on our manuscript. We appreciate the time and effort you dedicated to reviewing our work, as well as your insightful suggestions, which have greatly helped us improve our paper. Below, we address each of your comments in detail:

1.The paper was written carefully enough and data were analyzed thoroughly using
recent related publication. However, microbiological analysis did not seem relevant
to the work, and therefore it would be better deleted.
Regarding the microbiological analysis, it is important to note that this was conducted to
evaluate the initial state of the obtained flours concerning their microbiological
composition. Since these flours are intended for food consumption, it is essential to know
the level of microbial growth and to ensure that they fall within the permissible ranges
established by current regulations governing such products.

2. Some parts were not correctly written, such as in page 10 where it was refer to
Table2 for oil concentration. While Table 2 was about antioxidant activity. Also,
statistical notation for Table 2, ABTS did not look right.

With the purpose of addressing the question raised, the information was supplemented
using the correct term ;granulometry.; Additionally, the article mentions the following
textually “Comparing SIF-DM flour with SIF-WM, the defatting method produces a
flour with higher viscosity due to the interaction of water with proteins and starch,
considering that SIF-DM has 72.62% protein. This result can be attributed to the fact
that the lower viscosity of sacha inchi flour obtained by the wet method is due to the
sample;s particle size (sieving with a standard mesh size of 425 to 600 microns), as it
was not possible to reduce particle size due to its high fat content, and the resulting
granules from the liquefaction and subsequent drying processes were used.
Therefore, the endogenous lipids present in the taro flour form a type of protective
layer around the aggregate of carbohydrates and proteins, preventing water
absorption and the subsequent swelling of starch”.

3 Some parts were not correctly written, such as in page 10 where it was refer to
Table2 for oil concentration. While Table 2 was about antioxidant activity. Also,
statistical notation for Table 2, ABTS did not look right.
Regarding the suggested corrections, these have been properly adjusted, where Table 1 was
referenced and the scientific notation was modified.
Competing Interests: No competing interests were disclosed. Close
Report a concern

Views

Reviewer Report 10 Sep 2024

Elisa Julianti, Department of Food Science Faculty of Agriculture, Universitas Sumatera Utara, Medan, North Sumatra, Indonesia

Approved with Reservations

https://doi.org/10.5256/f1000research.155702.r317400

The subject of this manuscript (“Characterization of sacha inchi (Plukenetia volubilis) and taro (Colocasia esculenta) flours with potential application in the preparation of both gluten-free and high protein foods”) falls inside the general scope of the F1000 Research. The study presents an interesting exploration of sacha inchi and taro flours as alternatives in gluten-free and high-protein food formulations. The choice of materials is commendable due to their nutritional benefits and potential applications. However, in my point of view, the paper does need some revision before indexing, as follows:
1. In the conclusion of abstract, Based on the characteristics obtained, what type of food is suitable for the application of sacha inchi and taro flour? For example, would it be bread, pasta, biscuits, or other types of products?
2. In the introduction, it should be justified what are the drawbacks of using sacha inchi and taro as single food ingredients? Why do you want to combine sacha inchi and taro flour as food ingredients?
3. In the mathods : The method does not describe the analytical procedure for sacha inchi almonds. Was any treatment applied to the seeds before analysis?
Is there a reason for using the ratio of SIF and TF?
4. In the discussion :
- It should be explained what causes the difference in protein content obtained through the wet method and the mechanical defatting method.
- Sacha inchi flour obtained through the wet method is flour derived from the dried, water-insoluble components. The water-soluble carbohydrate components are certainly not included in SIF. However, why is the carbohydrate content in sacha inchi almonds lower than in SIF-DM? Additionally, why is the carbohydrate content in SIF-DM lower, even though only the fat is removed during its production? Please explain.
- In SIF-DM, there is a release of phenolic compound due to involving pressure and temperature, but why is the FPP content in SIF-DM higher than in SIF-WM? Does the released FPP not get lost during the processing?
- It would be more meaningful if the researcher could show the starch content of each sample with different SIF and TF ratios.
- Based on Table 3, the aw for all three types of flour is <0.85, which should prevent the growth of microorganisms. How do you explain the presence of Salmonella and mold in your flour products?
5. In conclusion : What is the recommended ratio of SIF (to TF ?

Is the work clearly and accurately presented and does it cite the current literature?

Yes
Is the study design appropriate and is the work technically sound?

Partly
Are sufficient details of methods and analysis provided to allow replication by others?

Partly
If applicable, is the statistical analysis and its interpretation appropriate?

No
Are all the source data underlying the results available to ensure full reproducibility?

Yes
Are the conclusions drawn adequately supported by the results?

Yes

Competing Interests: No competing interests were disclosed.

Reviewer Expertise: Flour and Starch from Root and Tuber Crops as Alternatives to Wheat Flour

CITE

Report a concern

Author Response 19 Nov 2024

Karen Muñoz, Departamento de Agroindustria, Universidad del Cauca, Popayan, Colombia

19 Nov 2024

Author Response

Dear Dr Elisa,Thank you very much for your thoughtful and constructive feedback on our manuscript. We appreciate the time and effort you dedicated to reviewing our work, as well as ... Continue reading Dear Dr Elisa,Thank you very much for your thoughtful and constructive feedback on our manuscript. We appreciate the time and effort you dedicated to reviewing our work, as well as your insightful suggestions, which have greatly helped us improve our paper. Below, we address each of your comments in detail:

1. In conclusion of abstract, Based on the characteristics obtained, what type of food is suitable for the application of sacha inchi and taro flour? For example, would it be bread, pasta, biscuits, or other types of products?

In response to Question 1, the conclusion of the document has been adjusted to mention that, based on the obtained characteristics, “the analyzed flours represent a raw material with great potential for the development of products such as protein snacks due to the protein contribution of sacha inchi, good expansion due to taro starch, and being naturally gluten-free for individuals with celiac disease”.

2. In the introduction, it should be justified what are the drawbacks of using sacha inchi and taro as single food ingredients? Why do you want to combine sacha inchi and taro flour as food ingredients?
In response to Question 2, the introduction has been supplemented with information about some drawbacks or limitations of both sacha inchi and taro. “Sacha inchi can be unstable due to its high concentration of unsaturated fatty acids, making it sensitive to oxidation. Additionally, sacha inchi has limiting amino acids, such as lysine and tryptophan, which may cause low digestibility. Likewise, this seed contains antinutritional factors that limit its use as a raw material in the development of new products. On the other hand, taro contains antinutrients such as oxalates, which can restrict its fresh consumption due to the irritation they cause in the throat and mouth. However, the extrusion process significantly reduces these issues while simultaneously enhancing nutritional properties, such as increased protein digestibility. Based on this, combining sacha inchi and taro is an excellent option to complement their nutritional properties, increasing nutritional value and improving the stability and texture of the final product”.

3. In the methods : The method does not describe the analytical procedure for sacha inchi almonds. Was any treatment applied to the seeds before analysis? Is there a reason for using the ratio of SIF and TF?In relation to Question 3, in the section on methods used to obtain sacha inchi flour, additional information is provided about the process followed prior to the proximal characterization of the sacha inchi kernel. It is explained verbatim that “the sacha inchi seeds were received as white kernels, meaning without shell or husk. For the compositional analysis of the kernel, they were manually ground with a mortar to reduce their size, thus facilitating the relevant tests”. Furthermore, the reason for using the SIF and TF ratio lies in their high nutritional value (carbohydrates and protein) and the fact that both raw materials have had limited scientific and industrial advancement, presenting a significant opportunity to leverage their potential.

4. 4. In the discussion :
- It should be explained what causes the difference in protein content obtained through
the wet method and the mechanical defatting method.

To address this question, additional information has been included to explain the cause of
differences in protein content obtained through the wet milling method compared to the mechanical defatting method. The document states verbatim: “It is important to note that the protein content obtained through the wet milling method is lower compared to the mechanical defatting method. This is because, during the wet milling process, a milky suspension is generated, which is then filtered to separate the insoluble extract (cake) from the water-soluble extract (slurry). During filtration, part of the protein may remain in the soluble extract, which explains the lower protein concentration in the cake. In this study, the proximate analysis was performed only on the insoluble extract”.

- Sacha inchi flour obtained through the wet method is flour derived from the dried,
water-insoluble components. The water-soluble carbohydrate components are certainly not
included in SIF. However, why is the carbohydrate content in sacha inchi almonds lower
than in SIF-DM? Additionally, why is the carbohydrate content in SIF-DM lower, even
though only the fat is removed during its production? Please explain.

Regarding the question, why is the carbohydrate content in sacha inchi kernels lower than
in SIF-DM? Table 1 shows that the kernel contains 6.67% carbohydrates, while SIF-DM
showed 3.04% carbohydrates. In relation to the question of why the carbohydrate content in
SIF-DM is lower, despite the fact that only fat is removed during its production: as shown
in the following image, it is important to mention that during the oil extraction process, not
only fat is removed but also some solids such as proteins, minerals, and some
carbohydrates. These components may become trapped in the oil or be removed with the
byproduct generated during extraction. Additionally, it is likely that some soluble
carbohydrates dissolve or are removed along with the oil during mechanical extraction.

- In SIF-DM, there is a release of phenolic compounds due to involving pressure and
temperature, but why is the FPP content in SIF-DM higher than in SIF-WM? Does the
released FPP not get lost during the processing?

Regarding the question of why the free total phenol content (FPP) in SIF-DM is higher than
in SIF-WM, it is important to consider the differences in the extraction processes. During
the wet milling process (SIF-WM), phenolic compounds, which are water-soluble, can
migrate into the liquid solvent during the maceration stage, resulting in a significant loss of
these bioactive compounds in the solvent and thus decreasing the concentration of phenols
in the final flour. In contrast, the mechanical defatting process (SIF-DM), a more physical
method that does not involve liquid solvents, results in much lower phenolic compound
loss. Without a significant aqueous phase, the phenols remain largely within the solid
matrix of the flour.

- It would be more meaningful if the researcher could show the starch content of each
sample with different SIF and TF ratios.

To complement this information in the article, the following text has been added detailing the
carbohydrate percentage and protein content in each mixture with different inclusion levels. “The study design was based on the type of flour and its inclusion levels in the mixture, taking protein content and carbohydrate percentage as the most relevant factors in each treatment. The following results were obtained: the 25% SIF-DM:75% TF mixture had 22.69% protein and 64.83%
carbohydrates; the 50% SIF-DM:50% TF mixture showed 39.34% protein and 44.23%
carbohydrates; and the 75% SIF-DM:25% TF mixture yielded 55.98% protein and 23.64%
carbohydrates. Meanwhile, the mixtures with SIF-WM presented the following values: 25% SIF-
WM:75% TF with 12.42% protein and 73.47% carbohydrates; 50% SIF-WM:50% TF with 18.80% protein and 61.52% carbohydrates; and finally, the 75% SIF-WM:25% TF mixture with 25.17% protein and 49.56% carbohydrates”.

- Based on Table 3, the aw for all three types of flour is <0.85, which should prevent the
growth of microorganisms. How do you explain the presence of Salmonella and mold in
your flour products?

