Physicochemical characteristics of <i>Pangasius</i> sp. skin-gelatin-based-edible film enriched with silver nanoparticles

Rahmi Nurdiani; Asep A. Prihanto; Muhamad Firdaus; Faridatul Nur Aini; Fajrin Adin Nabilah; Rosnita A Talib; Nurul Huda

doi:10.12688/f1000research.129024.2

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Research Article

Revised

Physicochemical characteristics of Pangasius sp. skin-gelatin-based-edible film enriched with silver nanoparticles

[version 2; peer review: 1 approved, 1 approved with reservations, 1 not approved]

Rahmi Nurdiani ^1,2, Asep A. Prihanto^1,2, Muhamad Firdaus^1,2, [...] Faridatul Nur Aini¹, Fajrin Adin Nabilah¹, Rosnita A Talib³, Nurul Huda⁴

Rahmi Nurdiani ^1,2, Asep A. Prihanto^1,2, [...] Muhamad Firdaus^1,2, Faridatul Nur Aini¹, Fajrin Adin Nabilah¹, Rosnita A Talib³, Nurul Huda⁴

PUBLISHED 25 Jul 2024

Author details Author details

¹ Fish Product Technology, Universitas Brawijaya, Malang, East java, 65145, Indonesia
² Bioseafood Research Unit, Universitas Brawijaya, Malang, East Java, 65145, Indonesia
³ Department of Process and Food Engineering, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia
⁴ Faculty of Sustainable Agriculture, Universiti Malaysia Sabah, Sandakan, 90509, Sabah, Malaysia

Rahmi Nurdiani
Roles: Conceptualization, Funding Acquisition, Supervision, Writing – Original Draft Preparation, Writing – Review & Editing

Asep A. Prihanto
Roles: Funding Acquisition, Methodology, Supervision, Writing – Review & Editing

Muhamad Firdaus
Roles: Conceptualization, Formal Analysis, Methodology, Supervision

Faridatul Nur Aini
Roles: Data Curation, Investigation, Methodology

Fajrin Adin Nabilah
Roles: Data Curation, Investigation, Methodology

Rosnita A Talib
Roles: Methodology, Writing – Review & Editing

Nurul Huda
Roles: Conceptualization, Methodology, Writing – Review & Editing

OPEN PEER REVIEW

REVIEWER STATUS

This article is included in the Nanoscience & Nanotechnology gateway.

Abstract

Background: Edible films intended for food packaging have been produced from hydrocolloids, lipids, resins, and composites, including gelatin. Gelatin is known to have a good filming ability and has been suggested as an alternative to non-biodegradable plastics. Naturally active compounds incorporated into film packaging may not only protect the food product from oxidation and microbial contamination, but they may also alter the physicochemical properties of the film. Silver nanoparticles have been used in food packaging as active agents due to their antibacterial and antifungal properties. The addition may affect the characteristics of the packaging. Therefore, this study aims to determine the effect of the addition of silver nanoparticles on the physicochemical characteristics of edible film from Pangasius sp. skin gelatin.

Methods: Mangrove extract of Bruguiera gymnorrhiza was used to synthesize silver into nanoparticle size. In this study, silver nanoparticles (AgNPs) with different concentrations (0, 2, 4 and 8%) were added into gelatin-based edible film. The edible films produced were observed for their physicochemical characteristics, including thickness, tensile strength, elongation, water vapour transmission, moisture content, pH, and colour.

Results: AgNPs affected the colour of the fish-gelatin-based edible film, as an increased concentration of AgNPs resulted in a darker film. Nevertheless, the addition of AgNPs showed no significant effect on the thickness (145–216 µm), tensile strength (14.58–19.72 MPa), elongation (21.86–54.19%), water vapour transmission (30.91–42.55 g/m²/day), moisture content (9.57–11.16%) or pH (5.92–6.01) of the fish-gelatin-based edible film.

Conclusions: The addition of AgNPs has no significant effect on gelatin-based edible film physicochemical properties except colour. Therefore, the incorporated edible film has the potential to be developed further.

Keywords

Bruguiera gymnorrhiza, edible film, gelatin, physicochemical characteristics, silver nanoparticles

Corresponding author: Rahmi Nurdiani

Competing interests: No competing interests were disclosed.

Grant information: The study was fully funded through a World Class Research Grant, No: 612.26/UN10.C10/TU/2022
The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Copyright: © 2024 Nurdiani R 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: Nurdiani R, Prihanto AA, Firdaus M et al. Physicochemical characteristics of Pangasius sp. skin-gelatin-based-edible film enriched with silver nanoparticles [version 2; peer review: 1 approved, 1 approved with reservations, 1 not approved]. F1000Research 2024, 12:160 (https://doi.org/10.12688/f1000research.129024.2) First published: 10 Feb 2023, 12:160 (https://doi.org/10.12688/f1000research.129024.1) Latest published: 25 Jul 2024, 12:160 (https://doi.org/10.12688/f1000research.129024.2)

Revised Amendments from Version 1

The new version of the manuscript provides more information, primarily based on the reviewers' comments. In the introduction section, there is a thorough explanation regarding the mangrove Bruguiera gymnorhiza used in the study. Based on the previous research, B. gymnorhiza leaf extract has exhibited significant inhibitory effects against various pathogenic bacteria. It added importance to the use of its extract as an active agent for gelatin-based edible film. The leaves of Bruguiera gymnorrhiza (Taxonomy ID: 39984) were collected from the Sendang Biru coastal area, Malang, East Java, Indonesia.
The results and discussion section described that adding silver nanoparticles, regardless of the concentration, did not significantly affect any of the physicochemical parameters evaluated. This might be because the concentration of silver nanoparticles added was very small. Gutiérrez et al.²² investigated the impact of adding calcium and silver nanoparticles to gelatin-based films and obtained a similar result. They found that the films' structural integrity and mechanical properties were preserved. The dispersion of AgNPs within the gelatin matrix was uniform, preventing significant changes in the film's tensile strength and elasticity. The barrier properties, such as water vapour permeability, remained stable.
Further data analysis showed that 4% AgNPs was recommended to achieve the best qualities in edible film, meeting established standards. The results suggest that AgNPs can be effective active agents for developing active packaging without negatively affecting the film's physicochemical properties. However, further research is necessary to explore the biological functions of edible films with added AgNPs.

See the authors' detailed response to the review by Amiza Mat Amin
See the authors' detailed response to the review by Mohamed Bilal Goudjil
See the authors' detailed response to the review by Abhilash Sasidharan

Introduction

Due to their excellent structural properties and performances, many petrochemical-based plastics are widely used. However, since these materials are not deemed environmentally friendly, increased public awareness has led to the continued development of biodegradable packaging technology.¹ Edible film, one of the biodegradable packaging materials, is a thin layer attached to a food product and can be consumed.² Edible films can be made from various materials such as polysaccharides (e.g.: starch, pectins, chitin and chitosan), lipids (e.g.: bees wax, oils), and protein (e.g.: whey, soy, pea protein, collagen, gelatin).

Gelatin is a protein macromolecule obtained from the hydrolysis of natural collagen found in skin, bone and animal connective tissue.³ Usually, gelatin comes from cows or pigs, but using raw material sources for gelatin from these animals may cause some problems related to health and religious regulations. Alternatively, fish skin is considered a safer option as a raw material for gelatin production.⁴ Pangasius catfish skin is a by-product resulting from the processing of fish fillets. The processing of Pangasius catfish fillets produces up to 55% of by-products, including fish skin.⁵ The skin can potentially be used as a raw material for gelatin due to its abundant availability. Fish gelatin is now commonly used in the production of edible films due to its low melting temperature, low oxygen permeability, and good film-forming ability.⁶

