Keywords
Zanthoxylum acanthopodium DC, essential oil isolation, essential oil content, proximate analysis, mineral analysis, GS-MS, secondary metabolites, TLC.
-
Views
-
Downloads
How to cite this article
Satria D, Dalimunthe A, Pertiwi D et al. Phytochemicals, proximate composition, minerals and volatile oil analysis of Zanthoxylum acanthopodium DC. fruits [version 1; peer review: 1 not approved]. F1000Research 2023, 12:227 (https://doi.org/10.12688/f1000research.128941.1)
NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article.
Export Citation
Sciwheel
EndNote
Ref. Manager
Bibtex
ProCite
Sente
▬
✚
Research Article
Phytochemicals, proximate composition, minerals and volatile oil analysis of Zanthoxylum acanthopodium DC. fruits
[version 1; peer review: 1 not approved]
PUBLISHED 01 Mar 2023
OPEN PEER REVIEW
REVIEWER STATUS
This article is included in the Plant Science gateway.
Abstract
Corresponding author:
Denny Satria
Competing interests:
No competing interests were disclosed.
Grant information:
This research was funded by the Ministry of Education, Culture, Research and Technology Republic of Indonesia through the “Hibah Penelitian Dasar Unggulan Perguruan Tinggi” research grant 2021-2023 (Contract No: 059/E5/PG.02.00/PT/2022).
The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Copyright:
© 2023 Satria D 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: Satria D, Dalimunthe A, Pertiwi D et al. Phytochemicals, proximate composition, minerals and volatile oil analysis of Zanthoxylum acanthopodium DC. fruits [version 1; peer review: 1 not approved]. F1000Research 2023, 12:227 (https://doi.org/10.12688/f1000research.128941.1)
First published: 01 Mar 2023, 12:227 (https://doi.org/10.12688/f1000research.128941.1)
Latest published: 01 Mar 2023, 12:227 (https://doi.org/10.12688/f1000research.128941.1)
The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Introduction
Due to their potent antioxidant qualities, lack of side effects, and economic viability, plants have been studied for their medicinal properties in the modern scientific community. In terms of effectiveness and absence of side effects, alternative medicine systems can always be trusted.1 According to World Health Organization (WHO) statistics, over 50,000 plant species are used in traditional medicines around the world, and about 21,000 plants are employed in alternative medicine. The entire plant or different parts, such as the leaves, stem, bark, root, flower, tuber, and seed, among others, are used to make the drugs.2 Over 30% of all plant species have been used for medical purposes at some point in their history. When compared to ethnomedicine's use in practice, conventional medication practice in the treatment of diseases and infections has embraced a more scientific and broad measurement, particularly in developing nations.3
Phytochemicals are just one of the several secondary metabolites that give medicinal plants their therapeutic potential. Recent research on phenolic compounds has highlighted several of their potential medical benefits.4 Andaliman's phenolic compounds can help people with diabetes heal burns by making more of a protein called VEGF (vascular endothelial growth factor)5 and are able to inhibit the development of free radicals, so many tests are carried out as an antioxidant.6 The phenolic mixtures' abilities to scavenge free radicals and lessen lipid peroxidation are linked to their anticarcinogenic, antimutagenic, and cardioprotective effects.7
Zanthoxylum acanthopodium DC is also used to treat dysentery. Andaliman has been utilized by Indians to treat leprosy and skin conditions like abscesses and paralysis.8 In North Sumatera, notably in North Tapanuli, andaliman has been utilized as a spice.7,8 Numerous substances, including phenol hydroquinones, flavonoids, steroids/triterpenoids, tannins, glycosides, volatile oils, alkaloids, coumarins, lignans, amides, and terpenes, are found in plants belonging to the Zanthoxylum genus.9,10 It has been demonstrated that the andaliman fruit ethylacetate extract (EAF) has cytotoxic effects on the MCF-7 and T47D cell lines. It was discovered that doxorubicin and EAF worked in harmony.11 By a cardioprotective impact and action on T47D resistant cells, EAF was demonstrated to have anticancer activity in mice induced with benzo(a) pyrene.11 Given the aforementioned background, the researchers are interested in performing proximate analyses, secondary metabolite components, and essential oil concentration found in Zanthoxylum acanthopodium DC.
Methods
Plant material
The material used in this study was the fresh fruit of Zanthoxylum acanthopodium DC. Green-black in color, round in shape, and weighing 5 grams on average, this fruit was collected in Onan Rugu Village, Samosir Regency, North Sumatra Province, Indonesia.
