Open access peer-reviewed chapter

Phytochemical Profiling of Soybean (Glycine max (L.) Merr.) Genotypes Using GC-MS Analysis

Written By

Salem Alghamdi, Hussein Migdadi, Muhammad Khan, Ehab H. El- Harty, Megahed Ammar, Muhammad Farooq and Muhammad Afzal

Submitted: 25 March 2018 Reviewed: 27 April 2018 Published: 07 November 2018

DOI: 10.5772/intechopen.78035

From the Edited Volume

Phytochemicals - Source of Antioxidants and Role in Disease Prevention

Edited by Toshiki Asao and Md Asaduzzaman

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Twenty-four soybean genotypes collected from different regions and origin were evaluated for their quality performance to explore their nutritional and medicinal values. The proximate compositions showed considerable variations among soybean genotypes. The USA genotypes recorded the highest values for protein (43.1 g/100 g), total fat (23.61 g/100 g), phenolic content and flavonoids (1.77 and 2.13 mg/g). Using GC-MS analyses of methanolic extracts, a total of 88 compounds were identified in the genotypes and were classified to: 19 heterocyclic compounds, 13 compounds for ketones and esters, 9 for phenolic compound, 7 compounds for carboxylic acids and sugar moiety, 5 compounds for aldehydes and alcohols, 4 ether compounds, 3 amide, 2 alkanes and one alkene and one fatty acid ester. Indonesian genotypes recorded the highest number of phenolic and the Australian genotype A-1 had the maximum number of esters. Genotypes showed high levels of proximate compositions and pharmaceutical components, offering potential candidates for improving those traits in adapted genotypes through breeding program, as well as serving as a good source of mass production of pharmaceutical and medicinal components either through classical or in vitro production. Furthermore, platform was set for isolating and understanding the characteristics of each compound for it pharmacological properties.


  • soybean
  • phenolic compounds
  • GC-MS
  • flavonoids
  • nutritional value

1. Introduction

Soybean (Glycine max (L.) Merr) considered among ancient cultivated crops, it was domesticated in the 11th century BC around Northeast of China. It is one of the most widely grown leguminous crops in the world. Its cultivated area was recorded in 95 countries more than 121 million hectare that produced 335 million tons of dry seeds [1] (FAOSTAT, 2016). Soybean had a wide variability, the USDA alone maintains more than 15 thousand soybean accession grouped into 13 maturity classes including both determinate and indeterminate soybean. Early maturing groups are adapted to short summer growing seasons in North USA and Canada while late maturity groups are adapted to southern or coastal plain counties [2]. Soybean occupies an advanced position among agricultural crops, being the most important source of proteins and vegetable oils [3]. Its seeds provided abundant and high quality protein and oil for human diet and animal feed. Its seeds contain more than 36% protein, 30% carbohydrates in addition to fiber, vitamins, and minerals [3]. It also contains about 20% oil, which makes soybean one of the most important edible oil crops. Soy oil has used as binding additives in manufacturing of papers, inks, paints, varnishes, cosmetics, and plastics. It was used also in production of farming pesticides and pharmaceuticals products [4]. Nowadays, biodiesel utilizing soy oil become a new industrial renewable sources of energy. Additionally, soybean as a nitrogen-fixing legume crop helps in reducing the chemical source of nitrogen fertilizers production [4].

Furthermore, tofu, soy milk, soy sauce, miso, etc., have been developed for human consumption, while soya meal (oil extraction by-product) is used as a nutritious animal feed [5]. Moreover, soybean is now regarded as a model legume crop owing to the availability of genome sequence information [6]. Keeping in mind its vast uses, there is huge number of justifications for crop improvement programs throughout the world. Having 53% global production share of all oilseed crops, USA, China, Brazil, Argentina and India gave soybean much attention in the agricultural production systems. Yield and total production of soybean increased over the last two decades due to genetic improvement of this crop [7].

In comparison with conventional legume and animal feed sources, soybean is considered one of the cheapest food resources with medicinal properties due to their highest protein content and no cholesterol due to its contents of Genistein, photochemical and isoflavones [8]. It can help in disease fighting due to its pharmacological properties and its phytochemicals constitutes, including antioxidant, estrogenic, antidiabetic, anti-hypercholesterolemic, anti-hyperlipidemic, anti-obesity, antihypertensive, anticancer, anti-mutagenic, hepatoprotective, anti-osteoporotic, antiviral, bifidogenic, anti-inflammatory, immunomodulatory, neuroprotective, wound healing, antimicrobial, goitrogenic anti-skin aging, anti-photoaging activity and the effects of anti-nutritional factors [3]. A 111 volatile compounds in fermented soybean curds were reported by Chung [9] and an 83 in commercial plain sufu [10]. Messina [11] reported that the presence of isoflavones in soybean is behind the pharmacological attributes of this crop. Chemical composition included Phenolic acids, flavonoids, isoflavones, saponins, phytosterols and sphingolipids were recorded in soybean [12, 13, 14]. Due to importance of this crop and its products, this study was amid at estimating the most active constitutes of 24 soybean genotypes including total phenolic, flavonoid and protein content and phytochemicals using GC-MS.


2. Materials and methods

2.1. Plant materials

Twenty-four soybean genotypes were grown in Dirab Agriculture Research Station, King Saud University, Riyadh, Kingdom of Saudi Arabia (24_25049.200 N 46_22012.500E) on August, 2014 and were collected from nine countries (Argentina, Australia, China, Egypt, India, Indonesia, USA, and Pakistan). The name and geographical origin of these genotypes are presented in the Table 1.

Entry no. Genotype name Source/origin Entry no. Genotype name Source/origin
1 Admaril Pakistan 13 Giza 111 Egypt
2 Romal-1 Pakistan 14 Clark USA
3 NARC-2 Pakistan 15 3803 Syria
4 Williams 82 USA 15 A-1 Australia
5 X 32 Egypt 17 Ijen Indonesia
6 Giza 22 Egypt 18 Indo-black Indonesia
7 Giza 21 Egypt 29 Indo-I Indonesia
8 X2 L 12 Egypt 20 Indo-II Indonesia
9 Giza 83 Egypt 21 USA-1 USA
10 Crawford USA 22 Indian India
11 Giza 35 Egypt 23 Chinese China
12 X 30 Egypt 24 Argentinian Argentina

Table 1.