Nowadays, the concept of active packaging has attracted the food packaging industry and researchers. Active packaging may enable the interaction between the packaging, the food product, and the surrounding environment to improve the sensory properties and safety of food products. In addition, active agents with antimicrobial and antioxidant properties can help in extending the food’s shelf life and prevent food oxidation.⁷ The edible film made using fish skin gelatin usually has low biological properties, so enriching it with active compounds is necessary to produce active packaging. Several studies have shown that edible films made from fish gelatin can be manufactured by adding several active ingredients.⁸ One group of active ingredients that have the potential to be added are silver nanoparticles. Silver nanoparticles (AgNPs) are materials with at least one dimension with a size between 1 and 100 nm.⁹ They can be made by synthesizing silver metal into nano-sized particles. This synthesis can be carried out with a biosynthesis biological agent.⁹ In previous studies, the synthesis of silver nanoparticles from mangrove leaves such as Excoecaria agallocha L.¹⁰ and Rhizophora mucronata¹¹^,¹² has been achieved. In this study, the biosynthesis process was done using the extract of mangrove leaves, Bruguiera gymnorrhiza. Bruguiera gymnorhiza, a member of the Rhizophoraceae family, is characterized by its glabrous and relatively smooth trunk with reddish-brown bark. This species is typically found along the seaward margins of mangrove swamps. B. gymnorhiza leaf extract has demonstrated potential as an active compound for incorporation into fish gelatin films, exhibiting inhibitory effects against various pathogenic bacteria such as Pseudomonas aeruginosa (5.77-23.13 mm), Aeromonas hydrophila (14.17-23.47 mm), and Staphylococcus aureus (1.70-4.43 mm).¹³ Additionally, the leaf extract has shown significant radical scavenging activity, with an IC₅₀ value of 12.93 μg/mL.¹⁴ The content of bioactive compounds in plant extracts can be used as a bio-reductant that can convert silver metal into AgNPs.⁹

Nurdiani et al.,¹⁵ stated that AgNPs can improve the characteristics and functionalities of edible films, such as increasing product resistance, improving barrier properties as packaging and antimicrobial compounds. Therefore, this research was conducted to determine the physicochemical characteristics of edible film from Pangasius catfish skin gelatin with the addition of AgNPs of Bruguiera gymnorrhiza.

Methods

Preparation of gelatin

Pangasius sp. skin (frozen) was provided by PT. RUM Seafood, Sidoarjo Indonesia, and delivered to the Faculty of Fisheries and Marine Science, Universitas Brawijaya. Once the skins were thawed, they were cut into small pieces (0.5 × 0.5 cm²) using clean scissors. The skins were stored in a freezer (−20°C) until use. Gelatin manufacturing started by soaking the Pangasius sp. skin in NaOH 0.1 M solution with a ratio of 1:5 (w/v) for two hours at room temperature. Afterwards, the skin was washed with running water until the pH was normal (pH = 7). Next, the skin was then soaked in an acetic acid solution with a concentration of 0.6 M at a ratio of 1:5 (w/v) for two hours (at room temperature). The swollen skin was then extracted with Aqua Dest, with a ratio of 1:3 (w/v) in a water bath (at 55–60°C) for four hours. Afterwards, the filtrate was collected and poured onto a baking tray for drying using a dehydrator for 8 to 12 hours at 55–60°C. The gelatin sheets obtained were then milled using a grinder to make gelatin powder.⁸

Preparation of mangrove extract

The leaves of Bruguiera gymnorrhiza (Taxonomy ID: 39984) were collected from the Sendang Biru coastal area, Malang, East Java, Indonesia. The leaves were dried using a dehydrator for 8–12 hours at a temperature of 40–45°C. Dried leaves were then reduced in size using a grinder. Two grams of mangrove powder was then dissolved into 100 mL of deionized water and heated to boiling for three minutes. The mangrove solution was allowed to cool to room temperature, filtered using a Buchner funnel, and poured into a dark glass bottle.¹²

Preparation of silver nanoparticles (AgNPs)

One mL of mangrove leaf extract was mixed with 9 mL 20 mM AgNO₃. The solutions were incubated for 0, 5, 10, and 15 minutes. The colour change was observed from the original whitish brown to brown.⁹

Production of edible films

Edible film was made by dissolving 4 grams gelatin into 100 mL distilled water. The gelatin solution was then heated at 50°C for 30 minutes. Next, the glycerol was added at 0.5% (v/v) and heated at 45°C for 15 minutes. AgNPs with the best incubation time was added at several concentrations per 100 mL (0 mL, 2 mL, 4 mL, and 8 mL). The concentration of silver nanoparticles (AgNPs) added to the edible film ranged from the minimum amount needed to show antibacterial activity to the maximum amount considered safe.¹⁶^,¹⁷ Once ready, the edible film solution was evenly poured into an 18 cm × 18 cm non-stick baking pan before being dried in an oven at a temperature of 55°C for 18–20 hours. The edible film was then removed from the oven, left at room temperature for 10 minutes, and peeled off slowly.¹⁵

Physicochemical characteristics analyses

Thickness

The thickness of the edible film sample was measured using a micrometre screw with an accuracy of 0.01 mm. Thickness measurements were carried out at five different points, and the average results represented the sample.¹⁸

Moisture content

An empty crucible cup was oven dried at 105°C for 30 minutes. As it cooled down, 1 gram of the sample was then put into the cup and weighed (W1). Next, the sample was put into the oven and dried at a temperature of 105°C for three hours. After that, the cup was weighed again (W2).¹⁹

Moisture content = \frac{W 1 - W 2}{Sample weight} \times 100 %

Water vapour transmission rate (WVTR)

The WVTR of each film was measured according to Ref. 20. The edible film was cut into a circle with a diameter of 3 cm. Ten grams of silica gel were poured into a cup. The sample was glued on top of the cup to cover it. The cup was weighed and recorded as W1. The sample was allowed to stand for 24 hours at room temperature and was then weighed again and recorded as W2. The WVTR was then calculated using the formula:

WVTR = \frac{Weight difference}{Sample surface area} \times Time

Tensile strength and elongation

Edible film was cut to a size of 5 × 1 cm². The thickness and initial length of the sample were measured. Next, each end of the edible film was clamped on a tensile machine. The sample was then pulled at 10 mm/min speed until it broke.¹⁸ Finally, the tensile strength value was calculated by dividing the maximum stress (F max) by the area of the edible film using the formula:

Tensile strength = \frac{F max}{A}

The measurement of the elongation of the edible film was carried out using a tensile strength device. Elongation is expressed as a percentage and is measured using the formula:

Elongation (%) = \frac{Stretch when breaking up (cm)}{Initial length (cm)} \times 100 %

The pH test was carried out using a pH meter. The measurement was conducted on the edible film solution prior to the drying process.²¹

Colour

The colour of the edible films was measured using a colorimeter. Measurements were made by placing an edible film sample on the colour reader sensor. The reading button was set to L* (lightness), a* (redness) and b* (yellowness), and the target button was pressed. The results then appeared on the screen.

Statistical analysis

SPSS commercial software (IBM SPSS Statistics version 25.0, Chicago, IL, USA) was used for statistical analysis. The data were expressed as mean ± standard deviation (mean ± SD). A one-way analysis of variance (ANOVA) and Tukey’s test with a p < 0.05 significance level were applied.

Results and discussion

The physicochemical properties of edible films enriched with various concentrations of silver nanoparticles were evaluated. In this study, silver nanoparticles were synthesized using green synthesis with mangrove leaf extract. The reason for the formation of nanoparticles using green synthesis for active agents incorporated in gelatin edible films is biocompatibility. Green-synthesized nanoparticles are often biocompatible and non-toxic, making them suitable for use in edible films that come into direct contact with food products.²² The green synthesis of nanoparticles using plant extracts involves a mechanism that includes the reduction of metal ions, complex formation, stabilization, and shape control. This process utilizes the natural compounds present in plant extracts to facilitate the synthesis of nanoparticles with unique properties.²³

The addition of silver nanoparticles, regardless of the concentration, did not significantly affect any of the physicochemical parameters evaluated (Table 1). This might be because the concentration of silver nanoparticles added was very small. Similar with the study conducted by Gutiérrez et al.²⁴ which investigated the impact of adding calcium and silver nanoparticles to gelatin-based films. They found that the films' structural integrity and mechanical properties were preserved. The dispersion of AgNPs within the gelatin matrix was uniform, preventing significant changes in the film's tensile strength and elasticity. Additionally, the barrier properties, such as water vapor permeability, remained stable.