As mentioned in Table 3, the analyzed flours exhibit low water activity, with values below 0.85.
This level of water activity is not suitable for the growth of most microorganisms, as the available water is insufficient to sustain basic metabolic processes. However, while low water activity inhibits the active proliferation of microorganisms, it does not guarantee the total absence of pathogens. Spores of certain microorganisms and bacteria, such as Salmonella, can survive under adverse conditions, remaining viable for extended periods. Additionally, many pathogens possess persistence mechanisms that allow them to survive in hostile environments. In this case, Salmonella can form protective structures called biofilms, which enable microorganisms to adhere to surfaces
and resist disinfection treatments. Moreover, these biofilms not only act as physical barriers but also increase the tolerance of microorganisms to physical processes such as drying, protecting bacterial cells from environmental stress.
5. In conclusion : What is the recommended ratio of SIF to TF ?
To address this question, the conclusions state that through the viscoelasticity analysis (temperature sweep), it was determined that the optimal inclusion level of defatted sacha inchi is between 25% and 50%, which allows for the formation of a stable and consistent gel. Complementing this conclusion, the 25% SIF-DM:75% TF mixture has 22.69% protein and 64.83% carbohydrates, while the 50% SIF-DM:50% TF mixture shows 39.34% protein and 44.23% carbohydrates. These values are relevant for the development of products such as extruded snacks, where the protein contribution from sacha inchi is crucial, and the expansion capability provided by taro starch allows both ingredients to complement each other effectively.
Dear Dr Elisa,Thank you very much for your thoughtful and constructive feedback on our manuscript. We appreciate the time and effort you dedicated to reviewing our work, as well as your insightful suggestions, which have greatly helped us improve our paper. Below, we address each of your comments in detail:

1. In conclusion of abstract, Based on the characteristics obtained, what type of food is suitable for the application of sacha inchi and taro flour? For example, would it be bread, pasta, biscuits, or other types of products?

In response to Question 1, the conclusion of the document has been adjusted to mention that, based on the obtained characteristics, “the analyzed flours represent a raw material with great potential for the development of products such as protein snacks due to the protein contribution of sacha inchi, good expansion due to taro starch, and being naturally gluten-free for individuals with celiac disease”.

2. In the introduction, it should be justified what are the drawbacks of using sacha inchi and taro as single food ingredients? Why do you want to combine sacha inchi and taro flour as food ingredients?
In response to Question 2, the introduction has been supplemented with information about some drawbacks or limitations of both sacha inchi and taro. “Sacha inchi can be unstable due to its high concentration of unsaturated fatty acids, making it sensitive to oxidation. Additionally, sacha inchi has limiting amino acids, such as lysine and tryptophan, which may cause low digestibility. Likewise, this seed contains antinutritional factors that limit its use as a raw material in the development of new products. On the other hand, taro contains antinutrients such as oxalates, which can restrict its fresh consumption due to the irritation they cause in the throat and mouth. However, the extrusion process significantly reduces these issues while simultaneously enhancing nutritional properties, such as increased protein digestibility. Based on this, combining sacha inchi and taro is an excellent option to complement their nutritional properties, increasing nutritional value and improving the stability and texture of the final product”.

3. In the methods : The method does not describe the analytical procedure for sacha inchi almonds. Was any treatment applied to the seeds before analysis? Is there a reason for using the ratio of SIF and TF?In relation to Question 3, in the section on methods used to obtain sacha inchi flour, additional information is provided about the process followed prior to the proximal characterization of the sacha inchi kernel. It is explained verbatim that “the sacha inchi seeds were received as white kernels, meaning without shell or husk. For the compositional analysis of the kernel, they were manually ground with a mortar to reduce their size, thus facilitating the relevant tests”. Furthermore, the reason for using the SIF and TF ratio lies in their high nutritional value (carbohydrates and protein) and the fact that both raw materials have had limited scientific and industrial advancement, presenting a significant opportunity to leverage their potential.

4. 4. In the discussion :
- It should be explained what causes the difference in protein content obtained through
the wet method and the mechanical defatting method.

To address this question, additional information has been included to explain the cause of
differences in protein content obtained through the wet milling method compared to the mechanical defatting method. The document states verbatim: “It is important to note that the protein content obtained through the wet milling method is lower compared to the mechanical defatting method. This is because, during the wet milling process, a milky suspension is generated, which is then filtered to separate the insoluble extract (cake) from the water-soluble extract (slurry). During filtration, part of the protein may remain in the soluble extract, which explains the lower protein concentration in the cake. In this study, the proximate analysis was performed only on the insoluble extract”.

- Sacha inchi flour obtained through the wet method is flour derived from the dried,
water-insoluble components. The water-soluble carbohydrate components are certainly not
included in SIF. However, why is the carbohydrate content in sacha inchi almonds lower
than in SIF-DM? Additionally, why is the carbohydrate content in SIF-DM lower, even
though only the fat is removed during its production? Please explain.

Regarding the question, why is the carbohydrate content in sacha inchi kernels lower than
in SIF-DM? Table 1 shows that the kernel contains 6.67% carbohydrates, while SIF-DM
showed 3.04% carbohydrates. In relation to the question of why the carbohydrate content in
SIF-DM is lower, despite the fact that only fat is removed during its production: as shown
in the following image, it is important to mention that during the oil extraction process, not
only fat is removed but also some solids such as proteins, minerals, and some
carbohydrates. These components may become trapped in the oil or be removed with the
byproduct generated during extraction. Additionally, it is likely that some soluble
carbohydrates dissolve or are removed along with the oil during mechanical extraction.

- In SIF-DM, there is a release of phenolic compounds due to involving pressure and
temperature, but why is the FPP content in SIF-DM higher than in SIF-WM? Does the
released FPP not get lost during the processing?

Regarding the question of why the free total phenol content (FPP) in SIF-DM is higher than
in SIF-WM, it is important to consider the differences in the extraction processes. During
the wet milling process (SIF-WM), phenolic compounds, which are water-soluble, can
migrate into the liquid solvent during the maceration stage, resulting in a significant loss of
these bioactive compounds in the solvent and thus decreasing the concentration of phenols
in the final flour. In contrast, the mechanical defatting process (SIF-DM), a more physical
method that does not involve liquid solvents, results in much lower phenolic compound
loss. Without a significant aqueous phase, the phenols remain largely within the solid
matrix of the flour.

- It would be more meaningful if the researcher could show the starch content of each
sample with different SIF and TF ratios.

To complement this information in the article, the following text has been added detailing the
carbohydrate percentage and protein content in each mixture with different inclusion levels. “The study design was based on the type of flour and its inclusion levels in the mixture, taking protein content and carbohydrate percentage as the most relevant factors in each treatment. The following results were obtained: the 25% SIF-DM:75% TF mixture had 22.69% protein and 64.83%
carbohydrates; the 50% SIF-DM:50% TF mixture showed 39.34% protein and 44.23%
carbohydrates; and the 75% SIF-DM:25% TF mixture yielded 55.98% protein and 23.64%
carbohydrates. Meanwhile, the mixtures with SIF-WM presented the following values: 25% SIF-
WM:75% TF with 12.42% protein and 73.47% carbohydrates; 50% SIF-WM:50% TF with 18.80% protein and 61.52% carbohydrates; and finally, the 75% SIF-WM:25% TF mixture with 25.17% protein and 49.56% carbohydrates”.

- Based on Table 3, the aw for all three types of flour is <0.85, which should prevent the
growth of microorganisms. How do you explain the presence of Salmonella and mold in
your flour products?

As mentioned in Table 3, the analyzed flours exhibit low water activity, with values below 0.85.
This level of water activity is not suitable for the growth of most microorganisms, as the available water is insufficient to sustain basic metabolic processes. However, while low water activity inhibits the active proliferation of microorganisms, it does not guarantee the total absence of pathogens. Spores of certain microorganisms and bacteria, such as Salmonella, can survive under adverse conditions, remaining viable for extended periods. Additionally, many pathogens possess persistence mechanisms that allow them to survive in hostile environments. In this case, Salmonella can form protective structures called biofilms, which enable microorganisms to adhere to surfaces
and resist disinfection treatments. Moreover, these biofilms not only act as physical barriers but also increase the tolerance of microorganisms to physical processes such as drying, protecting bacterial cells from environmental stress.
5. In conclusion : What is the recommended ratio of SIF to TF ?
To address this question, the conclusions state that through the viscoelasticity analysis (temperature sweep), it was determined that the optimal inclusion level of defatted sacha inchi is between 25% and 50%, which allows for the formation of a stable and consistent gel. Complementing this conclusion, the 25% SIF-DM:75% TF mixture has 22.69% protein and 64.83% carbohydrates, while the 50% SIF-DM:50% TF mixture shows 39.34% protein and 44.23% carbohydrates. These values are relevant for the development of products such as extruded snacks, where the protein contribution from sacha inchi is crucial, and the expansion capability provided by taro starch allows both ingredients to complement each other effectively.
Competing Interests: No competing interests were disclosed. Close
Report a concern
Respond or Comment

COMMENTS ON THIS REPORT

Author Response 19 Nov 2024

Karen Muñoz, Departamento de Agroindustria, Universidad del Cauca, Popayan, Colombia

19 Nov 2024

Author Response

Dear Dr Elisa,Thank you very much for your thoughtful and constructive feedback on our manuscript. We appreciate the time and effort you dedicated to reviewing our work, as well as ... Continue reading Dear Dr Elisa,Thank you very much for your thoughtful and constructive feedback on our manuscript. We appreciate the time and effort you dedicated to reviewing our work, as well as your insightful suggestions, which have greatly helped us improve our paper. Below, we address each of your comments in detail:

1. In conclusion of abstract, Based on the characteristics obtained, what type of food is suitable for the application of sacha inchi and taro flour? For example, would it be bread, pasta, biscuits, or other types of products?

In response to Question 1, the conclusion of the document has been adjusted to mention that, based on the obtained characteristics, “the analyzed flours represent a raw material with great potential for the development of products such as protein snacks due to the protein contribution of sacha inchi, good expansion due to taro starch, and being naturally gluten-free for individuals with celiac disease”.

2. In the introduction, it should be justified what are the drawbacks of using sacha inchi and taro as single food ingredients? Why do you want to combine sacha inchi and taro flour as food ingredients?
In response to Question 2, the introduction has been supplemented with information about some drawbacks or limitations of both sacha inchi and taro. “Sacha inchi can be unstable due to its high concentration of unsaturated fatty acids, making it sensitive to oxidation. Additionally, sacha inchi has limiting amino acids, such as lysine and tryptophan, which may cause low digestibility. Likewise, this seed contains antinutritional factors that limit its use as a raw material in the development of new products. On the other hand, taro contains antinutrients such as oxalates, which can restrict its fresh consumption due to the irritation they cause in the throat and mouth. However, the extrusion process significantly reduces these issues while simultaneously enhancing nutritional properties, such as increased protein digestibility. Based on this, combining sacha inchi and taro is an excellent option to complement their nutritional properties, increasing nutritional value and improving the stability and texture of the final product”.

3. In the methods : The method does not describe the analytical procedure for sacha inchi almonds. Was any treatment applied to the seeds before analysis? Is there a reason for using the ratio of SIF and TF?In relation to Question 3, in the section on methods used to obtain sacha inchi flour, additional information is provided about the process followed prior to the proximal characterization of the sacha inchi kernel. It is explained verbatim that “the sacha inchi seeds were received as white kernels, meaning without shell or husk. For the compositional analysis of the kernel, they were manually ground with a mortar to reduce their size, thus facilitating the relevant tests”. Furthermore, the reason for using the SIF and TF ratio lies in their high nutritional value (carbohydrates and protein) and the fact that both raw materials have had limited scientific and industrial advancement, presenting a significant opportunity to leverage their potential.

4. 4. In the discussion :
- It should be explained what causes the difference in protein content obtained through
the wet method and the mechanical defatting method.

To address this question, additional information has been included to explain the cause of
differences in protein content obtained through the wet milling method compared to the mechanical defatting method. The document states verbatim: “It is important to note that the protein content obtained through the wet milling method is lower compared to the mechanical defatting method. This is because, during the wet milling process, a milky suspension is generated, which is then filtered to separate the insoluble extract (cake) from the water-soluble extract (slurry). During filtration, part of the protein may remain in the soluble extract, which explains the lower protein concentration in the cake. In this study, the proximate analysis was performed only on the insoluble extract”.