Name and source of the 24 soybean genotypes invistigated in the study.

2.2. Chemical analysis

2.2.1. Proximate composition

Triplicate sample is used to determine the proximate analysis of soybean genotypes for crude proteins, moisture, total ash, fat and carbohydrate by using the methods described in AOAC, [15]. Protein content was estimated using Kjeldahl method with titration and percent nitrogen was determined using [16] equation.

2.2.2. Antioxidants determination

Soybean samples approximately (1 g) were powdered and homogenized in 10 ml 80% methanol. The mixture was shaken at 300 rpm at room temperature for 3 h. Then the extract was centrifuged for 10 min at 3000 rpm and upper aqueous phase were transferred to new Eppndorf tubes. Moreover, the residues were again extracted with 5 ml 80% methanol overnight. The extraction was performed in three replicates, later on extracts combined and stored in dark at 4°C. The Folin-Ciocalteu reagent was used to determine the total phenolic compounds from the extracts using gallic acid calibration curve as standard. The total phenolics were expressed as mg/g gallic acid equivalents (GAE). An extract was aliquot (50 μl) and mixed with Folin-Ciocalteu reagent of 250 μl and 7.5% sodium carbonate of 750 μl. The volume was increased to 5 ml with water and sample was incubated for 2 h. The absorbance was measured at 765 nm against distilled water as blank. The flavonoid determination was measure by aluminum chloride method with the help of Quercetin equivalent as standard. An aliquot of extract (250 μl) was mixed with ddH2O and 5% NaNO2 (15:1, v/v). After 6 min, 150 μl of 10% AlCl3 was added to the mixture. A 500 μl of 1 M NaOH was added to the mixture at the 5th min, and volume made up to 2.5 ml with distills water and the absorbance was measured spectrophotometrically at 410 nm.

2.2.3. Gas chromatography-mass spectroscopy

The GC-MS analysis of fractions were performed using a TSQTM 8000 Evo Triple Quadrupole GC-MS/MS (Thermo Fisher Scientific) equipped with an Elite-5 capillary column (length 30 nm and inner diameter 0.25 mm and film thickness 0.25 μm) and mass detector was operated in electron impact (EI) mode with full scan (50–550 amu). Helium was used as the carrier gas at constant flow rate 1 mL/min and an injection volume of 1 μL. The oven injector temperature was programmed from 50°C with an increase of 8°C/min to 200°C, then 7°C/min to 290°C/min. The results were compared using the database of National Institute Standard and Technology (NIST).

2.3. Data analysis

The data were subjected to descriptive statistics (mean, standard deviation, coefficient of variability, minimum and maximum values) and principal component analysis (PCA) using statistical software Past3 program [17].


3. Results and discussion

3.1. Proximate analysis

The proximate analysis values of 24 soybean genotypes (crude protein, ash fat, carbohydrate, and moisture contents) values and total phenolic and flavonoid contents are shown in Table 2, and the detailed proximate analysis estimates are presented in Table S1. The minimum crude protein value was recorded for Argentinian (35.63%), while maximum recorded for Clark genotypes (43.13%). The genotypes, i.e., Clark, Indo-1, Indo-black, Ijen, Romal-1, X 30 and 3803 recorded higher than 40% crude protein. The significant variations for crude proteins among genotypes were recorded and that might observed due to differences in genetic background and/or origin. The higher protein content in the genotypes is also reported previously which ranged from 43 to 45% [18]. These results are also in line with Zarkadas et al. [19, 20] who reported crude protein contents in soybean ranging from 33.67 to 42.11%. The minimum moisture contents were recorded in Giza 83 (3.08%) while maximum was recorded for Indo-1 (5.88%) with an average (4.90%) mean value showing non-significant difference. Ash contents ranged from 4.55 to 6.28% with an average of 5.44%. The maximum was recorded for Giza 111 (6.28%) genotype while Romal-1 genotype had the lowest (4.55%) of ash contents. The moisture and ash contents values were recorded lower than that reported by [21]. Total fat ranged from 16.92 to 22.94% with a mean value of 21.16%. The genotype Indo-black contained the lowest while the genotype 3803 recorded the highest content. Soybean is considered about 47% of its energy value in fat content [22, 23] which is compared to other legumes. Our results regarding total fat were in line with that of [24] who reported that that total fat value ranged 18 and 22 g/100 g in soybean genotypes. The minimum carbohydrate content in Clark (26.11%) while maximum in Argentinian (33.18%), with an average (29.48%) was recorded among soybean genotypes.

Crude protein (g/100 g) Moisture (g/100 g) Ash (g/100 g) Total fat (g/100 g) Carbohydrate (g/100 g) Total phenolic content (TPC) Total flavonoid content (TFC)
N 24 24 24 24 24 24 24
Min 35.63 3.08 4.55 16.92 26.11 1.15 0.68
Max 43.13 5.88 6.28 23.61 33.18 1.77 2.13
Mean 39.02 4.90 5.44 21.16 29.48 1.45 1.24
Stand. dev. 2.09 0.65 0.33 1.41 1.86 0.16 0.36
Coeff. Var. 5.35 13.26 6.11 6.68 6.30 11.58 29.32

Table 2.

Descriptive statistics of chemical composition in 24 soybean genotypes.

3.2. Flavonoid and phenolic contents

Flavonoid and phenolic compounds are the important phytochemicals and natural antioxidants founds in fruits, vegetable and cereals grains. It serves as multiple biological functions, i.e., defense against cardiovascular disease, cancer and aging [25]. The results regarding total phenolic and flavonoids contents for 24 soybean genotypes are presented in Table S1, and significant differences were recorded for all soybean genotypes. The seed extracted results indicated that the maximum phenolic contents was recorded in Romal-1 (1.7 mg/g) while minimum in Giza 111 (1.15 mg/g) with an average 1.45 GAE/g mg/g (Table 2). However, total flavonoid content ranged 0.68 to 2.13 mg QE/g (Table 2). Phenolic content is strongly linked with antioxidant capacity [26, 27] and can contribute towards antioxidants activities [28]. The use and demands of phenolic are increasing rapidly in food industry to enhance nutritional value and quality of food [29].