Thickness of enriched edible film

Thickness is one of the characteristics of edible coatings that greatly affects the biological properties and shelf life of food-coating products.²⁵ This study showed that the addition of AgNPs into the gelatin solution had no significant effect (p > 0.05) on the thickness of edible film. The average thickness of the edible film was between 145 and 216 μm (Table 1). According to the Japanese Industrial Standard (JIS),²⁶ the maximum thickness of edible film should be 250 μm. The thickness of the edible film is affected by the density, viscosity and surface tension of edible film materials as well as the drying time and layering technique.²⁷

Table 1. Physicochemical characteristics Pangasius sp. skin-gelatin-based edible film incorporated with AgNPs.

No	Conc. of AgNPs	Thickness (μm)	Tensile strength (MPa)	Elongation (%)	Water vapour transmission (g/m²/day)	Moisture content (%)	pH
1	0%	145.50 ± 52.57^a	19.72 ± 11.52^a	21.86 ± 21.72^a	37.09 ± 25.65^a	9.57 ± 1.13^a	6.01 ± 0.16^a
2	2%	149.00 ± 57.51^a	18.36 ± 5.14^a	52.47 ± 41.28^a	30.91 ± 14.46^a	11.16 ± 1.97^a	5.96 ± 0.11^a
3	4%	163.00 ± 33.80^a	15.67 ± 6.58^a	54.19 ± 59.02^a	42.55 ± 18.96^a	10.40 ± 0.72^a	5.95 ± 0.14^a
4	8%	216.00 ± 21.78^a	14.58 ± 5.88^a	36.54 ± 38.35^a	35.32 ± 29.79^a	9.75 ± 1.81^a	5.92 ± 0.15^a

Similarly, Arfat et al.²¹ reported an increasing of edible film thickness due to the addition of silver–copper nanoparticles (Ag–CuNPs). The resulting range was 61–98 μm. The increase in thickness of the edible film was related to the increase of the solid content of the edible film. Furthermore, the addition of Ag–CuNPs to the gelatin affected the structure of the protein, causing it to become regular and the film network to be more robust.

Moisture content of enriched edible film

The moisture content test was crucial to determine the total number of water molecules in the edible film tissue. The moisture content in the edible film affects the stability of the product, so it is expected that the moisture content in the edible film produced is low. This study showed that adding AgNPs into the gelatin solution had no significant effect (p > 0.05) on the moisture content of each edible film. The moisture content of the edible film had an average value of between 9.57 and 11.16% (Table 1). Our result was lower than the study reported by Kanmani & Rhim.²⁸

Adding AgNPs into an edible film will usually increase its moisture content because of reduced interactions between gelatin chains and increased availability of free hydroxyl groups to absorb water. The moisture content of edible films can also be affected by temperature and drying methods. The temperature will affect the time needed in the drying process, causing an increase in the intensity of the interaction between water molecules and the hydrophilic polymer chain.²⁹

WVTR

Statistically, the WVTR test indicated that adding AgNPs into the gelatin solution had no significant effect (p > 0.05) on the value of the WVTR of the edible film. The average value of the WVTR of the edible film produced was 30.91–42.55 g/m²/day (Table 1). Our results did not meet the Japanese Industrial Standard (JIS),²⁶ a maximum WVTR of 7 g/m²/day. The high value of the WVTR of the edible films is due to the fact that the materials used to make the edibles are made of protein-derived materials that have polar polymers and a large number of hydrogen bonds, thus making the edible films capable of absorbing water at high humidity. The factors that can affect the WVTR values are the presence of chemicals and the structure of the constituent materials, the concentration of plasticizers, and environmental conditions such as humidity and temperature.³⁰

Tensile strength and elongation

Table 1 shows that adding AgNPs into the gelatin-based edible film solution had no significant effect (p > 0.05) on the tensile strength of the edible film. The average tensile strength of the edible film was 14.58–19.72 MPa (Table 1). The results are in accordance with the Japanese Industrial Standard (JIS),²⁶ as the minimum tensile strength of edible film should be 0.3 MPa. The value of the tensile strength of the edible films decreased with increasing concentrations of AgNPs. This could be because AgNPs are very weak in forming crosslinks with fish skin gelatin during the production of the edible films. In comparison, the elongation test results showed that adding AgNPs into the gelatin solution had no significant effect (p > 0.05) on the elongation of each edible film. The resulting edible film elongation value ranged from 54.19 to 21.86% (Table 1). The results, however, were still much lower than the standard minimum elongation value of 70.³¹ Elongation is inversely proportional to tensile strength.²³ According to Skurtys et al.,³² elongation is expressed as a percentage change in the film’s original length before breaking and gives a measure of film elongation. Elongation values range from 1 to 80%. The level of elasticity of the edible coating can be seen from the elongation value; the higher the elongation value, the more elastic the edible coating.

The addition of AgNPs into the film resulted in a decrease in the tensile strength value. This might be due to the reduction in protein chain reactions due to incorporation with AgNPs.²⁵ The tensile strength of a film can be influenced by the constituent materials that have structural cohesion properties and the constituent materials of edible films, namely hydrocolloid protein (gelatin), contain amino acids capable of forming disulphide bonds.³³ In addition, the properties of the constituent materials to create a more robust gel network will help the formation of polysaccharide molecular structures more closely, forming a more coherent film structure and reducing the absorption of water molecules.³⁴ Bonilla & Sobral³⁵ stated that the elongation value of gelatin and chitosan edible films with ethanolic extracts from plants increased with the increased volume of extracts. This is due to the interaction between the phenolic compounds present in the extract. Furthermore, gelatin peptides can form covalent crosslinks, forming a more cohesive and flexible matrix. The higher the elongation value of the film, the more elastic it is so that the film can be stretched more. Meanwhile, films with low elongation values will be brittle.

pH

The results showed that the pH of the enriched edible film ranged from 5.92 to 6.01 (Table 1). It was indicated that the higher the concentration of AgNPs Bruguiera gymnorrhiza added, the lower the pH level of fish-gelatin-based edible film. Bruguiera gymnorrhiza mangrove leaf extract was detected to have an acidic pH of around 6.15, thus, the resulting edible film was expected to have a lower pH value than the control (0% AgNPs).

Colour

Colour testing using a colour reader brings up three values: L*, a*, and b*. The L* value indicates the brightness level, a* indicates a red–green colour (a = +60 red, a = -60 green) and b* indicates a yellow–blue colour (b = +60 yellow, b = -60 blue). Edible films produced using gelatin usually have clear or dull/opaque colours.¹⁵ Statistically, the addition of the concentration of AgNPs had a significant effect (p < 0.05) on the L*, a* and b* values (Table 2).

Table 2. Colour value of edible film from Pangasius sp. skin gelatin enriched with AgNPs.

No	Concentration of AgNPs	L*	a*	b*
1	0%	87.363 ± 1.19^c	0.440 ± 0.33^a	1.978 ± 1.89^a
2	2%	82.655 ± 1.39^bc	2.223 ± 0.43^a	7.508 ± 1.58^a
3	4%	74.028 ± 5.41^ab	5.833 ± 1.57^b	13.890 ± 3.19^b
4	8%	70.773 ± 6.36^a	7.398 ± 2.63^b	17.178 ± 3.99^b

The addition of a higher concentration of AgNPs on the fish-gelatin-based edible film resulted in a darker colour (Figure 1), increasing the red and yellowish colour. Similarly, Shankar et al.³⁶ reported that the L* value of edible gelatin films with the addition of metallic nanoparticles (38.20) was lower than the control (93.15).

Figure 1. Appearance of enriched fish-gelatin-based edible film.

The best treatment

The best treatment of edible film made from Pangasius skin gelatin with the addition of silver nanoparticles synthesized using Bruguiera gymnorrhiza extract was determined using the De Garmo method (grouping and weighting parameters, where the weights given are in accordance with the importance/priority of each parameter in affecting the results of the research).³⁷ Based on the calculation, it was suggested that the best treatment for all parameters was the third treatment (the addition of 4% silver nanoparticles) (Table 3).

Table 3. Physicochemical characteristics for the best treatment.