- Sacha inchi flour obtained through the wet method is flour derived from the dried,
water-insoluble components. The water-soluble carbohydrate components are certainly not
included in SIF. However, why is the carbohydrate content in sacha inchi almonds lower
than in SIF-DM? Additionally, why is the carbohydrate content in SIF-DM lower, even
though only the fat is removed during its production? Please explain.

Regarding the question, why is the carbohydrate content in sacha inchi kernels lower than
in SIF-DM? Table 1 shows that the kernel contains 6.67% carbohydrates, while SIF-DM
showed 3.04% carbohydrates. In relation to the question of why the carbohydrate content in
SIF-DM is lower, despite the fact that only fat is removed during its production: as shown
in the following image, it is important to mention that during the oil extraction process, not
only fat is removed but also some solids such as proteins, minerals, and some
carbohydrates. These components may become trapped in the oil or be removed with the
byproduct generated during extraction. Additionally, it is likely that some soluble
carbohydrates dissolve or are removed along with the oil during mechanical extraction.

- In SIF-DM, there is a release of phenolic compounds due to involving pressure and
temperature, but why is the FPP content in SIF-DM higher than in SIF-WM? Does the
released FPP not get lost during the processing?

Regarding the question of why the free total phenol content (FPP) in SIF-DM is higher than
in SIF-WM, it is important to consider the differences in the extraction processes. During
the wet milling process (SIF-WM), phenolic compounds, which are water-soluble, can
migrate into the liquid solvent during the maceration stage, resulting in a significant loss of
these bioactive compounds in the solvent and thus decreasing the concentration of phenols
in the final flour. In contrast, the mechanical defatting process (SIF-DM), a more physical
method that does not involve liquid solvents, results in much lower phenolic compound
loss. Without a significant aqueous phase, the phenols remain largely within the solid
matrix of the flour.

- It would be more meaningful if the researcher could show the starch content of each
sample with different SIF and TF ratios.

To complement this information in the article, the following text has been added detailing the
carbohydrate percentage and protein content in each mixture with different inclusion levels. “The study design was based on the type of flour and its inclusion levels in the mixture, taking protein content and carbohydrate percentage as the most relevant factors in each treatment. The following results were obtained: the 25% SIF-DM:75% TF mixture had 22.69% protein and 64.83%
carbohydrates; the 50% SIF-DM:50% TF mixture showed 39.34% protein and 44.23%
carbohydrates; and the 75% SIF-DM:25% TF mixture yielded 55.98% protein and 23.64%
carbohydrates. Meanwhile, the mixtures with SIF-WM presented the following values: 25% SIF-
WM:75% TF with 12.42% protein and 73.47% carbohydrates; 50% SIF-WM:50% TF with 18.80% protein and 61.52% carbohydrates; and finally, the 75% SIF-WM:25% TF mixture with 25.17% protein and 49.56% carbohydrates”.

- Based on Table 3, the aw for all three types of flour is <0.85, which should prevent the
growth of microorganisms. How do you explain the presence of Salmonella and mold in
your flour products?

As mentioned in Table 3, the analyzed flours exhibit low water activity, with values below 0.85.
This level of water activity is not suitable for the growth of most microorganisms, as the available water is insufficient to sustain basic metabolic processes. However, while low water activity inhibits the active proliferation of microorganisms, it does not guarantee the total absence of pathogens. Spores of certain microorganisms and bacteria, such as Salmonella, can survive under adverse conditions, remaining viable for extended periods. Additionally, many pathogens possess persistence mechanisms that allow them to survive in hostile environments. In this case, Salmonella can form protective structures called biofilms, which enable microorganisms to adhere to surfaces
and resist disinfection treatments. Moreover, these biofilms not only act as physical barriers but also increase the tolerance of microorganisms to physical processes such as drying, protecting bacterial cells from environmental stress.
5. In conclusion : What is the recommended ratio of SIF to TF ?
To address this question, the conclusions state that through the viscoelasticity analysis (temperature sweep), it was determined that the optimal inclusion level of defatted sacha inchi is between 25% and 50%, which allows for the formation of a stable and consistent gel. Complementing this conclusion, the 25% SIF-DM:75% TF mixture has 22.69% protein and 64.83% carbohydrates, while the 50% SIF-DM:50% TF mixture shows 39.34% protein and 44.23% carbohydrates. These values are relevant for the development of products such as extruded snacks, where the protein contribution from sacha inchi is crucial, and the expansion capability provided by taro starch allows both ingredients to complement each other effectively.
Dear Dr Elisa,Thank you very much for your thoughtful and constructive feedback on our manuscript. We appreciate the time and effort you dedicated to reviewing our work, as well as your insightful suggestions, which have greatly helped us improve our paper. Below, we address each of your comments in detail:

1. In conclusion of abstract, Based on the characteristics obtained, what type of food is suitable for the application of sacha inchi and taro flour? For example, would it be bread, pasta, biscuits, or other types of products?

In response to Question 1, the conclusion of the document has been adjusted to mention that, based on the obtained characteristics, “the analyzed flours represent a raw material with great potential for the development of products such as protein snacks due to the protein contribution of sacha inchi, good expansion due to taro starch, and being naturally gluten-free for individuals with celiac disease”.

2. In the introduction, it should be justified what are the drawbacks of using sacha inchi and taro as single food ingredients? Why do you want to combine sacha inchi and taro flour as food ingredients?
In response to Question 2, the introduction has been supplemented with information about some drawbacks or limitations of both sacha inchi and taro. “Sacha inchi can be unstable due to its high concentration of unsaturated fatty acids, making it sensitive to oxidation. Additionally, sacha inchi has limiting amino acids, such as lysine and tryptophan, which may cause low digestibility. Likewise, this seed contains antinutritional factors that limit its use as a raw material in the development of new products. On the other hand, taro contains antinutrients such as oxalates, which can restrict its fresh consumption due to the irritation they cause in the throat and mouth. However, the extrusion process significantly reduces these issues while simultaneously enhancing nutritional properties, such as increased protein digestibility. Based on this, combining sacha inchi and taro is an excellent option to complement their nutritional properties, increasing nutritional value and improving the stability and texture of the final product”.

3. In the methods : The method does not describe the analytical procedure for sacha inchi almonds. Was any treatment applied to the seeds before analysis? Is there a reason for using the ratio of SIF and TF?In relation to Question 3, in the section on methods used to obtain sacha inchi flour, additional information is provided about the process followed prior to the proximal characterization of the sacha inchi kernel. It is explained verbatim that “the sacha inchi seeds were received as white kernels, meaning without shell or husk. For the compositional analysis of the kernel, they were manually ground with a mortar to reduce their size, thus facilitating the relevant tests”. Furthermore, the reason for using the SIF and TF ratio lies in their high nutritional value (carbohydrates and protein) and the fact that both raw materials have had limited scientific and industrial advancement, presenting a significant opportunity to leverage their potential.

4. 4. In the discussion :
- It should be explained what causes the difference in protein content obtained through
the wet method and the mechanical defatting method.

To address this question, additional information has been included to explain the cause of
differences in protein content obtained through the wet milling method compared to the mechanical defatting method. The document states verbatim: “It is important to note that the protein content obtained through the wet milling method is lower compared to the mechanical defatting method. This is because, during the wet milling process, a milky suspension is generated, which is then filtered to separate the insoluble extract (cake) from the water-soluble extract (slurry). During filtration, part of the protein may remain in the soluble extract, which explains the lower protein concentration in the cake. In this study, the proximate analysis was performed only on the insoluble extract”.

- Sacha inchi flour obtained through the wet method is flour derived from the dried,
water-insoluble components. The water-soluble carbohydrate components are certainly not
included in SIF. However, why is the carbohydrate content in sacha inchi almonds lower
than in SIF-DM? Additionally, why is the carbohydrate content in SIF-DM lower, even
though only the fat is removed during its production? Please explain.

Regarding the question, why is the carbohydrate content in sacha inchi kernels lower than
in SIF-DM? Table 1 shows that the kernel contains 6.67% carbohydrates, while SIF-DM
showed 3.04% carbohydrates. In relation to the question of why the carbohydrate content in
SIF-DM is lower, despite the fact that only fat is removed during its production: as shown
in the following image, it is important to mention that during the oil extraction process, not
only fat is removed but also some solids such as proteins, minerals, and some
carbohydrates. These components may become trapped in the oil or be removed with the
byproduct generated during extraction. Additionally, it is likely that some soluble
carbohydrates dissolve or are removed along with the oil during mechanical extraction.

- In SIF-DM, there is a release of phenolic compounds due to involving pressure and
temperature, but why is the FPP content in SIF-DM higher than in SIF-WM? Does the
released FPP not get lost during the processing?

Regarding the question of why the free total phenol content (FPP) in SIF-DM is higher than
in SIF-WM, it is important to consider the differences in the extraction processes. During
the wet milling process (SIF-WM), phenolic compounds, which are water-soluble, can
migrate into the liquid solvent during the maceration stage, resulting in a significant loss of
these bioactive compounds in the solvent and thus decreasing the concentration of phenols
in the final flour. In contrast, the mechanical defatting process (SIF-DM), a more physical
method that does not involve liquid solvents, results in much lower phenolic compound
loss. Without a significant aqueous phase, the phenols remain largely within the solid
matrix of the flour.

- It would be more meaningful if the researcher could show the starch content of each
sample with different SIF and TF ratios.

To complement this information in the article, the following text has been added detailing the
carbohydrate percentage and protein content in each mixture with different inclusion levels. “The study design was based on the type of flour and its inclusion levels in the mixture, taking protein content and carbohydrate percentage as the most relevant factors in each treatment. The following results were obtained: the 25% SIF-DM:75% TF mixture had 22.69% protein and 64.83%
carbohydrates; the 50% SIF-DM:50% TF mixture showed 39.34% protein and 44.23%
carbohydrates; and the 75% SIF-DM:25% TF mixture yielded 55.98% protein and 23.64%
carbohydrates. Meanwhile, the mixtures with SIF-WM presented the following values: 25% SIF-
WM:75% TF with 12.42% protein and 73.47% carbohydrates; 50% SIF-WM:50% TF with 18.80% protein and 61.52% carbohydrates; and finally, the 75% SIF-WM:25% TF mixture with 25.17% protein and 49.56% carbohydrates”.

- Based on Table 3, the aw for all three types of flour is <0.85, which should prevent the
growth of microorganisms. How do you explain the presence of Salmonella and mold in
your flour products?

As mentioned in Table 3, the analyzed flours exhibit low water activity, with values below 0.85.
This level of water activity is not suitable for the growth of most microorganisms, as the available water is insufficient to sustain basic metabolic processes. However, while low water activity inhibits the active proliferation of microorganisms, it does not guarantee the total absence of pathogens. Spores of certain microorganisms and bacteria, such as Salmonella, can survive under adverse conditions, remaining viable for extended periods. Additionally, many pathogens possess persistence mechanisms that allow them to survive in hostile environments. In this case, Salmonella can form protective structures called biofilms, which enable microorganisms to adhere to surfaces
and resist disinfection treatments. Moreover, these biofilms not only act as physical barriers but also increase the tolerance of microorganisms to physical processes such as drying, protecting bacterial cells from environmental stress.
5. In conclusion : What is the recommended ratio of SIF to TF ?
To address this question, the conclusions state that through the viscoelasticity analysis (temperature sweep), it was determined that the optimal inclusion level of defatted sacha inchi is between 25% and 50%, which allows for the formation of a stable and consistent gel. Complementing this conclusion, the 25% SIF-DM:75% TF mixture has 22.69% protein and 64.83% carbohydrates, while the 50% SIF-DM:50% TF mixture shows 39.34% protein and 44.23% carbohydrates. These values are relevant for the development of products such as extruded snacks, where the protein contribution from sacha inchi is crucial, and the expansion capability provided by taro starch allows both ingredients to complement each other effectively.
Competing Interests: No competing interests were disclosed. Close
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Version 1

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PUBLISHED 06 Apr 2023

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Reviewer Report 04 Sep 2023

Diego Salazar, Facultad de Ciencia e Ingeniería en Alimentos, Universidad Tecnica de Ambato, Ambato, Tungurahua, Ecuador; Universidad Complutense de Madrid, Madrid, Community of Madrid, Spain

Not Approved

https://doi.org/10.5256/f1000research.143149.r175167

The work presented is interested in using unconventional sources, their characterization, and their nutritional components. However, the work requires management of scientific writing, the information presented needs to be revised, and the correct use of units of measurement is missed, for example, %, degrees Celsius, among others. On the other hand, the methodology needs to be more precise and allow a straightforward reading of what has been developed. Finally, the results must be rewritten, and consider that critical analysis and discussion of the results must be carried out, not only using the data from the tables and rewriting them.