3.3. GC-MS analysis

Methanolic extracts of 24 soybean genotypes using GC-MS analysis were used to identify a large number of phytochemical. Based on peak area, retention time and molecular formula, about 88 compounds were recognized. A large number of bioactive phytochemicals including flavonoids, phenolic acids, saponins, isoflavones, sphingolipids and phytosterols were also reported previously for soybean [12, 13, 14]. The carbamide was the first compound that identified at 3.67 min retention time, whereas, last compound identified at 48.53 min retention time was methyl 10 Trans, 12-cisoctadecadienoate recognized at 48.53 min retention time (Table S2). A wide difference was recorded for composition of phytochemical in 24 soybean genotypes. The phytocompounds and their biological activities in soybean genotypes were presented in Table 3. The phytocompounds of the studied soybean genotypes divided into different groups (Figure 1). The resulted 88 compounds were categorized into heterocyclic compounds (19), aldehydes (5), alcohols (5), esters (13), amide (3), sugar moiety (7), ether (4), phenolic compound (9), carboxylic acids (7), ketones (13), alkanes (2), one fatty acid ester and one Alkene. A typical chromatogram of one soybean genotype was shown in Figure 2. The GC-MS analyses showed that the methanolic extract is largely composed of heterocyclic compound, ester and phenolic compound. Hexadecanoic acid, methyl ester, 2,6-dimethoxy, 3,5-dimethoxyacetophenone, 2-methoxy-4-vinylphenol, phenol and 1,2-cyclopentanedione were noticed in most of the genotypes. These phytochemicals are involved in various pharmacological actions, i.e., antioxidants and antimicrobial activities [30]. These chemicals are also active in many biological activities that were listed (Table S2). Phytochemicals also possess antioxidant activities, anti-cancer, anticarcinogenic, antibacterial, antiviral, or anti-inflammatory activities and play an important role for plant metabolism [30, 31]. The five compounds belong to aldehyde group (benzeneacetaldehyde, 3,4-dimethylbenzaldehyde, methoxy-propanal, p-hydroxyphenyl, glyoxal and propanal, 2-(benzoyloxy)-, benzeneacetaldehyde), were detected in 10 genotypes (Table S2). Admiral and Williams 82 contains 3-methoxy-propanal while indo-black, Indo-1 and Indo-II contains 3,4-dimethylbenzaldehyde, whereas Giza 35 and X30 contains p-hydroxyphenyl) glyoxal and propanal, 2-benzoyloxy, respectively. The highest number of aldehyde compounds is present in William 82 genotype (2). It is also reported that; aldehyde possess powerful antimicrobial activity due to their highly electronegative arrangement of conjugated group C=C double bond [32], as the electronegativity increase, antimicrobial activity also increases in those genotypes [33, 34]. These compounds react with vital nitrogen components such as protein and nucleic acid, consequently inhibit microorganism. Thirteen ketone related compounds were identified, i.e., 1-(dimethylamino)-, 1,2-propanone, 1,2-cyclopentanedione and 6-Oxa-bicyclo [3.1.0] hexan-3-one, 2,4-dihydroxy-2,5-dimethyl-3(2H)-furan-3-one,2-acetyl-2,3,5,6-tetrahydro-1,4-thiazine,butyrolactone, 2,5-Dimethyl-4-hydroxy-3(2H)-furanone, 5-hepten-3-one, 5-methyl-, dihydroxyacetone, 2-pyrrolidinone, 1-methyl, 2,4,6,-cycloheptatrien-1-one,4-methyl-, 3,5-dimethoxyacetophenone. The indo-11 and 3803 genotypes recorded highest ketonic compounds (8) followed by present in Giza 35 and USA-1 genotypes that contained 6ketonic group each. Ketones might be formed by beta-oxidation of fatty acid and have some important flavor compounds [35]. During fatty acid metabolism, many volatile compounds are also formed, producing alcohols, acids and esters. Many alcoholic compounds are derived from bioremediation of unsaturated fatty acids and are prerequisite for the formation of long chain esters. These identified compounds in soybean genotypes are 4-methyl-2-haptanol, 1,2,3-propanetriol, isosorbide (D-glucitol, 1,4,3,6-dianhydro), 1-undecanol alcohol, and 1,3-dioxolane-4-methanol (glycerol formal). 4-Methyl-2-haptanol was present in Genotype Giza 35 while 1,2,3-propanetriol was present in nine genotypes and isosorbide was detected in three soybean genotypes. The highest alcoholic compounds (3) were detected in Clark genotype as compared to other genotypes. Alcohols also possess antibacterial activity against vegetative cell. Glycerol and derivatives also show bacterial inhibiting effect [36]. The following seven carboxylic acids namely acetic acid, 2-pyridinecarboxylic acid (also called picolinic acid), 2,2-[oxybis(2,1-ethanediyloxy)]bis, butanoic acid, 4-hydroxy-, propyl-(also called 2-propylmalonic acid), propanedioic acid, benzoic acid, butanoic acid, 4,4-dithiobis[2-amino-,[S-(R/,R/)] were detected (Table S2). Five genotypes were having acetic acid and 2-pyridinecarboxylic acid was present in five genotypes. Three genotypes have butanoic acid and 4-hydroxy-was appeared in three genotypes while one genotype has benzoic acid. Giza 35, X30, Argentinian and Chinese compassed the maximum numbers of carboxylic acids compounds. Thirteen esters were identified. The butyrolactone, acetic acid, 2-(dimethylamino)ethyl ester, formic acid, 3-methylbut-2-yl ester, pentanoic acid, 2-isopropoxyphenyl ester, phthalic acid, hex-3-yl-isobutyl ester, hexadecanoic acid, methyl ester, phthalic acid, butyl undecyl ester, 5,8,11-heptadecatriynoic acid methyl ester, methyl 10-trans, 12-cis-octadecadienoate, 9,12-octadecadienoic acid(Z,Z)-methyl ester, benzoic acid, 4-ethoxy-, ethyl ester, 1,2-benzenedicarboxylic acid, dibutyl ester, and pentanoic acid, 2,2-4-trimethyl-3-carboxyisopropyl, isobutyl, ester were identified. The genotype A-1 had maximum six esters compounds followed by others genotypes (Giza 83, Romal-1, Clark, Argentinian and 3803) having five (5) esters compounds. Hexadecanoic acid ethyl ester shows antioxidant, nematicidal activities and hypocholesterolemic [37]. Regarding phenolic compound, a total of nine compounds were identified. 1,2-benzenediol,3-methoxy-, 5-tert-butyl-1,2,3-benzenetriol, phenol, 4-ethenyl-, acetate, phenol, 2,6-dimethoxy-, 2-methoxy-4-vinylphenol, phenol,2,6-bis(1,1-dimethylethyl)-4-methy l-, phenol, 2,4-bis(1,1-dimethylethyl)-, and phenol, 2-methoxy. The genotypes Indo-1 and Ijen and recorded the highest number of phenolic compounds which is five while the genotypes Clark, NARC-2, Giza 35, USA-1 and Indo-11 contained the four (4) phenolic compounds each. The plant phenolics compounds are of great interest to human due to their anti-oxidative and possible anticarcinogenic activities. The dietary phenolics are considered anti-carcinogens because of antioxidants, but there is no clear proof supporting this supposition [38]. Phenolic may inhibit carcinogenesis by interfering the molecular events in initiation, promotion, and progression stages. Isoflavones and lignans from soybean may distract tumor formation by mediating estrogen-related activities and also modulate the growth of benign and malignant prostatic epithelial cells in vitro [39]. The following sugar moiety, L-galactose, 6-deoxy-, 3,4-0-isopropylidene-d-galactose, a-methyl-D-m annopyranoside, 3-O-methyl-d-glucose a-D-galactopyranoside, methyl were appeared among soybean studied genotypes. The relatively notable amounts of heterocyclic compounds were identified including 3,5-dihydroxy-6-methyl-2, 2,6-diisoprpylnapthalene, 4H-pyran-4-one, 3-dihydro-4Hpyran-4-one, 3-hydroxy-2-methyl-, pyrazine, ethyl-, oxirane, 2-ethyl-2-methyl, 1H-indazole, 4,5,6,7-tetrahydro, N-aminomorpholine, and benzofuran, 2,3-dihydro. The genotype X30 had four sugars compounds while genotypes USA-1, Indo-1, and Indo-11 had three sugars compounds each. Benzofurans are considered to possess anti-oxidant, antimicrobial effect and anti-inflammatory [40]. The compounds detected in this study have reported to have potentials as therapeutic agents, antioxidant, antimicrobial, and anti-inflammatory compounds and demonstrating that different compounds can exhibit similar activity and this might be due to presence of similar functional groups (Table S2). Antioxidant properties of soybean extract could be the basis for the presence of various antioxidant and anti-inflammatory compounds.