No	Parameter	Edible film	References
1	Thickness (μm)	163.0 ± 33.80	Max 250³¹
2	Tensile strength (MPa)	15.67 ± 6.58	Min 0.3³¹
3	Elongation (%)	54.19 ± 59.02	Min 70³¹
4	WVTR (g/m²/day)	42.55 ± 18.96	Max 5³¹
5	Moisture content (%)	10.40 ± 0.72	Max 13³¹
6	pH	5.95 ± 0.14	5.75–6.60³⁸
7	L* value	74.028 ± 5.41	38.20³⁶
8	a* value	5.83 ± 1.57	15.08³⁶
9	b* value	13.89 ± 3.19	12.24³⁶

Conclusion

The addition of various concentrations of AgNPs (0, 2, 4, and 8%) synthesized using mangrove Bruguiera gymnorrhiza had a significant effect on the colour of Pangasius gelatin-based edible film (L* value 70.773–87.363, a* value 0.440–7.398 and b* value 1.978–17.178). Nevertheless, adding AgNPs did not significantly affect the other physicochemical parameters (thickness, tensile strength, elongation, water vapour transmission, moisture content, and pH). This study recommends using 4% AgNPs to achieve the best qualities in edible film, meeting established standards. The results suggest that AgNPs can be effective active agents for developing active packaging without negatively affecting the film's physicochemical properties. However, further research is needed to explore the biological functions of edible films with added AgNPs.

Ethical approval

This study did not require ethical approval because the authors used fish skins (available as waste or by-products) from the fish processing industry. Mangrove leaves were collected from a local mangrove conservation area in Malang, East Java. The conservation officer carried out the identification and collection of the mangrove leaves. The process was conducted in a way to ensures the sustainability of the local mangrove forest.

Data availability

Underlying data

Figshare: Physicochemical Characteristics of Pangasius sp. Skin Gelatin-based Edible Film Enriched with Silver Nanoparticles. https://doi.org/10.6084/m9.figshare.21640196.v2.³⁸

The project contains the following underlying data:

− DATA F1000 R NURDIANI.xlsx (raw data for colour, thickness, moisture content, water vapour transmission rate, pH, and tensile strength and elongation).

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

Acknowledgements

The authors would like to thank Universitas Brawijaya and the Ministry of Education, Culture, Research and Technology, the Republic of Indonesia, for the research grant.

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Version 2

VERSION 2 PUBLISHED 10 Feb 2023

Author details Author details

Rahmi Nurdiani
Roles: Conceptualization, Funding Acquisition, Supervision, Writing – Original Draft Preparation, Writing – Review & Editing

Asep A. Prihanto
Roles: Funding Acquisition, Methodology, Supervision, Writing – Review & Editing

Muhamad Firdaus
Roles: Conceptualization, Formal Analysis, Methodology, Supervision

Faridatul Nur Aini
Roles: Data Curation, Investigation, Methodology

Fajrin Adin Nabilah
Roles: Data Curation, Investigation, Methodology

Rosnita A Talib
Roles: Methodology, Writing – Review & Editing

Nurul Huda
Roles: Conceptualization, Methodology, Writing – Review & Editing

Competing interests

No competing interests were disclosed.

Grant information

The study was fully funded through a World Class Research Grant, No: 612.26/UN10.C10/TU/2022
The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Article Versions (2)

version 2

Revised

Published: 25 Jul 2024, 12:160

https://doi.org/10.12688/f1000research.129024.2

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Published: 10 Feb 2023, 12:160

https://doi.org/10.12688/f1000research.129024.1

© 2024 Nurdiani R 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.

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Nurdiani R, Prihanto AA, Firdaus M et al. Physicochemical characteristics of Pangasius sp. skin-gelatin-based-edible film enriched with silver nanoparticles [version 2; peer review: 1 approved, 1 approved with reservations, 1 not approved]. F1000Research 2024, 12:160 (https://doi.org/10.12688/f1000research.129024.2)

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Open Peer Review

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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

VERSION 2

PUBLISHED 25 Jul 2024

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Reviewer Report 20 Sep 2024

Abhilash Sasidharan, Norwegian University of Science and Technology (NTNU), Trondheim, Norway; Kerala University of Fisheries and Ocean Studies, Kerala, India

Not Approved

https://doi.org/10.5256/f1000research.168310.r319877

The study is significant considering the topic selected. The replacement of synthetic plastics in food packaging is important in connection to the sustainable developmental goals targeted to be achieved in near future.

This study focusses on the effect of silver nanoparticles on a fish gelatine based edible film. The study concludes that the AgNPs had no significant effect on any of the physicochemical properties.

AgNPs are well known for their antimicrobial properties which is not discussed in this study.

As the study proposes edible application, the toxicity of the film also needs to be discussed in details. Noting that the application percentage is as high as 8%, it needs to be either discussed in detail or demonstrated using toxicity evaluation.

All the values are statistically reported as homogenous with no significant effect. The thickness of the films compared to the control film has increased 32% in 8% AgNP incorporated film. Similar differences in values are evident in many other parameters as well.

Hence the statement “This study showed that the addition of AgNPs into the gelatin solution had no significant effect (p > 0.05) on the thickness of edible film” needs to be reevaluated.

The article needs to be modified regarding the significant level statements and the statistical analysis has to be redone.

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?

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?

No source data required
Are the conclusions drawn adequately supported by the results?

Partly

Competing Interests: No competing interests were disclosed.

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.

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Author Response 19 Nov 2024

Rahmi Nurdiani, Bioseafood Research Unit, Universitas Brawijaya, Malang, 65145, Indonesia

19 Nov 2024

Author Response

Responses to Reviewer 3

Comment 1
AgNPs are well known for their antimicrobial properties which is not discussed in this study.

Response 1
This study primarily focuses on ... Continue reading Responses to Reviewer 3

Comment 1
AgNPs are well known for their antimicrobial properties which is not discussed in this study.

Response 1
This study primarily focuses on examining the physicochemical properties of AgNPs as a foundational investigation. Given its preliminary nature, the research does not cover the antimicrobial properties, despite the known effectiveness of AgNPs in this area. Instead, this study aims to establish baseline data on physicochemical characteristics, paving the way for future studies to delve into antimicrobial effects and applications.

Comment 2
As the study proposes edible application, the toxicity of the film also needs to be discussed in details. Noting that the application percentage is as high as 8%, it needs to be either discussed in detail or demonstrated using toxicity evaluation.

Response 2
As this study is preliminary and focuses primarily on the physicochemical properties of the film, a toxicity test was not included in the scope of this research. The primary objective was to establish baseline data on the film’s properties, which is essential groundwork before moving into applications and safety assessments. Future research is necessary to investigate toxicity in detail, particularly for edible applications, and this study paves the way for such in-depth evaluations.

Comment 3
All the values are statistically reported as homogenous with no significant effect. The thickness of the films compared to the control film has increased 32% in 8% AgNP incorporated film. Similar differences in values are evident in many other parameters as well. Hence the statement “This study showed that the addition of AgNPs into the gelatin solution had no significant effect (p > 0.05) on the thickness of edible film” needs to be reevaluated.

Response 3
Even though the trends in thickness and other parameters appear different—such as the observed 32% increase in thickness in the 8% AgNP-incorporated film compared to the control—the statistical analysis indicated that the differences are not significant (p > 0.05). This suggests that, despite visible variations, the changes are likely due to natural variability and do not reflect a consistent, reliable effect of AgNP incorporation on film thickness.
Responses to Reviewer 3

Comment 1
AgNPs are well known for their antimicrobial properties which is not discussed in this study.

Response 1
This study primarily focuses on examining the physicochemical properties of AgNPs as a foundational investigation. Given its preliminary nature, the research does not cover the antimicrobial properties, despite the known effectiveness of AgNPs in this area. Instead, this study aims to establish baseline data on physicochemical characteristics, paving the way for future studies to delve into antimicrobial effects and applications.

Comment 2
As the study proposes edible application, the toxicity of the film also needs to be discussed in details. Noting that the application percentage is as high as 8%, it needs to be either discussed in detail or demonstrated using toxicity evaluation.

Response 2
As this study is preliminary and focuses primarily on the physicochemical properties of the film, a toxicity test was not included in the scope of this research. The primary objective was to establish baseline data on the film’s properties, which is essential groundwork before moving into applications and safety assessments. Future research is necessary to investigate toxicity in detail, particularly for edible applications, and this study paves the way for such in-depth evaluations.

Comment 3
All the values are statistically reported as homogenous with no significant effect. The thickness of the films compared to the control film has increased 32% in 8% AgNP incorporated film. Similar differences in values are evident in many other parameters as well. Hence the statement “This study showed that the addition of AgNPs into the gelatin solution had no significant effect (p > 0.05) on the thickness of edible film” needs to be reevaluated.