Is the work clearly and accurately presented and does it cite the current literature?

No
Is the study design appropriate and is the work technically sound?

Partly
Are sufficient details of methods and analysis provided to allow replication by others?

Partly
If applicable, is the statistical analysis and its interpretation appropriate?

No
Are all the source data underlying the results available to ensure full reproducibility?

No
Are the conclusions drawn adequately supported by the results?

Partly

Competing Interests: No competing interests were disclosed.

Reviewer Expertise: Food Science and Technology. Research in new sources of foods, andean crops, legumes, tubers.

I confirm that I have read this submission and believe that I have an appropriate level of expertise to state that I do not consider it to be of an acceptable scientific standard, for reasons outlined above.

CITE

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Author Response 09 Oct 2023

Karen Muñoz, Departamento de Agroindustria, Universidad del Cauca, Popayan, Colombia

09 Oct 2023

Author Response

The units are adjusted and the methodology is clarified.

We kindly ask you to specify in which paragraph it is necessary to adjust the analysis, since in each test ... Continue reading The units are adjusted and the methodology is clarified.

We kindly ask you to specify in which paragraph it is necessary to adjust the analysis, since in each test a comparative and statistical analysis of the results is performed.
The units are adjusted and the methodology is clarified.

We kindly ask you to specify in which paragraph it is necessary to adjust the analysis, since in each test a comparative and statistical analysis of the results is performed.
Competing Interests: No competing interests were disclosed. Close
Report a concern
Author Response 13 Apr 2024

Karen Muñoz, Departamento de Agroindustria, Universidad del Cauca, Popayan, Colombia

13 Apr 2024

Author Response

Thank you for your valuable comments.

I would like to continue the process of evaluation and correction of the article, for this I ask for new corrections, in order ... Continue reading Thank you for your valuable comments.

I would like to continue the process of evaluation and correction of the article, for this I ask for new corrections, in order to be able to make them.

Best regards
Thank you for your valuable comments.

I would like to continue the process of evaluation and correction of the article, for this I ask for new corrections, in order to be able to make them.

Best regards
Competing Interests: No competing interests were disclosed. Close
Report a concern
Respond or Comment

COMMENTS ON THIS REPORT

Author Response 09 Oct 2023

Karen Muñoz, Departamento de Agroindustria, Universidad del Cauca, Popayan, Colombia

09 Oct 2023

Author Response

The units are adjusted and the methodology is clarified.

We kindly ask you to specify in which paragraph it is necessary to adjust the analysis, since in each test ... Continue reading The units are adjusted and the methodology is clarified.

We kindly ask you to specify in which paragraph it is necessary to adjust the analysis, since in each test a comparative and statistical analysis of the results is performed.
The units are adjusted and the methodology is clarified.

We kindly ask you to specify in which paragraph it is necessary to adjust the analysis, since in each test a comparative and statistical analysis of the results is performed.
Competing Interests: No competing interests were disclosed. Close
Report a concern
Author Response 13 Apr 2024

Karen Muñoz, Departamento de Agroindustria, Universidad del Cauca, Popayan, Colombia

13 Apr 2024

Author Response

Thank you for your valuable comments.

I would like to continue the process of evaluation and correction of the article, for this I ask for new corrections, in order ... Continue reading Thank you for your valuable comments.

I would like to continue the process of evaluation and correction of the article, for this I ask for new corrections, in order to be able to make them.

Best regards
Thank you for your valuable comments.

I would like to continue the process of evaluation and correction of the article, for this I ask for new corrections, in order to be able to make them.

Best regards
Competing Interests: No competing interests were disclosed. Close
Report a concern

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Reviewer Report 04 Sep 2023

Oluwaseun P Bamidele, University of Venda, Thohoyandou, Limpopo, South Africa

Not Approved

https://doi.org/10.5256/f1000research.143149.r195999

I worked on the PDF format of this manuscript and there is a fundamental error in the design of this study. The work would have been good if the authors work on compositing the two raw materials. There is no scientific contribution with way the study is presented. Although the Sacha inchi is new but not Taro.

The major challenge with the manuscript is the design. Compositing the root (Taro) and the Sacha at different ratios or percentages will make a difference in flour, which is rich in protein and other bioactive compounds.

Also, the manner in which the author reported the analyses is poor. Reporting on Sacha is not enough, using it in food preparation will be the best.

I recommend that the authors should proofread their manuscript. The English is not good enough.

Please find the annotated pdf with further comments here.

Is the work clearly and accurately presented and does it cite the current literature?

No
Is the study design appropriate and is the work technically sound?

No
Are sufficient details of methods and analysis provided to allow replication by others?

Yes
If applicable, is the statistical analysis and its interpretation appropriate?

No
Are all the source data underlying the results available to ensure full reproducibility?

Yes
Are the conclusions drawn adequately supported by the results?

No

Competing Interests: No competing interests were disclosed.

Reviewer Expertise: Food Chemistry, Food processing and preservation, Encapsulation of bioactive compounds, Functional foods, Gamma irradiation.

CITE

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Author Response 09 Oct 2023

Karen Muñoz, Departamento de Agroindustria, Universidad del Cauca, Popayan, Colombia

09 Oct 2023

Author Response

The corrections suggested in the pdf of the manuscript were adjusted.

A characterization of the two raw materials taro and sacha inchi is made.

This research was done ... Continue reading The corrections suggested in the pdf of the manuscript were adjusted.

A characterization of the two raw materials taro and sacha inchi is made.

This research was done in order to pave the way for new projects on the development of food based on vegetable matrices.

Mixtures with inclusion levels of 25, 50 and 75% of sacha inchi flour and taro 75, 50 and 25% were worked in order to evaluate the bioactive, rheological and proximal properties.

We kindly request to specify the paragraphs or sentences that should be adjusted to improve the analysis.

In addition, improvements were made in the English language.
The corrections suggested in the pdf of the manuscript were adjusted.

A characterization of the two raw materials taro and sacha inchi is made.

This research was done in order to pave the way for new projects on the development of food based on vegetable matrices.

Mixtures with inclusion levels of 25, 50 and 75% of sacha inchi flour and taro 75, 50 and 25% were worked in order to evaluate the bioactive, rheological and proximal properties.

We kindly request to specify the paragraphs or sentences that should be adjusted to improve the analysis.

In addition, improvements were made in the English language.
Competing Interests: No competing interests were disclosed. Close
Report a concern
Respond or Comment

COMMENTS ON THIS REPORT

Author Response 09 Oct 2023

Karen Muñoz, Departamento de Agroindustria, Universidad del Cauca, Popayan, Colombia

09 Oct 2023

Author Response

The corrections suggested in the pdf of the manuscript were adjusted.

A characterization of the two raw materials taro and sacha inchi is made.

This research was done ... Continue reading The corrections suggested in the pdf of the manuscript were adjusted.

A characterization of the two raw materials taro and sacha inchi is made.

This research was done in order to pave the way for new projects on the development of food based on vegetable matrices.

Mixtures with inclusion levels of 25, 50 and 75% of sacha inchi flour and taro 75, 50 and 25% were worked in order to evaluate the bioactive, rheological and proximal properties.

We kindly request to specify the paragraphs or sentences that should be adjusted to improve the analysis.

In addition, improvements were made in the English language.
The corrections suggested in the pdf of the manuscript were adjusted.

A characterization of the two raw materials taro and sacha inchi is made.

This research was done in order to pave the way for new projects on the development of food based on vegetable matrices.

Mixtures with inclusion levels of 25, 50 and 75% of sacha inchi flour and taro 75, 50 and 25% were worked in order to evaluate the bioactive, rheological and proximal properties.

We kindly request to specify the paragraphs or sentences that should be adjusted to improve the analysis.

In addition, improvements were made in the English language.
Competing Interests: No competing interests were disclosed. Close
Report a concern

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Version 4

VERSION 4 PUBLISHED 06 Apr 2023

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Oluwaseun P Bamidele, University of Venda, Thohoyandou, South Africa
Diego Salazar, Universidad Tecnica de Ambato, Ambato, Ecuador; Universidad Complutense de Madrid, Madrid, Spain
Elisa Julianti, Universitas Sumatera Utara, Medan, Indonesia
Umi Purwandari, University of Trunojoyo Madura, Madura, Indonesia
Lorena Sciarini, Universidad Nacional de Cordoba, Córdoba, Argentina

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02 Apr 2025 | for Version 4

Lorena Sciarini, Universidad Nacional de Cordoba, Córdoba, Cordoba, Argentina

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Approved With Reservations

The authors have made most of the requested revisions. However, some issues remain unresolved, and new ones have arisen, especially regarding SIF composition and rheological properties.
Pp 2. Abstract. 7.47 mg GAE/g corresponds to FPP content, not antioxidant activity.
Pp 5. Please, do not start sentences with a number. Check throughout the manuscript.
Pp 5. Protein determination. Which factor was used for protein calculation?
Pp 6. Please change “Fiber” to “Crude fiber.”
Table 1. “The lipids results were close, 9.84% for the SIF-DM and 8.87% for SIF-WM.” This result is not intuitive. While DM mainly extracts lipids, WM extracts hydrophilic compounds. Why do they have the same lipid content?
In general, the composition of SIF is striking, especially in terms of protein and lipid content (since carbohydrates are calculated by difference, their values are relative). The difference in protein concentration may be due to partial solubilization during the WM process. The difference from the almond is also noteworthy. Another remarkable aspect is the (small) difference in lipid content. I suggest the authors to review these values.
Pp 9. Please unify terminology throughout the text: dry method, defatted method, mechanical pressing.
Pp 9. “On the other hand, it is likely that carbohydrates are solubilized in the water used in the wet method, contributing to their higher content in SIF-WM.” This sentence is contradictory. Please revise.
Pp 10. Change “by 43” to “by Sánchez-Rivera 43.”
Figure 3B seems to present Trough viscosity values. Please check.
Pp 11. Change “and solubilizes amylose and amylopectin fractions” to “and amylose partially leaches out from granules” (the term “solubilizes” is not correct in this context).
Pp 11. “Reorganize into a structure similar to the original one” is not accurate. Starch gelatinization is an irreversible phenomenon. During cooling and storage, amylose and amylopectin do reorganize into partially crystalline structures, but these are completely different from the original starch granules.
Pp 11. “This behavior can be explained by the properties of starch, as during heating, low-molecular-weight amylose separates from the starch granule, causing it to collapse at a constant temperature as the amorphous portion is dispersed, resulting in a decrease in viscosity.” In general, breakdown positively correlates with peak viscosity: the more water the starch granule absorbs, the more fragile it becomes.
Pp 11. “According to the results, blends with higher TF content exhibited higher setback values, which is due to the higher amylopectin content in taro starch (66% amylopectin versus 34% amylose).” It is unclear what is being compared to what. The starch source is TF, which always has the same AM-AP ratio. Additionally, setback is related to amylose retrogradation, not amylopectin. Finally, in this study, the higher setback is likely related to the higher starch content.
Figure 4. Please clarify the image references (symbols) in the figure caption.
Figures 5A and 5H. The relatively high modulus values from the beginning of the test at room temperature are remarkable. Please comment.
Conclusion. “Mechanical defatting stood out as the most efficient method for obtaining high-protein flours from sacha inchi. However, this process was observed to reduce the preservation of bioactive compounds, decreasing antioxidant activity and total polyphenol content compared to the wet milling method.” This seems to be the opposite, as the FPP and ABTS values are higher in SIF-DM than in SIF-WM.