Compound Other names Nature Activity RT MW
22 2H-1-Benzopyran,3,5,6,8a-tetrahydro-2,5,5,8a-tetramethyl-,(2S-cis)- Edulan II Heterocyclic compound 7.58 192
27 1,2-Cyclopentanedione Ketone Antioxidant 7.98 98
28 Pyran-4-Carboxylic acid, 4-(4-methoxyphenyl)-tetrahydro- Heterocyclic compound 8.02 236
34 2,4-Dihydroxy-2,5-dimethyl-3(2H)-furan-3-one Ketone 9.74 144
36 2H-Pyran-2,6(3H)-dione Glutaconic anhydride Heterocyclic compound 10.75 112
39 2-Pyrrolidinone, 1-methyl M-Pyrol Ketone 11.86 99
42 2,5-Dimethyl-4-hydroxy-3(2H)-furanone Ketone 12.28 128
44 Phenol, 2-methoxy- Phenolic compound Antimicrobial, antioxidant, anti-inflammatory, analgesic 13.83 124
49 4H-Pyran-4-one,3-hydroxy-2-methyl- Maltol Heterocyclic compound Flavor enhancer 14.78 126
50 5-Hepten-3-one, 5-methyl- Ketone compound 15.09 126
52 3,5-Dihydroxy-6-methyl-2,3-dihydro-4H-pyran-4-one Heterocyclic compound Antimicrobial, anti-inflammatory 16.61 144
57 Phenol, 4-ethenyl-, acetate 4-Vinylphenyl acetate Phenolic compound Antimicrobial, antioxidant, anti-inflammatory 19.29 162
60 Benzofuran, 2,3-dihydro Coumaran Heterocyclic compound Antihelminthic, anti-inflammatory, antidiarrheal 20.16 120
62 Benzeneacetaldehyde, 3-methyl m-Tolualdehyde Aldehyde Antimicrobial 20.34 120
61 1,2-Benzenediol,3-methoxy- Pyrocatechol, 3-methoxy Phenolic compound Antioxidant 21.01 140
64 2-Methoxy-4-vinylphenol Phenol, 4-ethenyl-2-methoxy- Phenolic compound Antimicrobial, antioxidant, anti-inflammatory 23.35 150
68 Phenol, 2,6-dimethoxy- Pyrogallol 1,3-dimethyl ether Phenolic compound Antimicrobial, antioxidant, anti-inflammatory 24.99 154
70 Phenol,2,6-bis(1,1-dimethylethyl)-4-methyl- Butylated hydroxytoluene Phenolic compound Antimicrobial, antioxidant, anti-inflammatory, analgesic 30.99 220
71 Phenol, 2,4-bis(1,1-dimethylethyl)- Phenol, 2,4-di-tert-butyl- Phenolic compound Antimicrobial, antioxidant, anti-inflammatory 31.19 206
72 5-tert-Butyl-1,2,3-benzenetriol 5-Tert-butylpyrogallol Phenolic compound Antioxidant, antiseptic, antibacterial, anti-dermatitic fungicide, pesticide 31.91 182
76 3,5-Dimethoxyacetophenone Ketone Antioxidant 33.65 180
85 Hexadecanoic acid, methyl ester Palmitic acid, methyl ester Ester Antioxidant, flavor, hypocholesterolemic, nematicide 46.13 270

Table 3.