Response 3
Even though the trends in thickness and other parameters appear different—such as the observed 32% increase in thickness in the 8% AgNP-incorporated film compared to the control—the statistical analysis indicated that the differences are not significant (p > 0.05). This suggests that, despite visible variations, the changes are likely due to natural variability and do not reflect a consistent, reliable effect of AgNP incorporation on film thickness.
Competing Interests: I disclose that there is no competing interests Close
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COMMENTS ON THIS REPORT

Author Response 19 Nov 2024

Rahmi Nurdiani, Bioseafood Research Unit, Universitas Brawijaya, Malang, 65145, Indonesia

19 Nov 2024

Author Response

Responses to Reviewer 3

Comment 1
AgNPs are well known for their antimicrobial properties which is not discussed in this study.

Response 1
This study primarily focuses on ... Continue reading Responses to Reviewer 3

Comment 1
AgNPs are well known for their antimicrobial properties which is not discussed in this study.

Response 1
This study primarily focuses on examining the physicochemical properties of AgNPs as a foundational investigation. Given its preliminary nature, the research does not cover the antimicrobial properties, despite the known effectiveness of AgNPs in this area. Instead, this study aims to establish baseline data on physicochemical characteristics, paving the way for future studies to delve into antimicrobial effects and applications.

Comment 2
As the study proposes edible application, the toxicity of the film also needs to be discussed in details. Noting that the application percentage is as high as 8%, it needs to be either discussed in detail or demonstrated using toxicity evaluation.

Response 2
As this study is preliminary and focuses primarily on the physicochemical properties of the film, a toxicity test was not included in the scope of this research. The primary objective was to establish baseline data on the film’s properties, which is essential groundwork before moving into applications and safety assessments. Future research is necessary to investigate toxicity in detail, particularly for edible applications, and this study paves the way for such in-depth evaluations.

Comment 3
All the values are statistically reported as homogenous with no significant effect. The thickness of the films compared to the control film has increased 32% in 8% AgNP incorporated film. Similar differences in values are evident in many other parameters as well. Hence the statement “This study showed that the addition of AgNPs into the gelatin solution had no significant effect (p > 0.05) on the thickness of edible film” needs to be reevaluated.

Response 3
Even though the trends in thickness and other parameters appear different—such as the observed 32% increase in thickness in the 8% AgNP-incorporated film compared to the control—the statistical analysis indicated that the differences are not significant (p > 0.05). This suggests that, despite visible variations, the changes are likely due to natural variability and do not reflect a consistent, reliable effect of AgNP incorporation on film thickness.
Responses to Reviewer 3

Comment 1
AgNPs are well known for their antimicrobial properties which is not discussed in this study.

Response 1
This study primarily focuses on examining the physicochemical properties of AgNPs as a foundational investigation. Given its preliminary nature, the research does not cover the antimicrobial properties, despite the known effectiveness of AgNPs in this area. Instead, this study aims to establish baseline data on physicochemical characteristics, paving the way for future studies to delve into antimicrobial effects and applications.

Comment 2
As the study proposes edible application, the toxicity of the film also needs to be discussed in details. Noting that the application percentage is as high as 8%, it needs to be either discussed in detail or demonstrated using toxicity evaluation.

Response 2
As this study is preliminary and focuses primarily on the physicochemical properties of the film, a toxicity test was not included in the scope of this research. The primary objective was to establish baseline data on the film’s properties, which is essential groundwork before moving into applications and safety assessments. Future research is necessary to investigate toxicity in detail, particularly for edible applications, and this study paves the way for such in-depth evaluations.

Comment 3
All the values are statistically reported as homogenous with no significant effect. The thickness of the films compared to the control film has increased 32% in 8% AgNP incorporated film. Similar differences in values are evident in many other parameters as well. Hence the statement “This study showed that the addition of AgNPs into the gelatin solution had no significant effect (p > 0.05) on the thickness of edible film” needs to be reevaluated.

Response 3
Even though the trends in thickness and other parameters appear different—such as the observed 32% increase in thickness in the 8% AgNP-incorporated film compared to the control—the statistical analysis indicated that the differences are not significant (p > 0.05). This suggests that, despite visible variations, the changes are likely due to natural variability and do not reflect a consistent, reliable effect of AgNP incorporation on film thickness.
Competing Interests: I disclose that there is no competing interests Close
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Reviewer Report 08 Aug 2024

Amiza Mat Amin, Faculty of Fisheries and Food Science, Universiti Malaysia Terengganu, Kuala Nerus, Malaysia

Approved

https://doi.org/10.5256/f1000research.168310.r306972

The authors have ... Continue reading

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Version 1

VERSION 1

PUBLISHED 10 Feb 2023

Views

Reviewer Report 28 May 2024

Amiza Mat Amin, Faculty of Fisheries and Food Science, Universiti Malaysia Terengganu, Kuala Nerus, Malaysia

Approved with Reservations

https://doi.org/10.5256/f1000research.141678.r166792

The manuscript described the effect of incorporating silver nanoparticles in the silver catfish gelatin film. The introduction is sufficient. Please state why the range of 2-8% was used in this study. However, this study did not include antimicrobial activity of ... Continue reading

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?

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?

Yes

Competing Interests: No competing interests were disclosed.

Reviewer Expertise: food science, food protein

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.

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Report a concern

Author Response 25 Jul 2024

Rahmi Nurdiani, Bioseafood Research Unit, Universitas Brawijaya, Malang, 65145, Indonesia

25 Jul 2024

Author Response

Responses to Reviewer 2

Comment 1:
Please state why the range of 2-8% was used in this study. However, this study did not include antimicrobial activity of the gelatine ... Continue reading Responses to Reviewer 2

Comment 1:
Please state why the range of 2-8% was used in this study. However, this study did not include antimicrobial activity of the gelatine films.
Response 1:
The concentration of silver nanoparticles (AgNPs) added to the edible film ranged from the minimum amount needed to show antibacterial activity to the maximum amount considered safe.

Comment 2:
Please comment why there was no significant difference in most results.
Response 2:
The physicochemical properties of edible films enriched with various concentrations of silver nanoparticles were evaluated. The addition of silver nanoparticles, regardless of the concentration, did not significantly affect any of the physicochemical parameters evaluated (Table 1). This might be because the concentration of silver nanoparticles added was very small. Similar to the study conducted by Gutiérrez et al. 24, which investigated the impact of adding calcium and silver nanoparticles to gelatin-based films. They found that the films' structural integrity and mechanical properties were preserved. The dispersion of AgNPs within the gelatin matrix was uniform, preventing significant changes in the film's tensile strength and elasticity. Additionally, the barrier properties, such as water vapour permeability, remained stable.
Responses to Reviewer 2

Comment 1:
Please state why the range of 2-8% was used in this study. However, this study did not include antimicrobial activity of the gelatine films.
Response 1:
The concentration of silver nanoparticles (AgNPs) added to the edible film ranged from the minimum amount needed to show antibacterial activity to the maximum amount considered safe.

Comment 2:
Please comment why there was no significant difference in most results.
Response 2:
The physicochemical properties of edible films enriched with various concentrations of silver nanoparticles were evaluated. The addition of silver nanoparticles, regardless of the concentration, did not significantly affect any of the physicochemical parameters evaluated (Table 1). This might be because the concentration of silver nanoparticles added was very small. Similar to the study conducted by Gutiérrez et al. 24, which investigated the impact of adding calcium and silver nanoparticles to gelatin-based films. They found that the films' structural integrity and mechanical properties were preserved. The dispersion of AgNPs within the gelatin matrix was uniform, preventing significant changes in the film's tensile strength and elasticity. Additionally, the barrier properties, such as water vapour permeability, remained stable.
Competing Interests: No competing interests Close
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COMMENTS ON THIS REPORT

Author Response 25 Jul 2024

Rahmi Nurdiani, Bioseafood Research Unit, Universitas Brawijaya, Malang, 65145, Indonesia

25 Jul 2024

Author Response

Responses to Reviewer 2

Comment 1:
Please state why the range of 2-8% was used in this study. However, this study did not include antimicrobial activity of the gelatine ... Continue reading Responses to Reviewer 2

Comment 1:
Please state why the range of 2-8% was used in this study. However, this study did not include antimicrobial activity of the gelatine films.
Response 1:
The concentration of silver nanoparticles (AgNPs) added to the edible film ranged from the minimum amount needed to show antibacterial activity to the maximum amount considered safe.