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

Gluten free bakery, starch properties, cereal properties

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Reviewer Report

24 Views

27 Dec 2024 | for Version 3

Lorena Sciarini, Universidad Nacional de Cordoba, Córdoba, Cordoba, Argentina

24 Views Cite this report Responses(1)

Approved With Reservations

Pp1. Abstract. “The ABTS radical method, which reacts with the phenolic compounds of the food matrix, using Trolox as a standard.” Please review the wording (the sentence is incomplete).

Pp 2. Does "Salmonella" correspond to a specific species? Please revise (it could also be Salmonella sp. or Salmonella spp.).

Pp2. It is suggested to change “According to Ref. 1” to “According to Wang et al.¹.” Consider applying this modification throughout the text.

Pp 2. “According to Ref. 1, by 2050, if governance allows for debate and equity in global food and if food demand is very high, driven by rapid population growth, a high intensity of land use will be required to meet the demand for food”. The wording is unclear. Please revise.

Pp 4. Please, replace “mNicroorganisms” with “microorganisms”

Pp4. “this, in order to grind the grain. Obtaining a milky and homogeneous” Please revise the punctuation (I understand the period is not appropriate).

Pp4. Deffating method. Was the extraction cake dried? How was it milled afterward? Was it sieved? Please complete the methodology.

Pp5. For the proximate composition, please specify which AOAC method was used in each case.

Pp6. Ref. 20 is incomplete in the References section. Please review all citations.

Pp 6. Polyphenols. “Finally, the supernatant mixture was made up to 20 mL using distilled water and the absorbance was measured in the spectrophotometer”. Which colorimetric technique was used? Folin-Ciocalteu? Please clarify.

Pp6. ABTS. “In a test tube 4mL of ABTS.* (2,20-Azino-bis(3-ethylbenzthiazoline-6-sulfonic acid), solution was added and to start the reaction 135 μL standard solution and vortexed for 5 seconds.” Please revise the wording.

Pp6. Flow profile. Which geometry was used?

Pp7. Fungi and yeast. “30°” with “30 °C”.

Pp8. Results and discussion. Table 1. The wet method of obtaining Sacha inchi flours results in the solubilization of hydrophilic compounds, such as proteins. This causes the concentration of other components to increase (particularly carbohydrates, according to the authors' findings). However, it is striking that the lipid concentration is so similar, given that the mechanical method specifically extracts the oil. What is the reason for this similarity?

Pp8. “As for carbohydrate content, SIF-DM showed a lower percentage of carbohydrates compared to other extraction methods, due to the fact that in the oil extraction process, not only fat is removed but also some solids such as proteins, minerals, and certain carbohydrates (see Figure 1B). These components may become trapped in the oil or be eliminated with the byproduct generated during extraction. Additionally, it is likely that some soluble carbohydrates dissolve or are removed along with the oil during mechanical extraction”. It is more likely that carbohydrates are solubilized in the water used in the wet method rather than in the oil, although some carryover may occur. It is worth exploring the fact that the wet method actually has a higher carbohydrate content because, proportionally, it has fewer proteins, leading to a concentration of carbohydrates (which are calculated by difference).

Pp9. Was the proximate composition of the flour blends measured or calculated?

Pp9. Effect of processing on FPP and the antioxidant activities. Please do not repeat in the text the values already presented in Tables/Figures. Verify this throughout the manuscript. Also, check, as "taro flour presents the highest polyphenol content of 7.50+/-1.73" does not match the value presented in Table 2.

Pp 10. “This difference can be explained because taro is a prime source of antioxidant” What is meant by "prime source"?

Pp10. “… bioactive compounds, which in sacha inchi are present particularly in polyunsaturated essential fatty acids…” ABTS is used to evaluate the radical scavenging capacity of hydrophilic compounds. In fact, the extraction was performed with polar solvents. I find the explanation contradictory.

Pp11. “to the post hoc Tukey analysis five different groups identified with letters from “a” to “e” are classified. In group “a”, the samples with the highest viscosity (25% SIF-DM:75% TF and 100% TF) with values of 3.08 Pa.s and 2.99 Pa.s respectively, and in group “e”, the samples with the lowest viscosity value (75% SIF-WM:25% TF and 100% SIF-WM) with values of 0.35 Pa.s and 0.64 Pa.s respectively.” This type of expression used throughout the manuscript makes the reading tedious and is repeated with what is presented in graphs/tables. Please simplify by focusing on the statistical differences presented between samples.

Pp12. In this page, the results are presented in a disorganized manner. It is suggested to first address Figure 2, and then Figure 3.

Pp12. “According to the above, as shown in Figure 3, the samples that presented a higher viscosity drop are the samples with higher TF content, this is because the crystalline structure (amylopectin) that makes up the granules is altered, when amylose and amylopectin are hydrated individually in excess water, both macromolecules tend to reorganize in a similar structure to the one they had initially.” The authors discuss the minimum viscosity (or trough), but I believe there is some confusion here, as this parameter is not associated with retrogradation but with gelatinization. The granules absorb water up to a maximum, and with shear at high temperature, they break, decreasing viscosity. Retrogradation occurs when the temperature decreases.

Pp12. “As shown in Figure 3, mixtures with higher inclusion of starch present in TF and less protein contributed by sacha inchi result in more free water for starch molecules to swell and solubilize presenting higher viscosity; on the contrary, at high proportions of protein its interaction with starch granules could stabilize the amylose leaching channel of starch granules, which resulted in lower swelling power and solubility.” It would be useful to consider, in the mixtures, the effect of dilution of the starch present in TF. On the other hand, proteins also develop viscosity, which could explain why TF+SIF-DM has higher viscosity than TF+SIF-WM.

Pp12. “until the amorphous part is solubilized, decreasing viscosity”. In general, when referring to starch in water, the term "dispersion" is used, not "solubilization."

Pp12. Flow profile analysis. Please, unify: “flowability index” or “flow index”.

Pp12. Two consecutive paragraphs begin with "Regarding the Flow index." Please review the wording.

Pp12. “dilettantish fluid” or “dilatant fluid”?

Pp13. Viscoelastic properties - Temperature sweep. Figure 5 needs more details. First, the color assigned to each module should be clarified in the figure caption. Second, it should be specified which curve corresponds to the heating phase and which to the cooling phase. Without this information, it is not possible to follow the discussion.

Pp13. “progressive exudation” by “progressive lixiviation”.

Pp 14. “where aw is less than 0.85”. Lower?

Pp 15. Conclusions. “The method that allows for the highest concentration of protein in sacha inchi is mechanical defatting with a protein result of 73%, compared to that obtained by the wet method, which resulted in 31%.” This does not belong in the Conclusion section, but rather in the Results and Discussion section.

Pp 15. “However, during the production of sacha flour with the mechanical method, bioactive compounds are lost, which is why lower values of total polyphenols and antioxidant activity are obtained, compared to taro flour.” In this sentence, methods for obtaining Sacha Inchi flour are compared, but it is compared with taro flour. Please review the wording.

Pp 15. “The viscosity profile evaluation determined that the starch contained in taro provides good viscosity characteristics, being this an important ingredient for the development of foods such as creams, pastes, sauces, among others.” This is useful for foods that require a heating process to allow starch gelatinization and hence viscosity development.

Pp15. “the optimum level of inclusion”. In fact, the optimal level will depend on the food that is being formulated.

Pp16. “Although taro’s drying temperature of 60 °C maintains polyphenols and antioxidant activity”. We do not know if they "maintain," as it was not compared against other drying temperatures.

Pp16. “using gluten-free orphan raw materials”. What do authors mean by "orphan"?

Is the work clearly and accurately presented and does it cite the current literature?

Yes
Is the study design appropriate and is the work technically sound?

Yes
Are sufficient details of methods and analysis provided to allow replication by others?

Yes
If applicable, is the statistical analysis and its interpretation appropriate?

Yes
Are all the source data underlying the results available to ensure full reproducibility?

Yes
Are the conclusions drawn adequately supported by the results?

Partly

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

Gluten free bakery, starch properties

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Responses (1)

Author Response

07 Mar 2025

Karen Muñoz, Departamento de Agroindustria, Universidad del Cauca, Popayan, Colombia

Dear Dr. Lorena Sciarini,

We would like to express our sincere gratitude for the valuable contributions you made during the evaluation process of our article. Your suggestions have been highly enriching, and we have taken them into account in order to enhance the quality and clarity of our work. Below we
respond to each of the suggestions made.

Pp1. Abstract. “The ABTS radical method, which reacts with the phenolic compounds of the food matrix, using Trolox as a standard.” Please review the wording (the sentence is incomplete).

Response: Thank you for your observation. We have reviewed and corrected the wording of the sentence to make it clearer and more complete, while also complementing the main idea. The revised version is as follows: “Methods: Proximal tests were conducted to determine basic chemical composition, quantification of free polyphenols, antioxidant activity using the
ABTS* radical method (2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)) with Trolox as a standard, and rheological analyses, including pasting curves, flow profiles, and viscoelastic properties. Microbiological characterization of the flours was also performed.”

Pp 2. Does "Salmonella" correspond to a specific species? Please revise (it could also be Salmonella sp. or Salmonella spp.).

Response: We have reviewed the entire document and replaced 'Salmonella' with the correct designation 'Salmonella spp.' to refer to the corresponding species.

Pp2. It is suggested to change “According to Ref. 1” to “According to Wang et al.¹.” Consider applying this modification throughout the text.

Response: Thank you for your observation. We have taken your suggestion into account and have adjusted the referencing format throughout the text accordingly

Pp 4. Please, replace “mNicroorganisms” with “microorganisms”

Response: Thank you very much for the observation. It has been replaced with the appropriate term “microorganisms”.

Pp4. “this, in order to grind the grain. Obtaining a milky and homogeneous” Please revise the punctuation (I understand the period is not appropriate).

Response: The suggestion was taken into account, and the wording was improved to enhance the clarity of the text. The revised version is as follows: “The sacha inchi flour
obtained by the wet method (SIF-WM) was prepared as follows: initially, wet grinding was carried out using a blender (Oster, BLST 4655, Colombia) with a capacity of 1.25 L. For this process, sacha inchi seeds were mixed with water in a 1:3 ratio to ensure effective grinding. This resulted in a milky and homogeneous suspension. Subsequently, a cloth filter was used to separate the insoluble extract (cake) from the water-soluble extract (milk). The cake was
then dried in an oven at 65 °C for 3 hours. Finally, once dehydrated, it was immediately packaged in a polyethylene bag with a hermetic seal for preservation [18].”

Pp4. Deffating method. Was the extraction cake dried? How was it milled afterward? Was it sieved? Please complete the methodology.