List of important phytocompounds identified in the methanolic seed extract of soybean genotypes by GC-MS.

Figure 1.

Pie diagram showing the percentage of phytochemical groups identified in 24 soybean genotypes.

Figure 2.

A typical GC-MS profile of seeds of soybean genotype.

3.4. Principal component analysis (PCA)

The first three principal components explained 78.64% of total variations among genotypes (Table 4 and Figure 3). The first component described 59.65% of total variation, and positively correlated with phytochemical classes of ether, alcohol, sugar moiety ketone and phenolic compounds. Genotypes Ijen, Clark, A-1, USA-1, Indo-II, 3803, X 30, Giza 35, Indo-black and Indo-I showed the most variability according to these components and can be selected for these classes. PC2 illustrated 10.63% of the total variance, and the amide, sugar moiety, ether, alkane, ketone and carboxylic acid positively correlated with this component. The genotypes showed most variability were Giza 111, Giza 35, X 30, X 32, Indo-II and 3803. Alkane, Aldehyde, Carboxylic acid and Phenolic compound were positively correlated with the third component. The genotypes Giza 35, X 32 showed most variability based on this component. In this study, genotypes Giza 35, X 30, Indo-II and genotype 3803 showed positive loading in at least two out of the three PCs, which can be utilized in breeding for ceratin class of phytochemical. Utilizing PCA effectively reduces the number of variables needed to classify cultivars and permitted soybean researchers to more easily develop significant relationships between important soybean characteristics. Soybean cultivars have been classified using (PCA) of the fatty acid data [41]. The first four principal components generated in total 81.49% of the variance, where PC1 positively correlated with oleic, linoleic, and gondoic acids, PC2 with stearic, linolenic and arachidic acids, PC3 behenic and lignoceric acids, and PC4 by palmitic acid. Moreover, due to the ability of PCA to manage and interpret large data sets, it has been used in studying relationships that exist in fatty acid characterization [42]. Although soybean oil has been included in some chemometric studies comparing vegetable oils, soybean cultivars have yet to be extensively classified using multivariate techniques [43, 44].

PC 1 PC 2 PC 3
Eigen values 0.17 0.03 0.02
Percent of variance 59.65 10.63 8.36
Cumulative percentage 59.65 70.28 78.64
Alcohol 0.42 0.11 −0.12
Aldehyde −0.14 0.00 0.24
Alkane −0.01 0.29 0.75
Amide −0.59 0.67 −0.28
Sugar moiety 0.39 0.44 −0.06
Carboxylic acid 0.03 0.06 0.31
Ester 0.12 −0.25 −0.26
Ether 0.45 0.34 0.02
Heterocyclic compound 0.05 0.14 −0.16
Ketone 0.27 0.21 −0.18
Phenolic compound 0.11 −0.09 0.24

Table 4.

Eigen values and proportion of the variance explained for the three principal components of the 24 soybean genotypes based on phytochemical components.

Figure 3.

Two-dimensional biplot ordination of 24 soybean genotypes on principal component axes according to 11 phytochemical classes.


4. Conclusion

The results revealed that soybean genotypes cover variable patterns of total proteins flavonoids, phenolic and various bioactive volatile compounds. The mass spectrometry analysis results demonstrated that, majority of soybean genotypes are a source bioactive compounds with antioxidant, anti-inflammatory, antimicrobial and other functions. 2-Methoxy-4-vinylphenol, phenol, 2,6-dimethoxy-, 3,5-dimethoxyacetophenone, hexadecanoic acid methyl ester, 1,2-cyclopentanedione, and 3,5-dihy droxy-6-methyl-2,3-dihydro-4H-pyran-4-one were present in majority of genotypes. However, the genotypes Ijen and Indo-1 contributed more phenolic compound than others genotype. Genotype A-1 has the maximum compound in esters compounds. The genotypes Indo-11 and 3803 contribute maximum ketone compounds while Giza 111 contributes more in heterocyclic compounds. Some genotypes may have good therapeutic potential and could be served as a potential source in drug wdevelopment as a health supplement. This study also provides a platform for isolating and understanding the properties of each compound for it pharmacological properties.



The authors of this project in number AT-34-58 are highly appreciated the encouragement and the assistances provided by the King Abdulaziz City for Science and Technology.


Conflict of interest

The authors have declared that no conflict of interest exists.


Genotype name Crude protein (g/100 g) Moisture (g/100 g) Ash (g/100 g) Total fat (g/100 g) Carbohydrate (g/100 g) Total phenolic content mg/g Total flavonoid content mg/g
Admaril 37.84 4.56 5.27 21.65 30.68 1.30 0.975
Romal-1 40.93 4.79 4.55 20.35 29.38 1.75 1
NARC-2 38.01 4.84 5.79 21.16 30.2 1.50 1.25
Williams 82 38.23 4.97 5.65 22.79 28.36 1.42 1.05
X 32 39.8 4.31 5.54 21.04 29.31 1.25 0.875
Holladay 37.04 4.35 5.55 23.61 29.45 1.35 0.75
Giza 22 39.82 4.45 5.55 21.91 28.27 1.37 0.675
Giza 21 39.84 4.4 5.56 21.72 28.48 1.40 0.8
X2 L 12 38.26 4.26 5.29 21.96 30.23 1.42 1.2
Giza 83 38.29 3.08 5.39 21.42 31.82 1.38 0.925
Crawford 39.43 4.84 5.58 22.38 27.77 1.30 1.125
Giza 35 38.8 3.77 5.49 21.78 30.16 1.32 1.025
X 30 40.05 4.99 5.64 21.78 27.54 1.70 1.0375
Giza 111 36.89 5.34 6.28 22.07 29.42 1.15 1.75
Clark 43.13 5.41 5.77 19.58 26.11 1.35 1.375
3803 40 5.12 5.45 22.94 26.49 1.33 1.625
A − 1 39.01 5.19 4.8 20.69 30.31 1.37 1.45
Ijen 41.7 5.54 5.54 18.66 28.56 1.65 1.25
Indo-black 42.71 5.88 5.36 16.92 29.13 1.65 1.025
Indo-I 42.74 5.88 5.7 19.33 26.35 1.62 1.775
Indo-II 37.87 5.51 5.14 21.17 30.31 1.32 1.375
USA-1 36.89 5.34 5.24 21.34 31.19 1.77 2.125
Indian 36.59 5.43 5.25 20.88 31.85 1.65 1.7625
Chinese 35.98 5.19 5.26 21.06 32.51 1.50 1.35
Argentinian 35.63 5.12 5.3 20.77 33.18 1.37 1.325

Table S1.