Comment 2:
Please comment why there was no significant difference in most results.
Response 2:
The physicochemical properties of edible films enriched with various concentrations of silver nanoparticles were evaluated. The addition of silver nanoparticles, regardless of the concentration, did not significantly affect any of the physicochemical parameters evaluated (Table 1). This might be because the concentration of silver nanoparticles added was very small. Similar to the study conducted by Gutiérrez et al. 24, which investigated the impact of adding calcium and silver nanoparticles to gelatin-based films. They found that the films' structural integrity and mechanical properties were preserved. The dispersion of AgNPs within the gelatin matrix was uniform, preventing significant changes in the film's tensile strength and elasticity. Additionally, the barrier properties, such as water vapour permeability, remained stable.
Responses to Reviewer 2

Comment 1:
Please state why the range of 2-8% was used in this study. However, this study did not include antimicrobial activity of the gelatine films.
Response 1:
The concentration of silver nanoparticles (AgNPs) added to the edible film ranged from the minimum amount needed to show antibacterial activity to the maximum amount considered safe.

Comment 2:
Please comment why there was no significant difference in most results.
Response 2:
The physicochemical properties of edible films enriched with various concentrations of silver nanoparticles were evaluated. The addition of silver nanoparticles, regardless of the concentration, did not significantly affect any of the physicochemical parameters evaluated (Table 1). This might be because the concentration of silver nanoparticles added was very small. Similar to the study conducted by Gutiérrez et al. 24, which investigated the impact of adding calcium and silver nanoparticles to gelatin-based films. They found that the films' structural integrity and mechanical properties were preserved. The dispersion of AgNPs within the gelatin matrix was uniform, preventing significant changes in the film's tensile strength and elasticity. Additionally, the barrier properties, such as water vapour permeability, remained stable.
Competing Interests: No competing interests Close
Report a concern

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Reviewer Report 10 Oct 2023

Mohamed Bilal Goudjil, Applied Sciences Faculty, Process Engineering Laboratory, KASDI Merbah-Ouargla University, Ouargla, Algeria

Approved with Reservations

https://doi.org/10.5256/f1000research.141678.r208387

This is interesting original research that aims to investigate the Physicochemical characteristics of Pangasius sp. skin-gelatin-based-edible film enriched with silver nanoparticles.

Overall, the manuscript is well complied and experimental data is appreciable. However, a few comments below must be addressed by the authors with utmost attention before considering for indexing.

1. In introduction section: Please give some information about the plant used in the present study.

2. Author should provide herbarium number or plant code for the Bruguiera gymnorrhiza used.

3. In the Results and discussion section, a brief talk about the reasons for the formation of nanoparticles using green synthesis, specifying the mechanism and the responsible component, is required.

4. The conclusion is a bit poor, it should be improved to reflect the work done.

5. I believe the language can be more polished by the authors.

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?

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

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

No source data required
Are the conclusions drawn adequately supported by the results?

No

Competing Interests: No competing interests were disclosed.

Reviewer Expertise: Phytochemistry, Essential oil, Chromatography, Renewable Energies, Nanotechnology

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Report a concern

Author Response 25 Jul 2024

Rahmi Nurdiani, Bioseafood Research Unit, Universitas Brawijaya, Malang, 65145, Indonesia

25 Jul 2024

Author Response

Comments from the Reviewers

1^st REVIEWER

Comment 1: In introduction section: Please give some information about the plant used in the present study.
Response 1:
Bruguiera gymnorhiza, a member of ... Continue reading Comments from the Reviewers

1^st REVIEWER

Comment 1: In introduction section: Please give some information about the plant used in the present study.
Response 1:
Bruguiera gymnorhiza, a member of the Rhizophoraceae family, is characterized by its glabrous and relatively smooth trunk with reddish-brown bark. This species is typically found along the seaward margins of mangrove swamps. B. gymnorhiza leaf extract has demonstrated potential as an active compound for incorporation into fish gelatin films, exhibiting inhibitory effects against various pathogenic bacteria such as Pseudomonas aeruginosa (5.77 - 23.13 mm), Aeromonas hydrophila (14.17 - 23.47 mm), and Staphylococcus aureus (1.70 - 4.43 mm) (Effendi et al., 2023). Additionally, the leaf extract has shown significant radical scavenging activity, with an IC₅₀ value of 12.93 µg/mL (Indriaty et al., 2023).

Comment 2: Author should provide herbarium number or plant code for the Bruguiera gymnorrhiza used.
Response 2: The leaves of Bruguiera gymnorrhiza (Taxonomy ID: 39984) were obtained from the Sendang Biru coastal area, Malang, East Java, Indonesia.

Comment 3:
In the Results and discussion section, a brief talk about the reasons for the formation of nanoparticles using green synthesis, specifying the mechanism and the responsible component, is required.
Response 3:
The physicochemical properties of edible films enriched with various concentrations of silver nanoparticles were evaluated. In this study, silver nanoparticles were synthesized using green synthesis with mangrove leaf extract. The reason for the formation of nanoparticles using green synthesis for active agents incorporated in gelatin edible films is biocompatibility. Green-synthesized nanoparticles are often biocompatible and non-toxic, making them suitable for use in edible films that come into direct contact with food products.22 The green synthesis of nanoparticles using plant extracts involves a mechanism that includes the reduction of metal ions, complex formation, stabilization, and shape control. This process utilizes the natural compounds present in plant extracts to facilitate the synthesis of nanoparticles with unique properties.23

Comment 4: The conclusion is a bit poor; it should be improved to reflect the work done.
Response 4:
The addition of various concentrations of AgNPs (0, 2, 4, and 8%) synthesized using mangrove Bruguiera gymnorrhiza had a significant effect on the colour of Pangasius gelatin-based edible film (L* value 70.773–87.363, a* value 0.440–7.398 and b* value 1.978–17.178). Nevertheless, adding AgNPs did not significantly affect the other physicochemical parameters (thickness, tensile strength, elongation, water vapour transmission, moisture content, and pH). This study recommends using 4% AgNPs to achieve the best qualities in edible film, meeting established standards. The results suggest that AgNPs can be effective active agents for developing active packaging without negatively affecting the film's physicochemical properties. However, further research is needed to explore the biological functions of edible films with added AgNPs.

Comment 5:
I believe the language can be more polished by the authors.
Response 5:
Regarding the grammatical and sentence construction errors, we have revised our manuscript.
Comments from the Reviewers

1^st REVIEWER

Comment 1: In introduction section: Please give some information about the plant used in the present study.
Response 1:
Bruguiera gymnorhiza, a member of the Rhizophoraceae family, is characterized by its glabrous and relatively smooth trunk with reddish-brown bark. This species is typically found along the seaward margins of mangrove swamps. B. gymnorhiza leaf extract has demonstrated potential as an active compound for incorporation into fish gelatin films, exhibiting inhibitory effects against various pathogenic bacteria such as Pseudomonas aeruginosa (5.77 - 23.13 mm), Aeromonas hydrophila (14.17 - 23.47 mm), and Staphylococcus aureus (1.70 - 4.43 mm) (Effendi et al., 2023). Additionally, the leaf extract has shown significant radical scavenging activity, with an IC₅₀ value of 12.93 µg/mL (Indriaty et al., 2023).

Comment 2: Author should provide herbarium number or plant code for the Bruguiera gymnorrhiza used.
Response 2: The leaves of Bruguiera gymnorrhiza (Taxonomy ID: 39984) were obtained from the Sendang Biru coastal area, Malang, East Java, Indonesia.