Response: La requested information was completed and the text's wording was improved. The modified version is as follows: “The defatted sacha inchi flour (SIF-DM) was obtained as follows: the sacha inchi kernels were placed in the hopper of an automatic touchscreen oil extraction press (CGLDENWALL, model K28, Shanghai, China), as shown in Figure 1. The process was carried out at a temperature of 124 °C. During the extraction, a screw conveyor pushed the material into the main pressing cylinder. As it moved forward, the design of the screw, with decreasing pitch and spiral depth, reduced the available volume in the chamber,
subjecting the material to high pressure and friction against the closed bottom wall of the cylinder. This facilitated efficient oil extraction, with the oil flowing through the cylinder's orifices, while the residual material, known as cake, was expelled through the discharge nozzle. Subsequently, the cake was ground using an electric mill (Quaker City Mill, model 4-E, Philadelphia) and sieved through a system with a No. 30 mesh to achieve uniform particle size. Finally, the resulting flour was packaged in polyethylene bags with airtight seals for preservation.”

Pp5. For the proximate composition, please specify which AOAC method was used in each case.

Response: The provided suggestion was taken into account, and in response, the AOAC method used for each compositional analysis was included in each case.

Protein: “For the determination of protein content, the protocol established by AOAC 968.06 was followed.”

Fiber: “The fiber content was analyzed according to the AOAC 962.09 protocol.”

Ash: “ The ash content was determined following the protocol established by AOAC 942.05.”

Pp6. Ref. 20 is incomplete in the References section. Please review all citations.

Response: All references have been reviewed and completed where necessary.

Pp 6. Polyphenols. “Finally, the supernatant mixture was made up to 20 mL using distilled water and the absorbance was measured in the spectrophotometer”. Which colorimetric technique was used? Folin-Ciocalteu? Please clarify.

Response: The raised concern was taken into account, and the wording was complemented and improved to ensure greater clarity, resulting in the following enhanced versión: “For the second extraction, 8 mL of 70% acetone with 1% formic acid (70 mL acetone + 30 mL water + 1 mL formic acid) was added to the pellet from the first extraction. The pellet was shaken in the solution for 25 minutes in the shaker at 200 RPM at room temperature. Subsequently, the
samples were centrifuged at 3500 RPM for 5 minutes at room temperature, and the second supernatant was combined with the first supernatant and shaken. The resulting mixture was made up to 20 mL using distilled water. Next, the measurement was carried out using the Folin-Ciocalteu method. 20 µL of the extract was taken and combined with 900 µL of Folin reagent, vortexed, and then 600 µL of NaHCO3 was added, shaking again to ensure proper dissolution. The reaction was allowed to rest, and finally, the absorbance was measured at 765 nm in the spectrophotometer (Thermo Scientific, Genesys 10S UV VIS).”

Pp6. Ref. 20 is incomplete in the References section. Please review all citations.

Response: All references have been reviewed and completed where necessary.

Pp 6. Polyphenols. “Finally, the supernatant mixture was made up to 20 mL using distilled water and the absorbance was measured in the spectrophotometer”. Which colorimetric technique was used? Folin-Ciocalteu? Please clarify.

Response: The raised concern was taken into account, and the wording was complemented and improved to ensure greater clarity, resulting in the following enhanced versión: “For the second extraction, 8 mL of 70% acetone with 1% formic acid (70 mL acetone + 30 mL water + 1 mL formic acid) was added to the pellet from the first extraction. The pellet was shaken in the solution for 25 minutes in the shaker at 200 RPM at room temperature. Subsequently, the
samples were centrifuged at 3500 RPM for 5 minutes at room temperature, and the second supernatant was combined with the first supernatant and shaken. The resulting mixture was made up to 20 mL using distilled water. Next, the measurement was carried out using the Folin-Ciocalteu method. 20 µL of the extract was taken and combined with 900 µL of Folin reagent, vortexed, and then 600 µL of NaHCO3 was added, shaking again to ensure proper dissolution. The reaction was allowed to rest, and finally, the absorbance was measured at 765 nm in the spectrophotometer (Thermo Scientific, Genesys 10S UV VIS).”

Pp6. ABTS. “In a test tube 4mL of ABTS.* (2,20-Azino-bis(3-ethylbenzthiazoline-6-sulfonic acid), solution was added and to start the reaction 135 μL standard solution and vortexed for 5 seconds.” Please revise the wording.

Response: The suggestion made was taken into account, and the wording was improved, resulting in the following: “In a test tube, 4 mL of ABTS.* solution (2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) was added, followed by 135 μL of the standard solution, and it was vortexed for 5 seconds. For the reagent blank, a mixture of 4 mL of 4.5 acetate buffer and 135 μL of ethanol was used. The zero point was prepared with 4 mL of ABTS solution and 135 μL of ethanol. The tube was then capped and allowed to rest for exactly 30 minutes before measuring the absorbance in the spectrophotometer (Thermo Scientific, Genesys 10S UV VIS) at a wavelength of 729 nm.”

Pp6. Flow profile. Which geometry was used?

Response: The question raised was answered, and the wording was improved. The modified version is as follows: “The flow properties were determined following the methodology outlined by Polo and Roa [21], with some modifications. An AR1500 rheometer (TA Instruments, New Castle, USA) was used for this study, and measurements were taken using cylindrical geometry. The average viscosity was determined at 25 °C for 12 minutes, with the shear rate
increasing in 4 steps as follows:…)

Pp7. Fungi and yeast. “30°” with “30 °C”.

Response: Thank you for the observation, the corresponding correction has already been made.

Pp8. Results and discussion. Table 1. The wet method of obtaining Sacha inchi flours results in the solubilization of hydrophilic compounds, such as proteins. This causes the concentration of other components to increase (particularly carbohydrates, according to the authors' findings). However, it is striking that the lipid concentration is so similar, given that the mechanical method specifically extracts the oil. What is the reason for this similarity?

Response: The similarity in lipid concentration between the flours obtained through the wet and mechanical methods can be explained by the specific characteristics of each process. In the wet method, sacha inchi seeds are mixed with water, and while there is no initial loss of solids, some hydrophilic compounds, such as proteins and carbohydrates, migrate into the
solvent during the soaking and homogenization stages. During filtration, the cake retains the lipids, while soluble compounds are removed with the liquid byproduct. Additionally, the cake obtained through the wet method does not undergo the compression applied by the mechanical press, meaning the necessary pressure to fully extract the oil is not achieved. As a result, a greater amount of lipids is retained in the final cake from the wet method, which explains the similarity in lipid concentration values between the two methods.

Pp8. “As for carbohydrate content, SIF-DM showed a lower percentage of carbohydrates compared to other extraction methods, due to the fact that in the oil extraction process, not only fat is removed but also some solids such as proteins, minerals, and certain carbohydrates (see Figure 1B). These components may become trapped in the oil or be eliminated with the byproduct generated during extraction. Additionally, it is likely that some soluble carbohydrates dissolve or are removed along with the oil during mechanical extraction”. It is more likely that carbohydrates are solubilized in the water used in the wet method rather than in the oil, although some carryover may occur. It is worth exploring the fact that the wet method actually has a higher carbohydrate content because, proportionally, it has fewer proteins, leading to
a concentration of carbohydrates (which are calculated by difference).

Response: Thank you very much for the observations. We took the suggestions into account, resulting in the following version: ” Regarding carbohydrate content, SIF-DM showed a lower percentage compared to other extraction methods. This can be explained by the fact that during the mechanical oil extraction process, not only fat is removed but also some solids, such as proteins, minerals, and certain carbohydrates, which could become trapped in the oil or be eliminated along with the byproducts generated during extraction (see Figure 1B). On the other hand, it is likely that carbohydrates are solubilized in the water used in the wet method, contributing to their higher content in SIF-WM. Additionally, the higher carbohydrate content observed in SIF-WM could be due to a proportional effect: the reduction in proteins during the wet process increases the relative proportion of carbohydrates, which are calculated by difference. Moreover, the wet method may promote the retention of insoluble
carbohydrates within the solid matrix, minimizing the loss of hydrophilic compounds and explaining the higher carbohydrate content in SIF-WM.”

Pp9. Was the proximate composition of the flour blends measured or calculated?

Response: The wording was adjusted based on the observation made. Below, the revised and corrected version is presented: “The study design was based on the type of flour and its inclusion levels in the mixture, considering protein content and carbohydrate percentage as the most relevant factors in each treatment. The proximal composition of the blends was calculated using the individual results obtained from each flour. The analyses showed that the 25% SIF-DM:75% TF blend contained 22.69% protein and 64.83% carbohydrates; the 50% SIF-DM:50% TF blend showed 39.34% protein and 44.23% carbohydrates; and the 75% SIF-DM:25% TF blend reached 55.98% protein and 23.64% carbohydrates. On the other hand, the blends with SIF-WM presented the following values: 25% SIF-WM:75% TF with 12.42%
protein and 73.47% carbohydrates; 50% SIF-WM:50% TF with 18.80% protein and 61.52% carbohydrates; and finally, the 75% SIF-WM:25% TF blend with 25.17% protein and 49.56% carbohydrates.”

Pp9. Effect of processing on FPP and the antioxidant activities. Please do not repeat in the text the values already presented in Tables/Figures. Verify this throughout the manuscript. Also, check, as "taro flour presents the highest polyphenol content of 7.50+/-1.73" does not match the value presented in Table 2.

Response: We have taken the suggestions into account and corrected the corresponding values. The modified version is as follows: “Table 2 highlights statistically significant differences in the total polyphenol content among the flours analyzed. Taro flour exhibited the highest polyphenol content, aligning with findings reported by Eleazu et al. [42] and surpassing other food sources such as plantain flour, as noted by [43]. Meanwhile, defatted sacha inchi flour demonstrated a higher polyphenol content compared to the flour obtained through the wet method. This result is likely due to the mechanical extraction process, which involves pressure and temperature, facilitating the release of phenolic compounds, as previously observed [44].”

Pp 10. “This difference can be explained because taro is a prime source of antioxidant” What is meant by "prime source"?

Response: We aimed to replace the term 'primary source' with a more context-appropriate expression, resulting in the following revision: “Regarding the antioxidant activity measured using the ABTS method, taro flour demonstrated significantly higher activity compared to SIF-DM and SIF-WM. This difference can be attributed to taro being a prominent source of antioxidant compounds and containing various biologically active phytoconstituents, such as flavonoids, sterols, and glycosides [45].”

Pp10. “… bioactive compounds, which in sacha inchi are present particularly in polyunsaturated essential fatty acids…” ABTS is used to evaluate the radical scavenging capacity of hydrophilic compounds. In fact, the extraction was performed with polar solvents. I find the explanation contradictory.

Response: You are absolutely right; the explanation was contradictory. Therefore, we have
revised the analysis and discussion of results, resulting in the following versión. “In this study, a drying temperature of 60 °C was used for the taro flour (TF), which allows for achieving the maximum antioxidant activity in taro flours [46]. The SIF-DM showed the lowest antioxidant activity, which can be attributed to the loss of polar hydrophilic compounds, such as polyphenols, during the mechanical extraction. These compounds are the main contributors to
the radical scavenging capacity measured using the ABTS

Pp11. “to the post hoc Tukey analysis five different groups identified with letters from “a” to “e” are classified. In group “a”, the samples with the highest viscosity (25% SIF-DM:75% TF and 100% TF) with values of 3.08 Pa.s and 2.99 Pa.s respectively, and in group “e”, the samples with the lowest viscosity value (75% SIF-WM:25% TF and 100% SIF-WM) with values of 0.35 Pa.s and 0.64 Pa.s respectively.” This type of expression
used throughout the manuscript makes the reading tedious and is repeated with what
is presented in graphs/tables. Please simplify by focusing on the statistical differences presented between samples.

Response: We took the suggestion into account throughout the manuscript and simplified some expressions to make the reading more fluid and improve its quality

Pp12. In this page, the results are presented in a disorganized manner. It is suggested to first address Figure 2, and then Figure 3.