Preparation of simplicia
Andaliman fruit is washed with running water and then dried at room temperature. After drying, it was ground using a blender. The sample was blended until smooth, until it looked like powder, to get Zanthoxylum acanthopodium DC powder.9
Qualitative phytochemical screening
The identification of secondary metabolites was carried out using the thin-layer chromatography (TLC) method. A total of 100 mg of extract was dissolved in 1 ml ethanol and then smeared on the stationary phase. The stationary phase was a plate coated with silica gel 60 F254 (Merck, Germany) measuring 10x5 cm. The scale was the inserted into a chamber that has been saturated with mobile phase vapor.12,13 Phytochemical screening tests can be seen in Table 1.
Table 1. Qualitative phytochemicals analysis with TLC.
Isolation of volatile oils
The hydrodistillation method was used to isolate essential oils. The Zanthoxylum acanthopodium DC powder was placed in a long-necked flat-bottom flask with distilled water and distilled for 4-5 hours. The essential oil was collected in a separatory funnel, and then the essential oil was separated from the water. 1ml of the anhydrous sodium sulfate was added to the essential oil, shaken, let stand for 1 day, and stored in a dark bottle.14
Analysis of essential oil components
The determination of essential oil components was carried out using a Gas Chromatograph-Mass Spectrometer (GC-MS; MS). An Rtx-5 MS capillary column (Restek) with a column length of 30 meters, a column diameter of 0.25 mm, an injector temperature of 270 C, and a flow rate of 1.16 ml/minute were used in the analysis. The column temperature was programmed (temperature programming) with an initial temperature of 60 C for 5 minutes, then increased slowly at an increase rate of 5.0 C/minute until the final temperature was 280 C, which was maintained for 30 minutes with the ionizing electron impact (EI) type.
The way to identify essential oil components is to compare the mass spectra of the essential oil components obtained with the mass spectra in the data library that have the highest similarity index.10
Proximate analysis
The AOAC method was used to identify to quantify the amounts of ash, water, carbs, total fat, protein, and crude fiber.15 used toy
Water content
The gravimetric method was used to calculate how much water was in the sample. The sample was weighed until the weight stayed the same, which meant that all the water in the sample had evaporated. The procedure for analyzing the water content is that the cup to be used is dried beforehand at a temperature of 100–105°C for 30 minutes. Then it was dried in an oven at 100–150 C for 6 hours and cooled again in a desiccator for 30 minutes.16
Analysis of ash
The ashing method was used to determine the ash content. The principle of this analysis is to burn the material or incinerate it at a high temperature (about 350 °C) and weighing the substance that remains after the ashing.17
Protein content
Protein content was determined using 0.1 g of powder placed in a 100 ml Kjedhal flask. 2 ml of H2SO4 98% was added and 0.9 g selenium as a catalyst, and allowed to digest for 60 minutes, after which a sample was taken. Distillation was then carried out. The distillate obtained was put in an Erlenmeyer flask containing 15 ml of acid solution, borate 4%, and a mixed indicator (2-3 drops of methyl red and methyl blue). The mixed distillate is titrated with a standard solution in the form of 0.02 N HCl, until a light purple color is obtained.18
Lipid content
A total of 5 g of sample was put in a lead filter, then covered with fat-free cotton wool. Lead containing samples were mounted on a soxhlet extraction unit. The weight is filled with sufficient petroleum benzene (30 ml), then installed on the extraction tool. After the extraction is complete (5 hours), the flask is heated in the oven. It is then put in desicator until all of benzene has evaporated.19
Carbohydrate content
An analysis of the carbohydrate content was carried out using a different method in the calculated proximate analysis. Using the formula = 100% -%(moisture content + ash content + fat content + protein content), so that the carbohydrate content of the material is obtained.20
Mineral analysis
NaCl and KCl were used as standards for the mineral analysis, which was done using a flame photometer (Model 405 Corning, UK), after which the sample's mineral content was checked. For all other metals in the sample, an atomic absorption spectrophotometer (Pekin-Elmar Model 403, Norwalk, CT, USA) was used. The determination of each mineral was done twice. All compounds are of analytical grade (BDH, London). Limits for metal detection have previously been established using Techtron. The optimal analytical range has a variable coefficient of 0.87 to 2.20% and is between 0.1 and 0.5 absorbance units. All estimated values are provided as a percentage, even though minerals are reported as milligrams per 100 grams.21
Results
Phytochemical composition of Zanthoxylum acanthopodium DC
The retention factor (Rf) value is the value or size determined based on the position of the stain on the dissolved substance in thin-layer chromatography. The result of the stain produced by the TLC plate is then computed as the Rf.22,23 Table 2 below shows the findings of the phytochemical analysis of Zanthoxylum acanthopodium DC and the resulting Rf values. According to the analyzed metabolites, stains with different colors were produced by identifying phytochemical substances
Table 2. Zanthoxylum acanthopodium DC secondary metabolite screening results.