Comment 3:
In the Results and discussion section, a brief talk about the reasons for the formation of nanoparticles using green synthesis, specifying the mechanism and the responsible component, is required.
Response 3:
The physicochemical properties of edible films enriched with various concentrations of silver nanoparticles were evaluated. In this study, silver nanoparticles were synthesized using green synthesis with mangrove leaf extract. The reason for the formation of nanoparticles using green synthesis for active agents incorporated in gelatin edible films is biocompatibility. Green-synthesized nanoparticles are often biocompatible and non-toxic, making them suitable for use in edible films that come into direct contact with food products.22 The green synthesis of nanoparticles using plant extracts involves a mechanism that includes the reduction of metal ions, complex formation, stabilization, and shape control. This process utilizes the natural compounds present in plant extracts to facilitate the synthesis of nanoparticles with unique properties.23

Comment 4: The conclusion is a bit poor; it should be improved to reflect the work done.
Response 4:
The addition of various concentrations of AgNPs (0, 2, 4, and 8%) synthesized using mangrove Bruguiera gymnorrhiza had a significant effect on the colour of Pangasius gelatin-based edible film (L* value 70.773–87.363, a* value 0.440–7.398 and b* value 1.978–17.178). Nevertheless, adding AgNPs did not significantly affect the other physicochemical parameters (thickness, tensile strength, elongation, water vapour transmission, moisture content, and pH). This study recommends using 4% AgNPs to achieve the best qualities in edible film, meeting established standards. The results suggest that AgNPs can be effective active agents for developing active packaging without negatively affecting the film's physicochemical properties. However, further research is needed to explore the biological functions of edible films with added AgNPs.

Comment 5:
I believe the language can be more polished by the authors.
Response 5:
Regarding the grammatical and sentence construction errors, we have revised our manuscript.
Competing Interests: No competing interest Close
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Author Response 25 Jul 2024

Rahmi Nurdiani, Bioseafood Research Unit, Universitas Brawijaya, Malang, 65145, Indonesia

25 Jul 2024

Author Response

Comments from the Reviewers

1^st REVIEWER

Comment 1: In introduction section: Please give some information about the plant used in the present study.
Response 1:
Bruguiera gymnorhiza, a member of ... Continue reading Comments from the Reviewers

1^st REVIEWER

Comment 1: In introduction section: Please give some information about the plant used in the present study.
Response 1:
Bruguiera gymnorhiza, a member of the Rhizophoraceae family, is characterized by its glabrous and relatively smooth trunk with reddish-brown bark. This species is typically found along the seaward margins of mangrove swamps. B. gymnorhiza leaf extract has demonstrated potential as an active compound for incorporation into fish gelatin films, exhibiting inhibitory effects against various pathogenic bacteria such as Pseudomonas aeruginosa (5.77 - 23.13 mm), Aeromonas hydrophila (14.17 - 23.47 mm), and Staphylococcus aureus (1.70 - 4.43 mm) (Effendi et al., 2023). Additionally, the leaf extract has shown significant radical scavenging activity, with an IC₅₀ value of 12.93 µg/mL (Indriaty et al., 2023).

Comment 2: Author should provide herbarium number or plant code for the Bruguiera gymnorrhiza used.
Response 2: The leaves of Bruguiera gymnorrhiza (Taxonomy ID: 39984) were obtained from the Sendang Biru coastal area, Malang, East Java, Indonesia.

Comment 3:
In the Results and discussion section, a brief talk about the reasons for the formation of nanoparticles using green synthesis, specifying the mechanism and the responsible component, is required.
Response 3:
The physicochemical properties of edible films enriched with various concentrations of silver nanoparticles were evaluated. In this study, silver nanoparticles were synthesized using green synthesis with mangrove leaf extract. The reason for the formation of nanoparticles using green synthesis for active agents incorporated in gelatin edible films is biocompatibility. Green-synthesized nanoparticles are often biocompatible and non-toxic, making them suitable for use in edible films that come into direct contact with food products.22 The green synthesis of nanoparticles using plant extracts involves a mechanism that includes the reduction of metal ions, complex formation, stabilization, and shape control. This process utilizes the natural compounds present in plant extracts to facilitate the synthesis of nanoparticles with unique properties.23

Comment 4: The conclusion is a bit poor; it should be improved to reflect the work done.
Response 4:
The addition of various concentrations of AgNPs (0, 2, 4, and 8%) synthesized using mangrove Bruguiera gymnorrhiza had a significant effect on the colour of Pangasius gelatin-based edible film (L* value 70.773–87.363, a* value 0.440–7.398 and b* value 1.978–17.178). Nevertheless, adding AgNPs did not significantly affect the other physicochemical parameters (thickness, tensile strength, elongation, water vapour transmission, moisture content, and pH). This study recommends using 4% AgNPs to achieve the best qualities in edible film, meeting established standards. The results suggest that AgNPs can be effective active agents for developing active packaging without negatively affecting the film's physicochemical properties. However, further research is needed to explore the biological functions of edible films with added AgNPs.

Comment 5:
I believe the language can be more polished by the authors.
Response 5:
Regarding the grammatical and sentence construction errors, we have revised our manuscript.
Comments from the Reviewers

1^st REVIEWER

Comment 1: In introduction section: Please give some information about the plant used in the present study.
Response 1:
Bruguiera gymnorhiza, a member of the Rhizophoraceae family, is characterized by its glabrous and relatively smooth trunk with reddish-brown bark. This species is typically found along the seaward margins of mangrove swamps. B. gymnorhiza leaf extract has demonstrated potential as an active compound for incorporation into fish gelatin films, exhibiting inhibitory effects against various pathogenic bacteria such as Pseudomonas aeruginosa (5.77 - 23.13 mm), Aeromonas hydrophila (14.17 - 23.47 mm), and Staphylococcus aureus (1.70 - 4.43 mm) (Effendi et al., 2023). Additionally, the leaf extract has shown significant radical scavenging activity, with an IC₅₀ value of 12.93 µg/mL (Indriaty et al., 2023).

Comment 2: Author should provide herbarium number or plant code for the Bruguiera gymnorrhiza used.
Response 2: The leaves of Bruguiera gymnorrhiza (Taxonomy ID: 39984) were obtained from the Sendang Biru coastal area, Malang, East Java, Indonesia.

Comment 3:
In the Results and discussion section, a brief talk about the reasons for the formation of nanoparticles using green synthesis, specifying the mechanism and the responsible component, is required.
Response 3:
The physicochemical properties of edible films enriched with various concentrations of silver nanoparticles were evaluated. In this study, silver nanoparticles were synthesized using green synthesis with mangrove leaf extract. The reason for the formation of nanoparticles using green synthesis for active agents incorporated in gelatin edible films is biocompatibility. Green-synthesized nanoparticles are often biocompatible and non-toxic, making them suitable for use in edible films that come into direct contact with food products.22 The green synthesis of nanoparticles using plant extracts involves a mechanism that includes the reduction of metal ions, complex formation, stabilization, and shape control. This process utilizes the natural compounds present in plant extracts to facilitate the synthesis of nanoparticles with unique properties.23

Comment 4: The conclusion is a bit poor; it should be improved to reflect the work done.
Response 4:
The addition of various concentrations of AgNPs (0, 2, 4, and 8%) synthesized using mangrove Bruguiera gymnorrhiza had a significant effect on the colour of Pangasius gelatin-based edible film (L* value 70.773–87.363, a* value 0.440–7.398 and b* value 1.978–17.178). Nevertheless, adding AgNPs did not significantly affect the other physicochemical parameters (thickness, tensile strength, elongation, water vapour transmission, moisture content, and pH). This study recommends using 4% AgNPs to achieve the best qualities in edible film, meeting established standards. The results suggest that AgNPs can be effective active agents for developing active packaging without negatively affecting the film's physicochemical properties. However, further research is needed to explore the biological functions of edible films with added AgNPs.

Comment 5:
I believe the language can be more polished by the authors.
Response 5:
Regarding the grammatical and sentence construction errors, we have revised our manuscript.
Competing Interests: No competing interest Close
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Version 2

VERSION 2 PUBLISHED 10 Feb 2023

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	1	2	3
Version 2 (revision) 25 Jul 24		read	read
Version 1 10 Feb 23	read	read

Mohamed Bilal Goudjil, KASDI Merbah-Ouargla University, Ouargla, Algeria
Amiza Mat Amin, Universiti Malaysia Terengganu, Kuala Nerus, Malaysia
Abhilash Sasidharan, Norwegian University of Science and Technology (NTNU), Trondheim, Norway; Kerala University of Fisheries and Ocean Studies, Kerala, India

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14 Views

20 Sep 2024 | for Version 2

Abhilash Sasidharan, Norwegian University of Science and Technology (NTNU), Trondheim, Norway; Kerala University of Fisheries and Ocean Studies, Kerala, India

14 Views Cite this report Responses(1)

Not Approved

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?