Proximate analysis, total phenolic and flavonoid in the seeds of 24 soybean genotypes seeds (on a dry weight basis).

Sr. no Compound Other name Nature Activity RT MW
1 Carbamide Urea Amide 3.67 60
2 Propanal, 3-methoxy 3-Methoxy-propanal Aldehyde Antibacterial 3.75 88
3 n-Hexane Alkane Antibacterial 3.8 86
4 Acetamide, oxime Amide Antimicribial 3.86 74
5 1,2-Naphthalenedione, 4 chloro Heterocyclic compound 3.92 192
6 1,3-Dioxolane-4-methanol Glycerol formal Alcohol 3.93 104
7 1-Monolinoleoyglycerol trimethylsilyl ether Ether 4.02 498
8 Acetic acid Carboxylic acid 4.17 60
9 Acetic acid, 2,2-[oxybis(2,1-ethanediyloxy)]bis (2-[2-(Carboxymethoxy)ethoxy]ethoxy)acetic acid Carboxylic acid 4.24 222
10 Ethyl(dimethyl)isopropoxysilane Ethyl(dimethyl)silyl isopropyl ether Ether 4.54 146
11 Silane, triethylmethoxy- Methyl trethylsilyl ether Ether 4.6 146
12 Butanoic acid, 4,4-dithiobis[2-amino-,[S-(R*,R*)] Carboxylic acid 4.73 268
13 2-Pyridinecarboxylic acid Picolinic acid Carboxylic acid Natural chelator 4.73 123
14 2-Propanone, 1-(dimethylamino)- (Dimethylamino)acetone Ketone compound 4.89 101
15 2,2-Bioxirane Butane1,2:3,4-diepoxy- Heterocyclic compound 4.92 86
16 Cyclotrisiloxane, hexamethyl Dimethylsiloxane cyclic trimer Heterocyclic compound 5.3 222
17 Pyrimidine, 2-methyl- 2-Methylpyrimidine Heterocyclic compound 5.61 94
18 L-Galactose, 6-deoxy- 6-Deoxyhexose Sugar moiety Preservative 6.39 164
19 2-Propenamide Acrylamide Amide 6.43 71
20 1,2,4-Triazole, 4-(4-methoxybenzylidenamino)-5-methyl-3-(3,5-dimethylpyrazol-1-yl Heterocyclic compound 7.54 310
21 Acetic acid, 2-(dimethylamino)ethyl ester Dimethylaminoethanol acetate Ester 7.57 131
22 2H-1-Benzopyran, 3,5,6,8a-tetrahydro-2,5,5,8a-tetramethyl-,(2S-cis)- Edulan II Heterocyclic compound 7.58 192
23 Pyrazine, ethyl- Ethylpyrazine Heterocyclic compound 7.67 108
24 Oxirane, 2-ethyl-2-methyl Butane, 1,2-epoxy-2-methyl Heterocyclic compound 7.77 86
25 Butyrolactone Ketone compound 7.88 86
26 4-Methyl-2-haptanol Alcohol 7.96 130
27 1,2-Cyclopentanedione Ketone compound Antioxidant 7.98 98
28 Pyran-4-carboxylic acid, 4-(4-methoxyphenyl)-tetrahydro- Heterocyclic compound 8.02 236
29 6-Oxa-bicyclo[3.1.0]hexan-3-one Ketone compound 8.09 98
30 Dihydroxyacetone 2-Propanone, 1,3-dihydroxy- Ketone compound 8.18 90
31 Butanoic acid, 4-hydroxy- Carboxylic acid 8.54 104
32 Propanedioic acid, Propyl- 2-Propylmalonic acid Carboxylic acid 9.1 146
33 1,2,3-Propanetriol Glycerin Alcohol 9.33 92
34 2,4-Dihydroxy-2,5-dimethyl-3(2H)-furan-3-one Ketone compound 9.74 144
35 Oxirane, [(2-propenyloxy)methyl}- Propane, 1-(allyloxy)2,3-epoxy- Heterocyclic compound 10.26 114
37 HEPES[4-(2-Hydroxyethyl)-1-piperazineethanesuffonic acis)] Heterocyclic compound 10.26 238
37 2H-Pyran-2,6(3H)-dione Glutaconic anhydride Heterocyclic compound 10.75 112
38 1H-Indazole, 4,5, 6, 7-tetrahydro Heterocyclic compound 11.54 122
39 2-Pyrrolidinone, 1-methyl M-Pyrol Ketone compound 11.86 99
40 Benzeneacetaldehyde Aldehyde Antibacterial 12 120
41 2,4,6,-Cycloheptatrien-1-one,4-methyl- Ketone compound 12.06 120
42 2,5-Dimethyl-4-hydroxy-3(2H)-furanone Ketone compound 12.28 128
43 a-D-Glucopyranoside, O-a-D-glucopyranosyl-(1.fwdarw.3)-a-D-fructofuranosyl Sugar moiety 12.81 504
44 Phenol, 2-methoxy- Phenolic compound Antimicrobial, antioxidant, anti-inflammatory, analgesic 13.83 124
46 Formic acid, 3-methylbut-2-yl ester Ester 14.24 116
45 1-Butanol,3-methyl-, formate (isopentyl alcohol, formate) Isopentyl alcohol, formate Fatty acid ester Antimicrobial activity 14.24 116
47 1,5-Hexadien-3-ol Alkene 14.36 98
48 Cyclopentane, (1,1-dimethylethyl)-{Tert-Butylcyclopentane} Tert-Butylcyclopentane Alkane Antibacterial 14.68 126
49 4H-Pyran-4-one,3-hydroxy-2-methyl- Maltol Heterocyclic compound Flavor enhancer 14.78 126