| Compound | Result | Rf |
|---|---|---|
| Alkaloids | + | Rf1: 0.12 Rf2: 0.20 Rf3: 0.72 |
| Flavonoids | + | Rf1: 0.51 |
| Saponins | + | Rf1: 0.41 Rf2: 0.49 |
| Tannins | + | Rf1: 0.74 Rf2: 0.96 |
| Glycosides | + | Rf1: 0.74 Rf2: 0.96 |
| Steroids/Triterpenoid | + | Rf1: 0.38 Rf2: 0.45 |
Alkaloid compound identification by the TLC method employing chloroform's mobile phase
After spraying with dragendroff, methanol: ammonia (85:15:1) appears as a green stain and yields three stains with Rf values of 0.12, 0.2, and 0.72. The utilized mobile phase has a unique polarity. As a result of chloroform having a higher ratio than methanol and ammonia, the mobile phase is more likely to be non-polar. While substances with higher Rf values are more disseminated in the mobile phase, those with lower Rf values are more distributed in the stationary phase. Because they are more strongly held in the stationary phase compared to compounds with higher Rf values, compounds with lower Rf values are more polar and have more significant distribution coefficients.24,25
Using ethyl acetate to identify flavonoid compounds
One yellow-orange stain with an Rf value of 0.506 was formed by the mobile phase of ethyl acetate: methanol: water (100:13,5:10). Flavonoids are chemicals that are soluble in polar solvents, and when they are separated using mobile phases with semi-polar and polar solubility qualities, the presence of flavonoid molecules in the sample is increasingly evident26
Identification of saponin compounds by TLC using the mobile phase Chloroform: acetic acid; methanol: water (11:6:2:1). The results showed 2 brownish yellow spots with Rf values of 0.41 and 0.49 after being sprayed with methanol: sulfuric acid reagent: vanilla.12
Identification of tannin compounds using the mobile phase of Chloroform-Ethylacetate-n-butanol-Water (11:6:2:1) and 10% FeCl3 stains gave the results of 2 blackish green spots with Rf values of 0.74 and 0.9. This proves that the mobile phase used has the same polarity as tannin, so tannin can be used as an eluent to separate tannin compounds27,28
Identifying glycoside compounds using the TLC method depends on the mobile phase of ethyl acetate: methanol: water (16: 2: 2) produces 2 stains with Rf 0.2 and 0.62 after spraying with 50% sulfuric acid, giving a brown color.12
Identification of steroid/triterpenoid compounds by TLC using the mobile phase n-hexane: ethylacetate (8:2) resulted in 2 stains with a purplish-red color, confirming the presence of steroid/triterpenoid compounds.29
Essential oil analysis
The results of the analysis of essential oils obtained from the gas chromatography–mass spectrometry (GS-MS) chromatogram contained 57 peaks, and six main components were taken based on the highest concentration. The chromatogram results can be seen in Figure 1. The results of the GS-MS analysis showed six main components, namely: geranyl acetate (24.26%, C12H20O2, m/z 196.29), 1-limonene (20.79%, C10H16, m/z 136.24), citronellol (9.72%, C10H20), m/z 156.27), geraniol (7.32%, C10H18O, m/z 154.25), 1,3,6-octatriene (2.67%, C8H12, m/z 108.18), and beta-ocimene (2.92%, C10H16, m/z 136.23). The physiology of the entire plant is so intertwined with essential oil products that it is reliant on metabolic conditions and established developmental differentiation programs of the produced tissues.30 The essential oil content contained in Zanthoxylum acanthopodium DC can be seen in Table 3.
Figure 1. Chromatogram of essential oil of Zanthoxylum acanthopodium DC.
Table 3. Essential oil content in Zanthoxylum acanthopodium DC.