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?

No source data required
Are the conclusions drawn adequately supported by the results?

Partly

Competing Interests

No competing interests were disclosed.

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Author Response

19 Nov 2024

Rahmi Nurdiani, Bioseafood Research Unit, Universitas Brawijaya, Malang, 65145, Indonesia

Responses to Reviewer 3

Comment 1
AgNPs are well known for their antimicrobial properties which is not discussed in this study.

Response 1
This study primarily focuses on examining the physicochemical properties of AgNPs as a foundational investigation. Given its preliminary nature, the research does not cover the antimicrobial properties, despite the known effectiveness of AgNPs in this area. Instead, this study aims to establish baseline data on physicochemical characteristics, paving the way for future studies to delve into antimicrobial effects and applications.

Comment 2
As the study proposes edible application, the toxicity of the film also needs to be discussed in details. Noting that the application percentage is as high as 8%, it needs to be either discussed in detail or demonstrated using toxicity evaluation.

Response 2
As this study is preliminary and focuses primarily on the physicochemical properties of the film, a toxicity test was not included in the scope of this research. The primary objective was to establish baseline data on the film’s properties, which is essential groundwork before moving into applications and safety assessments. Future research is necessary to investigate toxicity in detail, particularly for edible applications, and this study paves the way for such in-depth evaluations.

Comment 3
All the values are statistically reported as homogenous with no significant effect. The thickness of the films compared to the control film has increased 32% in 8% AgNP incorporated film. Similar differences in values are evident in many other parameters as well. Hence the statement “This study showed that the addition of AgNPs into the gelatin solution had no significant effect (p > 0.05) on the thickness of edible film” needs to be reevaluated.

Response 3
Even though the trends in thickness and other parameters appear different—such as the observed 32% increase in thickness in the 8% AgNP-incorporated film compared to the control—the statistical analysis indicated that the differences are not significant (p > 0.05). This suggests that, despite visible variations, the changes are likely due to natural variability and do not reflect a consistent, reliable effect of AgNP incorporation on film thickness.

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I disclose that there is no competing interests

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1 Views

08 Aug 2024 | for Version 2

Amiza Mat Amin, Faculty of Fisheries and Food Science, Universiti Malaysia Terengganu, Kuala Nerus, Malaysia

1 Views Cite this report Responses(0)

Approved

The authors have done the required correction.

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

food science, food protein

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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16 Views

28 May 2024 | for Version 1

Amiza Mat Amin, Faculty of Fisheries and Food Science, Universiti Malaysia Terengganu, Kuala Nerus, Malaysia

16 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?

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?

Yes

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

food science, food protein

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Author Response

25 Jul 2024

Rahmi Nurdiani, Bioseafood Research Unit, Universitas Brawijaya, Malang, 65145, Indonesia

Responses to Reviewer 2

Comment 1:
Please state why the range of 2-8% was used in this study. However, this study did not include antimicrobial activity of the gelatine films.
Response 1:
The concentration of silver nanoparticles (AgNPs) added to the edible film ranged from the minimum amount needed to show antibacterial activity to the maximum amount considered safe.

Comment 2:
Please comment why there was no significant difference in most results.
Response 2:
The physicochemical properties of edible films enriched with various concentrations of silver nanoparticles were evaluated. The addition of silver nanoparticles, regardless of the concentration, did not significantly affect any of the physicochemical parameters evaluated (Table 1). This might be because the concentration of silver nanoparticles added was very small. Similar to the study conducted by Gutiérrez et al. 24, which investigated the impact of adding calcium and silver nanoparticles to gelatin-based films. They found that the films' structural integrity and mechanical properties were preserved. The dispersion of AgNPs within the gelatin matrix was uniform, preventing significant changes in the film's tensile strength and elasticity. Additionally, the barrier properties, such as water vapour permeability, remained stable.

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No competing interests

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33 Views

10 Oct 2023 | for Version 1

Mohamed Bilal Goudjil, Applied Sciences Faculty, Process Engineering Laboratory, KASDI Merbah-Ouargla University, Ouargla, Algeria

33 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?

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

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

No source data required
Are the conclusions drawn adequately supported by the results?

No

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

Phytochemistry, Essential oil, Chromatography, Renewable Energies, Nanotechnology

Respond to this report

Responses (1)

Author Response

25 Jul 2024

Rahmi Nurdiani, Bioseafood Research Unit, Universitas Brawijaya, Malang, 65145, Indonesia

Comments from the Reviewers

1^st REVIEWER

Comment 1: In introduction section: Please give some information about the plant used in the present study.
Response 1:
Bruguiera gymnorhiza, a member of the Rhizophoraceae family, is characterized by its glabrous and relatively smooth trunk with reddish-brown bark. This species is typically found along the seaward margins of mangrove swamps. B. gymnorhiza leaf extract has demonstrated potential as an active compound for incorporation into fish gelatin films, exhibiting inhibitory effects against various pathogenic bacteria such as Pseudomonas aeruginosa (5.77 - 23.13 mm), Aeromonas hydrophila (14.17 - 23.47 mm), and Staphylococcus aureus (1.70 - 4.43 mm) (Effendi et al., 2023). Additionally, the leaf extract has shown significant radical scavenging activity, with an IC₅₀ value of 12.93 µg/mL (Indriaty et al., 2023).

Comment 2: Author should provide herbarium number or plant code for the Bruguiera gymnorrhiza used.
Response 2: The leaves of Bruguiera gymnorrhiza (Taxonomy ID: 39984) were obtained from the Sendang Biru coastal area, Malang, East Java, Indonesia.

Comment 3:
In the Results and discussion section, a brief talk about the reasons for the formation of nanoparticles using green synthesis, specifying the mechanism and the responsible component, is required.
Response 3:
The physicochemical properties of edible films enriched with various concentrations of silver nanoparticles were evaluated. In this study, silver nanoparticles were synthesized using green synthesis with mangrove leaf extract. The reason for the formation of nanoparticles using green synthesis for active agents incorporated in gelatin edible films is biocompatibility. Green-synthesized nanoparticles are often biocompatible and non-toxic, making them suitable for use in edible films that come into direct contact with food products.22 The green synthesis of nanoparticles using plant extracts involves a mechanism that includes the reduction of metal ions, complex formation, stabilization, and shape control. This process utilizes the natural compounds present in plant extracts to facilitate the synthesis of nanoparticles with unique properties.23

Comment 4: The conclusion is a bit poor; it should be improved to reflect the work done.
Response 4:
The addition of various concentrations of AgNPs (0, 2, 4, and 8%) synthesized using mangrove Bruguiera gymnorrhiza had a significant effect on the colour of Pangasius gelatin-based edible film (L* value 70.773–87.363, a* value 0.440–7.398 and b* value 1.978–17.178). Nevertheless, adding AgNPs did not significantly affect the other physicochemical parameters (thickness, tensile strength, elongation, water vapour transmission, moisture content, and pH). This study recommends using 4% AgNPs to achieve the best qualities in edible film, meeting established standards. The results suggest that AgNPs can be effective active agents for developing active packaging without negatively affecting the film's physicochemical properties. However, further research is needed to explore the biological functions of edible films with added AgNPs.

Comment 5:
I believe the language can be more polished by the authors.
Response 5:
Regarding the grammatical and sentence construction errors, we have revised our manuscript.

View more View less

Competing Interests

No competing interest

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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Physicochemical characteristics of Pangasius sp. skin-gelatin-based-edible film enriched with silver nanoparticles

Abstract

Keywords

Revised Amendments from Version 1

Introduction

Methods

Preparation of gelatin

Preparation of mangrove extract

Preparation of silver nanoparticles (AgNPs)

Production of edible films

Physicochemical characteristics analyses

Statistical analysis

Results and discussion

Thickness of enriched edible film

Table 1. Physicochemical characteristics Pangasius sp. skin-gelatin-based edible film incorporated with AgNPs.

Moisture content of enriched edible film

WVTR

Tensile strength and elongation

pH

Colour

Table 2. Colour value of edible film from Pangasius sp. skin gelatin enriched with AgNPs.

Figure 1. Appearance of enriched fish-gelatin-based edible film.

The best treatment

Table 3. Physicochemical characteristics for the best treatment.

Conclusion

Ethical approval

Data availability

Underlying data

Acknowledgements

References

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