Response: Thank you for your suggestion. We have organized the results as you recommended and rewritten them to improve clarity.

Pp12. “According to the above, as shown in Figure 3, the samples that presented a higher viscosity drop are the samples with higher TF content, this is because the crystalline structure (amylopectin) that makes up the granules is altered, when amylose and amylopectin are hydrated individually in excess water, both macromolecules tend to reorganize in a similar structure to the one they had initially.” The authors discuss the minimum viscosity (or trough), but I believe there is some confusion here, as this
parameter is not associated with retrogradation but with gelatinization. The granules absorb water up to a maximum, and with shear at high temperature, they break, decreasing viscosity. Retrogradation occurs when the temperature
decreases.

Response: The suggestion was taken into account, the information was organized, and the wording was improved, resulting in the following structured idea: “In graph 3.A, significant differences in viscosity peaks among the different blends are shown. Samples with higher TF content exhibit the highest viscosity peaks due to the presence of starch. This behavior is attributed to the greater amount of water absorbed and the subsequent swelling of starch granules during heating.

Figure 3.B shows the behavior of the samples before the onset of retrogradation, that is, the viscosity drop occurring at the end of the constant temperature phase before cooling begins. Samples with higher starch content and lower protein content exhibited a more pronounced viscosity drop. This behavior occurs because once starch reaches its maximum expansion capacity, it releases retained water and solubilizes amylose and amylopectin fractions. Subsequently, during cooling, these macromolecules hydrate individually in excess water and reorganize into a structure similar to the original one [51].”

Pp12. “As shown in Figure 3, mixtures with higher inclusion of starch present in TF and less protein contributed by sacha inchi result in more free water for starch molecules to swell and solubilize presenting higher viscosity; on the contrary, at high proportions of protein its interaction with starch granules could stabilize the amylose leaching
channel of starch granules, which resulted in lower swelling power and solubility.” It would be useful to consider, in the mixtures, the effect of dilution of the starch present in TF. On the other hand, proteins also develop viscosity, which could explain why TF+SIF-DM has higher viscosity than TF+SIF-WM.

Response: The observation was taken into account, and the text was adjusted as follows: “…Conversely, in blends with a higher protein content, proteins interact with starch granules, stabilizing the amylose leaching channel and reducing both swelling and solubility of starch. This behavior is influenced by the dilution of starch in taro flour (TF), as a lower starch concentration in the blend limits its ability to absorb water and form viscous structures [52]. Additionally, proteins play a crucial role in viscosity development, which explains why TF+SIF-DM blends exhibit higher viscosity compared to TF+SIF-WM blends. In TF+SIF-DM blends, the preservation of functional proteins promotes more effective interaction with water and starch molecules, resulting in higher viscosity. In contrast, in TF+SIF-WM blends, the loss of proteins during the wet extraction process reduces their contribution to viscosity, thereby lowering the rheological performance of the blend.”

Pp12. “until the amorphous part is solubilized, decreasing viscosity”. In general, when referring to starch in water, the term "dispersion" is used, not "solubilization."

Response: Thank you for your observation. We have replaced the term 'solubilization' with 'dispersion' to refer to starch in water.

Pp12. Flow profile analysis. Please, unify: “flowability index” or “flow index”.

Response: Within the paragraph, we unified the term "flow index", thank you for the observation.

Pp12. Two consecutive paragraphs begin with "Regarding the Flow index." Please review the wording.

Response: Thank you for the suggestion; we took your recommendation into account and improved the writing throughout the text.

Pp12. “dilettantish fluid” or “dilatant fluid”?

Response: Thank you for the observation; we changed the term "diletent fluid" to the correct term "dilatant fluid".

Pp13. Viscoelastic properties - Temperature sweep. Figure 5 needs more details. First, the color assigned to each module should be clarified in the figure caption. Second, it should be specified which curve corresponds to the heating phase and which to the cooling phase. Without this information, it is not possible to follow the discussion.

Response: You are absolutely right, Figure 5 needs more details. Therefore, we have specified the color assigned to each modulus and clearly indicated which curve corresponds to both the heating phase and the cooling phase.

Pp13. “progressive exudation” by “progressive lixiviation”.

Response: You are absolutely right, the change was made using the correct term: progressive lixiviation. “There is a progressive lixiviation of amylose in the swollen granules, where gel-like linkages are formed. Researchers have reported the effect of amylose to increase the firmness of gels during cooling as one of the initial causes of gel firmness. Thus, during the cooling stage, the moduli stay constant with the predominance of the elastic modulus [59].”

Pp 14. “where aw is less than 0.85”. Lower?

Response: Thank you for the observation. The requested change has been made by using the term "below" instead of "lower," which improves the accuracy of the wording. The modified version is as follows:

Table 2 shows that in foods such as sacha inchi flour and taro flour, where water activity is below 0.85, Salmonella spp. and other pathogenic bacteria, such as Staphylococcus, can survive in a viable but non-culturable state for extended periods due to their increased resistance to thermal processes. However, although microorganism growth is not sustained at low water activities, foodborne bacteria and fungi can easily contaminate the flour and survive for extended periods with a reduced reproduction rate [24].

Pp 15. Conclusions. “The method that allows for the highest concentration of protein in sacha inchi is mechanical defatting with a protein result of 73%, compared to that obtained by the wet method, which resulted in 31%.” This does not belong in the Conclusion section, but rather in the Results and Discussion section.

Response: You are right, this statement “The method that allows for the highest concentration of protein in sacha inchi is mechanical defatting with a protein result of 73%, compared to that obtained by the wet method, which resulted in 31%” does not belong in the conclusions section, so we have taken your suggestion into account and have properly relocated it.

Pp 15. “However, during the production of sacha flour with the mechanical method, bioactive compounds are lost, which is why lower values of total polyphenols and antioxidant activity are obtained, compared to taro flour.” In this sentence, methods for obtaining Sacha Inchi flour are compared, but it is compared with taro flour. Please review the wording.

Response: Thank you for the observation. We have improved the wording based on the provided recommendation. “Mechanical defatting stood out as the most efficient method
for obtaining high-protein flours from sacha inchi. However, this process was observed to reduce the preservation of bioactive compounds, decreasing antioxidant activity and total polyphenol content compared to the wet milling method.”

Pp 15. “The viscosity profile evaluation determined that the starch contained in taro provides good viscosity characteristics, being this an important ingredient for the development of foods such as creams, pastes, sauces, among others.” This is useful for foods that require a heating process to allow starch gelatinization and hence viscosity development.

Response: We took the suggestion into account and have modified it as follows: “The
viscosity profile evaluation determined that the starch present in taro provides good viscosity characteristics, making it an important ingredient for the development of foods such as creams, pastes, sauces, among others. These properties are particularly useful in foods that require a heating process to allow starch gelatinization and, therefore, viscosity development. However, mixtures with an excess of taro have a higher rate of retrogradation, which represents a disadvantage for food stability during storage.”

Pp15. “the optimum level of inclusion”. In fact, the optimal level will depend on the food that is being formulated.

Response: You are absolutely right; the term used was not appropriate. Therefore, we have attached an improved version of the conclusion below: “Through the viscoelasticity analysis
(temperature sweep), it was determined that an inclusion level of defatted sacha inchi flour between 25% and 50% results in a stable and consistent gel, which could be utilized in various food applications.”

Pp16. “Although taro’s drying temperature of 60 °C maintains polyphenols and antioxidant activity”. We do not know if they "maintain," as it was not compared against other drying temperatures.

Response: You are absolutely right; the term used was not appropriate. Therefore, we have attached an improved version of the conclusion below: “The drying temperature of taro at 60 °C preserves polyphenols and antioxidant activity at significant levels; however, it does not allow the inactivation of bacteria such as Salmonella spp., which has shown resistance to thermal treatment in recent years.”

Pp16. “using gluten-free orphan raw materials”. What do authors mean by "orphan"?

Response: We have replaced the term 'orphan' and improved the wording of this conclusion: “ The development of foods using gluten-free and underutilized raw materials, such as taro and sacha inchi, which easily adapt to regions facing food security challenges, represents a promising alternative for offering the market highly nutritious and gluten-free products.”

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Competing Interests

No competing interests were disclosed.

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Reviewer Report

9 Views

23 Nov 2024 | for Version 3

Elisa Julianti, Department of Food Science Faculty of Agriculture, Universitas Sumatera Utara, Medan, North Sumatra, Indonesia

9 Views Cite this report Responses(0)

Approved

Thank you for the opportunity to review the revised article and the author responses. I have no further comments to make at this time.

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

Flour and Starch from Root and Tuber Crops as Alternatives to Wheat Flour

I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard.

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Reviewer Report

13 Views

17 Sep 2024 | for Version 2

Umi Purwandari, University of Trunojoyo Madura, Madura, Indonesia

13 Views Cite this report Responses(1)

Approved With Reservations

Is the work clearly and accurately presented and does it cite the current literature?

Partly
Is the study design appropriate and is the work technically sound?

Yes
Are sufficient details of methods and analysis provided to allow replication by others?

Partly
If applicable, is the statistical analysis and its interpretation appropriate?

Yes
Are all the source data underlying the results available to ensure full reproducibility?

Yes
Are the conclusions drawn adequately supported by the results?

Yes

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

Food science, food microbiology

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Author Response

22 Nov 2024

Karen Muñoz, Departamento de Agroindustria, Universidad del Cauca, Popayan, Colombia

Dear Dr Purwandari, Thank you very much for your thoughtful and constructive feedback on our manuscript. We appreciate the time and effort you dedicated to reviewing our work, as well as your insightful suggestions, which have greatly helped us improve our paper. Below, we address each of your comments in detail:

1.The paper was written carefully enough and data were analyzed thoroughly using
recent related publication. However, microbiological analysis did not seem relevant
to the work, and therefore it would be better deleted.
Regarding the microbiological analysis, it is important to note that this was conducted to
evaluate the initial state of the obtained flours concerning their microbiological
composition. Since these flours are intended for food consumption, it is essential to know
the level of microbial growth and to ensure that they fall within the permissible ranges
established by current regulations governing such products.

2. Some parts were not correctly written, such as in page 10 where it was refer to
Table2 for oil concentration. While Table 2 was about antioxidant activity. Also,
statistical notation for Table 2, ABTS did not look right.

With the purpose of addressing the question raised, the information was supplemented
using the correct term ;granulometry.; Additionally, the article mentions the following
textually “Comparing SIF-DM flour with SIF-WM, the defatting method produces a
flour with higher viscosity due to the interaction of water with proteins and starch,
considering that SIF-DM has 72.62% protein. This result can be attributed to the fact
that the lower viscosity of sacha inchi flour obtained by the wet method is due to the
sample;s particle size (sieving with a standard mesh size of 425 to 600 microns), as it
was not possible to reduce particle size due to its high fat content, and the resulting
granules from the liquefaction and subsequent drying processes were used.
Therefore, the endogenous lipids present in the taro flour form a type of protective
layer around the aggregate of carbohydrates and proteins, preventing water
absorption and the subsequent swelling of starch”.

3 Some parts were not correctly written, such as in page 10 where it was refer to
Table2 for oil concentration. While Table 2 was about antioxidant activity. Also,
statistical notation for Table 2, ABTS did not look right.
Regarding the suggested corrections, these have been properly adjusted, where Table 1 was
referenced and the scientific notation was modified.

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Competing Interests

No competing interests were disclosed.

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Reviewer Report

20 Views

10 Sep 2024 | for Version 2

Elisa Julianti, Department of Food Science Faculty of Agriculture, Universitas Sumatera Utara, Medan, North Sumatra, Indonesia

20 Views Cite this report Responses(1)

Approved With Reservations

Is the work clearly and accurately presented and does it cite the current literature?