50 5-Hepten-3-one, 5-methyl- Ketone compound 15.09 126
51 2-Acetyl-2,3,5,6-tetrahydro-1,4-thiazine 1-(3-Thiomorpholinylethanone Ketone compound 15.85 145
52 3,5-Dihydroxy-6-methyl-2,3-dihydro-4H-pyran-4-one Heterocyclic compound Antimicrobial, anti-inflammatory, anti-proliferative 16.61 144
53 Propanal, 2-(benzoyloxy)-,® 1-Methyl-2-oxoethyl benzoate Aldehyde 16.69 178
54 Benzoic Acid Carboxylic acid 16.76 122
55 N-aminomorpholine 4-Aminomorpholine Heterocyclic compound 16.95 102
56 Pentanoic acid, 2-isopropoxyphenyl ester 2-Isopropoxyphenyl pentanoate Ester 18.26 236
57 Phenol, 4-ethenyl-, acetate 4-Vinylphenyl acetate Phenolic compound Antimicrobial, antioxidant, anti-inflammatory, analgesic 19.29 162
58 Benzaldehyde, 3,4-dimethyl- 3,4-dimethylbenzaldehyde Aldehyde Antibacterial 19.3 134
59 Benzene, (ethenyloxy)- Ether, phenyl vinyl Ether 19.31 120
60 Benzofuran, 2,3-dihydro Coumaran Heterocyclic compound Antihelminthic, anti-inflammatory, anti-diarrhoeal 20.16 120
61 Benzeneacetaldehyde, 3-methyl m-Tolualdehyde Aldehyde Antimicrobial 20.34 120
62 1,2-Benzenediol,3-methoxy Pyrocatechol, 3-methoxy Phenolic compound Antioxidant 21.01 140
63 Isosorbide D-Glucitol, 1,4,3,6-dianhydro Alcohol 22.61 146
64 2-Methoxy-4-vinylphenol phenol, 4-ethenyl-2-methoxy- Phenolic compound Antimicrobial, antioxidant, anti-inflammatory, analgesic 23.35 150
65 (p-Hydroxyphenyl)glyoxal Benzeneacetaldehyde, 4-hydroxy-a-0x0 Aldehyde Antibacterial 23.71 150
66 2-Acetamido-2-deoxy-d-mannolactone Sugar moiety Anti-bacterial 24.8 217
67 Phenol, 2,6-dimethoxy- Pyrogallol 1,3-dimethyl ether Phenolic compound Antimicrobial, antioxidant, anti-inflammatory, analgesic 24.99 154
68 1-Undecanol alcohol Undecyl alcohol Alcohol 29.83 172
69 Phenol,2,6-bis(1,1-dimethylethyl)-4-methyl- Butylated hydroxytoluene Phenolic compound Antimicrobial, antioxidant, anti-inflammatory, analgesic 30.99 220
70 Phenol, 2,4-bis(1,1-dimethylethyl)- Phenol, 2,4-di-tert-butyl- Phenolic compound Antimicrobial, antioxidant, anti-inflammatory, analgesic 31.19 206
71 5-Tert-butyl-1,2,3-benzenetriol 5-Tert-butylpyrogallol Phenolic compound Antioxidant, antiseptic antibacterial, anti-dermatitic fungicide, pesticide 31.91 182
72 Benzoic acid, 4-ethoxy-, ethyl ester Ester 32.12 194
73 3,4-0-Isopropylidene-d-galactose 3,4,0-(1-Methylethylidene)hexopyranose Sugar moiety Preservative 32.35 220
74 Pentanoic acid, 2,2-4-trimethyl-3-carboxyisopropyl,isobutyl ester Ester 32.97 286
75 3,5-Dimethoxyacetophenone Ketone compound antioxidant 33.65 180
76 a-Methyl-D-mannopyranoside Sugar moiety Preservative 34.35 194
77 a-D-Galactopyranoside, methyl Galactopyranoside, methyl, a-D- Sugar moiety Preservative 34.61 194
78 3-O-methyl-d-glucose 3-O-methylhexose Sugar moiety Preservative 37.68 194
79 2,6-Diisopropylnapthalene Hetercyclic compound 37.71 212
80 Dodecyl acrylate n-Lauryl acrylate Ester 38.17 240
81 Cyclopenta [1,3][cyclopropa [1,2]cyclohepten-3(3ah)one, 1,2,3b,6,7,8-hexahydro-6,6-dimethyl- Ketone compound 40.31 190
82 5-Tert.butyloxy carboxamido-2,3,3-trimethyl-3H-indole Tert-butyl 2, 3,3-trimethyl-3H-indole-5-ylcarbamate Heterocyclic compound 41.6 274
83 Phthalic acid, hex-3-yl-isobutyl ester Ester 42.4 306
84 Hexadecanoic acid, methyl ester Palmitic acid, methyl ester Ester Antioxidant, flavor, hypocholesterolemic, nematicide 46.13 270
85 5,8, 11-Heptadecatriynoic acid methyl ester Ester 46.2 272
86 Phthalic acid, butyl undecyl ester Ester 47.36 376
87 1,2-Benzenedicarboxylic acid, dibutyl ester Dibutyl phthalate Ester Plasticizer, antimicrobial, antifouling 47.37 278
88 Methyl 10 trans, 12-cis-octadecadienoate Ester 48.53 294

Table S2.

List and basic features of identified phytocomponents in the methanolic extract of soybean genotypes by GCMS analysis.