Proximate analysis
Measurements of the ash content, water content, carbs, total fat, and protein were made using proximate analysis. Proximate measures are used to determine the nutritional value of a food product, food ingredient, or food. Analyzing the nutritional value of Zanthoxylum acanthopodium DC will help determine whether it has the potential to be used as a food additive and a therapy. Table 4 displays the findings of the close examination of Zanthoxylum acanthopodium DC. Substance impacts the texture and nutritional formation of the food generated. Bacteria, molds, and yeast can easily thrive in high water content environments and produce changes in the sample's physical or chemical composition.
Mineral analysis
The results of the mineral analysis test contained in the Zanthoxylum acanthopodium DC sample can be seen in Table 5 below. The table shows that the mineral content in the Zanthoxylum acanthopodium DC sample is Lead (Pb), Cadmium (Cd), Arsenic (As), Mercury (Hg), Manganese (Mn), Potassium (K), Calcium (Ca), Magnesium (Mg), Iron (Fe) and Sodium (Na).
Table 5. Zanthoxylum acanthopodium DC mineral analysis data.
Discussion
The fact that Zanthoxylum acanthopodium DC was found to contain phytochemicals such alkaloids, tannins, steroids/triterpenoids, flavonoids, saponins, and glycosides supports its usage as a medicinal plant. These secondary plant metabolites' therapeutic capabilities have also been described by a number of writers.31–34 There is evidence that alkaloids can stimulate the central nervous system.35 The antiviral, antifungal, antibacterial, and anticancer effects of tannins are well documented. They offer a variety of pharmacological qualities that have been linked to wound healing, including anti-inflammatory, analgesic, and antioxidant activities.34,36 The antibacterial properties of tannins are applied through breakdown of membranes, protein binding, adhesion, and inhibition of enzymes.37 According to reports, detected steroids and triterpenoids exhibit anti-bacterial, antiviral, antimalarial, and anti-cholesterol synthesis inhibiting activities.35–37 Antibacterial activities of steroids and triterpenoids have been documented.38,39 They have long been used medicinally, and one of its common biologic characteristics is cytotoxicity.40 Flavonoid content in Zanthoxylum acanthopodium DC has the potential as an antioxidant. Flavonoid can act as antioxidants with the mechanism of chelating or scavenging.41,42 Saponins also detected were known to have the properties of precipitating and coagulating red blood cells.41 Some characteristics of saponins include foam formation in aqueous solutions, hemolytic activity, cholesterol-binding properties and bitterness.43,44 Sodipo et al. (2000) reported that saponins lower cholesterol levels, act as immune boosters and are anti-carcinogenic.43 However, it was also reported that high levels of saponins could cause gastroenteritis.45 There is a low amount of saponins in Zanthoxylum acanthopodium DC, which confirms that its consumption will not harm the human body. In addition, according to many reports, Zanthoxylum acanthopodium DC fruits contain glycosides known to lower blood pressure.46
In addition to the metabolite content, the results of this study also showed that Zanthoxylum acanthopodium DC contains essential oils. Essential oils are part of a group of volatile compounds and give off a characteristic odor. The most significant ingredients contained in Zanthoxylum acanthopodium DC are essential oils of geranyl acetate (24.26%) and 1-limonene (20.79%). The compounds -myrcene, (z)-β-ocimene, linalool, -citronellol, neral, geraniol, geranyl acetate and sesquiterpenoids contribute to the distinctive aroma of Zanthoxylum acanthopodium DC. Zanthoxylum acanthopodium DC essential oil has potential in vitro cytotoxic effects on T47D breast cancer cells.9,10 Essential oil compounds in Zanthoxylum acanthopodium DC can also inhibit the growth of bacteria.47
The results of the proximate analysis also show that Zanthoxylum acanthopodium DC has a sample moisture content (of 6.35%). The water content value of this sample still meets the Standard National Indonesia (SNI) No 01-2891-1992 criteria even though it is lower than the range of 15-24% allowed by SNI.48,49 The ash content of the sample is 6.19%, a measure used to determine the number of minerals present in the sample.50,51 Ash content is a metric that indicates the purity of a sample and is affected by its composition and mineral content. The total fat value is 2.46%, the protein value is 16.2%, and the carbohydrate value is 35.4%. A good source of protein was defined as having a protein content that exceeded 12% of its caloric value, and a crude fiber content of 33.4% sample was high enough to meet the recommended daily allowance of fiber.52,53 Carbohydrates and crude fiber, such as lignin and pectin, are the two substances in the samples of Zanthoxylum acanthopodium DC in the highest concentrations. Total fat is the least abundant element in Zanthoxylum acanthopodium DC.