Yes
Is the study design appropriate and is the work technically sound?

Partly
Are sufficient details of methods and analysis provided to allow replication by others?

Partly
If applicable, is the statistical analysis and its interpretation appropriate?

No
Are all the source data underlying the results available to ensure full reproducibility?

Yes
Are the conclusions drawn adequately supported by the results?

Yes

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

Flour and Starch from Root and Tuber Crops as Alternatives to Wheat Flour

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Responses (1)

Author Response

19 Nov 2024

Karen Muñoz, Departamento de Agroindustria, Universidad del Cauca, Popayan, Colombia

Dear Dr Elisa,Thank you very much for your thoughtful and constructive feedback on our manuscript. We appreciate the time and effort you dedicated to reviewing our work, as well as your insightful suggestions, which have greatly helped us improve our paper. Below, we address each of your comments in detail:

1. In conclusion of abstract, Based on the characteristics obtained, what type of food is suitable for the application of sacha inchi and taro flour? For example, would it be bread, pasta, biscuits, or other types of products?

In response to Question 1, the conclusion of the document has been adjusted to mention that, based on the obtained characteristics, “the analyzed flours represent a raw material with great potential for the development of products such as protein snacks due to the protein contribution of sacha inchi, good expansion due to taro starch, and being naturally gluten-free for individuals with celiac disease”.

2. In the introduction, it should be justified what are the drawbacks of using sacha inchi and taro as single food ingredients? Why do you want to combine sacha inchi and taro flour as food ingredients?
In response to Question 2, the introduction has been supplemented with information about some drawbacks or limitations of both sacha inchi and taro. “Sacha inchi can be unstable due to its high concentration of unsaturated fatty acids, making it sensitive to oxidation. Additionally, sacha inchi has limiting amino acids, such as lysine and tryptophan, which may cause low digestibility. Likewise, this seed contains antinutritional factors that limit its use as a raw material in the development of new products. On the other hand, taro contains antinutrients such as oxalates, which can restrict its fresh consumption due to the irritation they cause in the throat and mouth. However, the extrusion process significantly reduces these issues while simultaneously enhancing nutritional properties, such as increased protein digestibility. Based on this, combining sacha inchi and taro is an excellent option to complement their nutritional properties, increasing nutritional value and improving the stability and texture of the final product”.

3. In the methods : The method does not describe the analytical procedure for sacha inchi almonds. Was any treatment applied to the seeds before analysis? Is there a reason for using the ratio of SIF and TF?In relation to Question 3, in the section on methods used to obtain sacha inchi flour, additional information is provided about the process followed prior to the proximal characterization of the sacha inchi kernel. It is explained verbatim that “the sacha inchi seeds were received as white kernels, meaning without shell or husk. For the compositional analysis of the kernel, they were manually ground with a mortar to reduce their size, thus facilitating the relevant tests”. Furthermore, the reason for using the SIF and TF ratio lies in their high nutritional value (carbohydrates and protein) and the fact that both raw materials have had limited scientific and industrial advancement, presenting a significant opportunity to leverage their potential.

4. 4. In the discussion :
- It should be explained what causes the difference in protein content obtained through
the wet method and the mechanical defatting method.

To address this question, additional information has been included to explain the cause of
differences in protein content obtained through the wet milling method compared to the mechanical defatting method. The document states verbatim: “It is important to note that the protein content obtained through the wet milling method is lower compared to the mechanical defatting method. This is because, during the wet milling process, a milky suspension is generated, which is then filtered to separate the insoluble extract (cake) from the water-soluble extract (slurry). During filtration, part of the protein may remain in the soluble extract, which explains the lower protein concentration in the cake. In this study, the proximate analysis was performed only on the insoluble extract”.

- Sacha inchi flour obtained through the wet method is flour derived from the dried,
water-insoluble components. The water-soluble carbohydrate components are certainly not
included in SIF. However, why is the carbohydrate content in sacha inchi almonds lower
than in SIF-DM? Additionally, why is the carbohydrate content in SIF-DM lower, even
though only the fat is removed during its production? Please explain.

Regarding the question, why is the carbohydrate content in sacha inchi kernels lower than
in SIF-DM? Table 1 shows that the kernel contains 6.67% carbohydrates, while SIF-DM
showed 3.04% carbohydrates. In relation to the question of why the carbohydrate content in
SIF-DM is lower, despite the fact that only fat is removed during its production: as shown
in the following image, it is important to mention that during the oil extraction process, not
only fat is removed but also some solids such as proteins, minerals, and some
carbohydrates. These components may become trapped in the oil or be removed with the
byproduct generated during extraction. Additionally, it is likely that some soluble
carbohydrates dissolve or are removed along with the oil during mechanical extraction.

- In SIF-DM, there is a release of phenolic compounds due to involving pressure and
temperature, but why is the FPP content in SIF-DM higher than in SIF-WM? Does the
released FPP not get lost during the processing?

Regarding the question of why the free total phenol content (FPP) in SIF-DM is higher than
in SIF-WM, it is important to consider the differences in the extraction processes. During
the wet milling process (SIF-WM), phenolic compounds, which are water-soluble, can
migrate into the liquid solvent during the maceration stage, resulting in a significant loss of
these bioactive compounds in the solvent and thus decreasing the concentration of phenols
in the final flour. In contrast, the mechanical defatting process (SIF-DM), a more physical
method that does not involve liquid solvents, results in much lower phenolic compound
loss. Without a significant aqueous phase, the phenols remain largely within the solid
matrix of the flour.

- It would be more meaningful if the researcher could show the starch content of each
sample with different SIF and TF ratios.

To complement this information in the article, the following text has been added detailing the
carbohydrate percentage and protein content in each mixture with different inclusion levels. “The study design was based on the type of flour and its inclusion levels in the mixture, taking protein content and carbohydrate percentage as the most relevant factors in each treatment. The following results were obtained: the 25% SIF-DM:75% TF mixture had 22.69% protein and 64.83%
carbohydrates; the 50% SIF-DM:50% TF mixture showed 39.34% protein and 44.23%
carbohydrates; and the 75% SIF-DM:25% TF mixture yielded 55.98% protein and 23.64%
carbohydrates. Meanwhile, the mixtures with SIF-WM presented the following values: 25% SIF-
WM:75% TF with 12.42% protein and 73.47% carbohydrates; 50% SIF-WM:50% TF with 18.80% protein and 61.52% carbohydrates; and finally, the 75% SIF-WM:25% TF mixture with 25.17% protein and 49.56% carbohydrates”.

- Based on Table 3, the aw for all three types of flour is <0.85, which should prevent the
growth of microorganisms. How do you explain the presence of Salmonella and mold in
your flour products?

As mentioned in Table 3, the analyzed flours exhibit low water activity, with values below 0.85.
This level of water activity is not suitable for the growth of most microorganisms, as the available water is insufficient to sustain basic metabolic processes. However, while low water activity inhibits the active proliferation of microorganisms, it does not guarantee the total absence of pathogens. Spores of certain microorganisms and bacteria, such as Salmonella, can survive under adverse conditions, remaining viable for extended periods. Additionally, many pathogens possess persistence mechanisms that allow them to survive in hostile environments. In this case, Salmonella can form protective structures called biofilms, which enable microorganisms to adhere to surfaces
and resist disinfection treatments. Moreover, these biofilms not only act as physical barriers but also increase the tolerance of microorganisms to physical processes such as drying, protecting bacterial cells from environmental stress.
5. In conclusion : What is the recommended ratio of SIF to TF ?
To address this question, the conclusions state that through the viscoelasticity analysis (temperature sweep), it was determined that the optimal inclusion level of defatted sacha inchi is between 25% and 50%, which allows for the formation of a stable and consistent gel. Complementing this conclusion, the 25% SIF-DM:75% TF mixture has 22.69% protein and 64.83% carbohydrates, while the 50% SIF-DM:50% TF mixture shows 39.34% protein and 44.23% carbohydrates. These values are relevant for the development of products such as extruded snacks, where the protein contribution from sacha inchi is crucial, and the expansion capability provided by taro starch allows both ingredients to complement each other effectively.

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Competing Interests

No competing interests were disclosed.

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Reviewer Report

39 Views

04 Sep 2023 | for Version 1

Diego Salazar, Facultad de Ciencia e Ingeniería en Alimentos, Universidad Tecnica de Ambato, Ambato, Tungurahua, Ecuador; Universidad Complutense de Madrid, Madrid, Community of Madrid, Spain

39 Views Cite this report Responses(2)

Not Approved

Is the work clearly and accurately presented and does it cite the current literature?

No
Is the study design appropriate and is the work technically sound?

Partly
Are sufficient details of methods and analysis provided to allow replication by others?

Partly
If applicable, is the statistical analysis and its interpretation appropriate?

No
Are all the source data underlying the results available to ensure full reproducibility?

No
Are the conclusions drawn adequately supported by the results?

Partly

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

Food Science and Technology. Research in new sources of foods, andean crops, legumes, tubers.

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Reviewer Report

43 Views

04 Sep 2023 | for Version 1

Oluwaseun P Bamidele, University of Venda, Thohoyandou, Limpopo, South Africa

43 Views Cite this report Responses(1)

Not Approved

Is the work clearly and accurately presented and does it cite the current literature?

No
Is the study design appropriate and is the work technically sound?

No
Are sufficient details of methods and analysis provided to allow replication by others?

Yes
If applicable, is the statistical analysis and its interpretation appropriate?

No
Are all the source data underlying the results available to ensure full reproducibility?

Yes
Are the conclusions drawn adequately supported by the results?

No

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

Food Chemistry, Food processing and preservation, Encapsulation of bioactive compounds, Functional foods, Gamma irradiation.

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Author Response

09 Oct 2023

Karen Muñoz, Departamento de Agroindustria, Universidad del Cauca, Popayan, Colombia

The corrections suggested in the pdf of the manuscript were adjusted.

A characterization of the two raw materials taro and sacha inchi is made.

This research was done in order to pave the way for new projects on the development of food based on vegetable matrices.

Mixtures with inclusion levels of 25, 50 and 75% of sacha inchi flour and taro 75, 50 and 25% were worked in order to evaluate the bioactive, rheological and proximal properties.

We kindly request to specify the paragraphs or sentences that should be adjusted to improve the analysis.

In addition, improvements were made in the English language.

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Competing Interests

No competing interests were disclosed.

Alongside their report, reviewers assign a status to the article:

Approved - the paper is scientifically sound in its current form and only minor, if any, improvements are suggested

Approved with reservations - A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit.

Not approved - fundamental flaws in the paper seriously undermine the findings and conclusions

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Characterization of sacha inchi (Plukenetia volubilis) and taro (Colocasia esculenta) flours with potential application in the preparation of both gluten-free and high protein foods.

Abstract

Keywords

Revised Amendments from Version 1

Introduction

Methods

Obtaining taro flour

Obtaining Sacha inchi flour

Figure 1. Sacha inchi defatting process.

Proximate analysis

Determination of free polyphenols, FPP fraction

Determination of antioxidant activity by the ABTS method

Rheological analysis

(1)

Microbiological characterization

Statistical analysis

Results and discussion

Proximate analysis

Table 1. Proximate composition of flours.

Effect of processing on FPP and the antioxidant activities

Table 2. Bioactive properties of papachina and sachainchi flour.

Rheological analysis

Figure 2. Effect of the different blends on the curves of the pasting curves.

Figure 3. Pasting properties of flour blends.

Figure 4. Flow profile of flour blends.

Figure 5. Temperature sweeping, modules G′ and G″ of the flour blends.

Microbiological analysis

Table 3. Microbiological analysis.

Figure 6. Colonies seeding taro flour sample on XLD agar after 24 h.

Conclusions

Data availability

Underlying data

Extended data

References

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