Every herbal plant provides health benefits because it has nutritional value. Zanthoxylum acanthopodium DC is detected to contain minerals such as Lead (Pb), Cadmium (Cd), Arsenic (As), Mercury (Hg), Manganese (Mn), Potassium (K), Calcium (Ca), Magnesium (Mg), Iron (Fe) and Sodium (Na). Minerals are essential for the proper functioning of tissues and act as secondary messengers in several biochemical cascade mechanisms.54,55 For example, calcium is an important mineral content found in plants because of its many roles in the life of living things. The content of magnesium plays a role in the activity of manganese (Mn), which is considered an antioxidant nutrient and breaks down bile, thereby reducing the risk of cholesterol. Lack of calcium in the body can cause several disorders, for example, inhibiting growth.1,10,53 Potassium in Zanthoxylum acanthopodium DC plays an essential role in diuretics and regulates cardiac function, water, and ion balance in blood and tissues.56,57 Calcium content is well known for the growth and maintenance of bones, teeth and muscles.58,59 In addition, calcium is helpful in the formation of blood and intracellular and extracellular fluids inside and outside the body's cells.60 The iron content is also essential. Iron plays a vital role in erythropoiesis and many intracellular oxygen transport reactions. It facilitates the oxidation of carbohydrates, proteins and fats.61–63 Sodium and potassium are needed to maintain the osmotic balance of body fluids and body pH, regulate muscle and nerve irritability and control glucose uptake.63
Conclusions
The study's findings by Zanthoxylum acanthopodium DC demonstrated that the sample's phytochemical, mineral, and essential oil content is strongly associated with its potential to be developed as food and medicine. Every plant must include the biological substances required to serve as metabolites for creating novel medications and foods, so that more comprehensive studies on the effectiveness and compound interactions of these compounds' contents might be developed.
Download
Export To
metrics
| Views | Downloads | |
|---|---|---|
| F1000Research | - | - |
|
PubMed Central
Data from PMC are received and updated monthly.
|
- | - |
Citations
CITE
how to cite this article
Satria D, Dalimunthe A, Pertiwi D et al. Phytochemicals, proximate composition, minerals and volatile oil analysis of Zanthoxylum acanthopodium DC. fruits [version 1; peer review: 1 not approved]. F1000Research 2023, 12:227 (https://doi.org/10.12688/f1000research.128941.1)
NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article.
track
receive updates on this article
Track an article to receive email alerts on any updates to this article.
Open Peer Review
Current Reviewer Status:
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 1
VERSION 1
PUBLISHED 01 Mar 2023
Views
6
How to cite this report:
Alam F. Reviewer Report For: Phytochemicals, proximate composition, minerals and volatile oil analysis of Zanthoxylum acanthopodium DC. fruits [version 1; peer review: 1 not approved]. F1000Research 2023, 12:227 (https://doi.org/10.5256/f1000research.141581.r216956)
The direct URL for this report is:
https://f1000research.com/articles/12-227/v1#referee-response-216956
https://f1000research.com/articles/12-227/v1#referee-response-216956
NOTE: it is important to ensure the information in square brackets after the title is included in this citation.
Reviewer Report 02 Nov 2023
Fiaz Alam,
Department of Pharmacy, Comsats University Islamabad, Islamabad, Pakistan
Not Approved
VIEWS 6
Thanks for inviting me to review the manuscript “Phytochemicals, proximate composition, minerals and volatile oil analysis of Zanthoxylum acanthopodium DC. Fruits”
The idea of the manuscript is fine but its not suitable to be indexed in the ... Continue reading
The idea of the manuscript is fine but its not suitable to be indexed in the ... Continue reading
CITE
HOW TO CITE THIS REPORT
Alam F. Reviewer Report For: Phytochemicals, proximate composition, minerals and volatile oil analysis of Zanthoxylum acanthopodium DC. fruits [version 1; peer review: 1 not approved]. F1000Research 2023, 12:227 (https://doi.org/10.5256/f1000research.141581.r216956)
The direct URL for this report is:
https://f1000research.com/articles/12-227/v1#referee-response-216956
https://f1000research.com/articles/12-227/v1#referee-response-216956
NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article.
Comments on this article Comments (0)