  1. 1. FAOSTAT. Food and Agriculture Organization of the United Nations. 2016. Available from [Accessed: March, 2018]
  2. 2. Acquaah G. Breeding soybean. In: Acqaah G, editor. Principles of Plant Genetics and Breeding Book. Malden, MA, USA: Blackwell Publishing Ltd; 2007. p. 519
  3. 3. Lim TK. Edible Medicinal and Non-Medicinal Plants. Vol. 2, Fruits. New York: Springer Science, Business Media B.V; 2012
  4. 4. Gupta SK. Technological Innovations in Major World Oil Crops. New York, USA: Springer; 2012
  5. 5. Probst H, Judd RW. Origin, US history and development, and world distribution. In: Caldwell BE, editor. Soybean, Improvement, Production, and Uses. Madison, USA: American Society of Agronomy; 1973. pp. 1-15
  6. 6. Schmutz J, Cannon S, Schlueter J, Ma J, Mitros T, Nelson W, Hyten DL, Song Q, Thelen J, Cheng A, et al. Genome sequence of the paleopolyploid soybean. Nature. 2010;463:178-183
  7. 7. Pratap A, Gupta SK, Kumar J, Solanki RK, Soybean. In: Gupta SK, editor. Technological Innovations in Major World Oil Crops, Vol. 1: Breeding. New York: Springer Science Business Media; 2012. pp. 293-321. DOI: 10.1007/978-1-4614-0356-2_12
  8. 8. El-Shemy H, Soybean. Nutrition and health. In: El-Shemy, editor. Soybean-BioActive Compounds. Rijeka: InTech; 2013. pp. 453-473
  9. 9. Chung HY. Volatile components in fermented soybean ( Glycine max) curds. Journal of Agricultural and Food Chemistry. 1999;47(7):2690-2696
  10. 10. Chung HY, Fung PK, Kim JS. Aroma impact components in commercial plain sufu. Journal of Agricultural and Food Chemistry. 2005;53(5):1684-1691
  11. 11. Messina M. Soybean isoflavone exposure does not have feminizing effects on men, a critical examination of the clinical evidence. Fertility and Sterility. 2010;93:2095-2104
  12. 12. Luthria DL, Biswas R, Natarajan S. Comparison of extraction solvents and techniques used for the assay of isoflavones from soybean. Food Chemistry. 2007;105:325-333
  13. 13. Lee SJ, Kim JJ, Moon HI, Ahn JK, Chun SC, Jung WS, Lee OK, Chung IM. Analysis of isoflavones and phenolic compounds in Korean soybean Glycine max (L.) seeds of different seed weights. Journal of Agricultural and Food Chemistry. 2008;56:2751-2758
  14. 14. Gutierrez E, Wang T, Fehr WR. Quantification of sphingolipids in soybeans. Journal of the American Oil Chemists' Society. 2004;81:737-742
  15. 15. AOAC, editor. Official Methods of Analysis of the AOAC. 15th ed. Washington DC, USA: AOAC International; 1990
  16. 16. Markham R. Distillation apparatus suitable for micro-Kjeldahl analysis. The Biochemical Journal. 1942;36:970-791
  17. 17. Hammer O, Harper DA, Ryan PD. PAST, paleontological statistics software package for education and data analysis. Palaeontologia Electronica. 2001;4:1-9
  18. 18. Machado FP, Queróz JH, Oliveira MG, Piovesan ND, Peluzio MC, Costa NMB, et al. Effects of heating on protein quality of soybean flour devoid of Kunitz inhibitor and lectin. Food Chemistry. 2008;107:649-655
  19. 19. Zarkadas CG, Yu ZR, Voldeng HD, Minero-Amador A. Assessment of the protein quality of a new high-protein soybean cultivar by amino acid analysis. Journal of Agricultural and Food Chemistry. 1993;41:616-623
  20. 20. Zarkadas CG, Voldeng HD, Yu ZR, Choi V. Assessment of the protein quality of nine northern adapted yellow and brown seed coated soybean cultivars by amino acid analysis. Journal of Agricultural and Food Chemistry. 1999;47:5009-5018
  21. 21. Monteiro MR, Costa NM, Oliveira MG, Pires CV, Moreira MA. Qualidade proteica de linhagens de soja com ausência do inibidor de tripsina kunitz e das isoenzimas lipoxigenases. Revista de Nutrição. 2004;17:195-205
  22. 22. Liu K. Chemistry and nutritional value of soybean components. In: Liu K, editor. Soybeans, Chemistry, Technology and Utilization. New York, USA: Chapman & Hall; 1997. pp. 25-113
  23. 23. Messina M. Legumes and soybeans, overview of their nutritional profiles and health effects. The American Journal of Clinical Nutrition. 1999;70:439-450
  24. 24. Guillon F, Champ MM. Carbohydrate fractions of legumes, uses in human nutrition and potential for health. The British Journal of Nutrition. 2002;8:293-306
  25. 25. Karimi E, Oskoueian E, Hendra R, Jaafar HZ. Evaluation of Crocus sativus L. stigma phenolic and flavonoid compounds and its antioxidant activity. Molecules. 2010;15:6244-6256
  26. 26. Zheng W, Wang SY. Antioxidant activity and phenolic compounds in selected herbs. Journal of Agricultural and Food Chemistry. 2001;49:5165-5170
  27. 27. Skerget M, Kotnik P, Hadolin M, Hras AR, Simonic M, Knez Z. Phenols, proanthocyanidins, flavones and flavonols in some plant materials and their antioxidant activities. Food Chemistry. 2005;89:191-198
  28. 28. Duh PD, Tu YY, Yen GC. Antioxidative activity of water extracts of Harng Jyur (Chrysanthemum morifolium). Journal of Food Science and Technology. 1999;32:269-277
  29. 29. Aneta W, Jan O, Renate C. Antioxidant activity and phenolic compounds in 32 selected herbs. Food Chemistry. 2007;105:940-949
  30. 30. Tapiero H, Tew KD, Nguyen BG, Mathe G. Polyphenols, do they play a role in the prevention of human pathologies? Biomedicine & Pharmacotherapy. 2002;56:200-207
  31. 31. Wei Y, Liu ZQ, Liu ZL. Antioxidant effect of coumarin derivatives on free radical initiated and photosensitized peroxidation of linoleic acid in micelles. Journal of the Chemical Society, Perkin Transactions. 1999;2:969-974
  32. 32. Moleyar V, Narasimham P. Antifungal activity of some essential oil components. Food Microbiology. 1986;3:331-336
  33. 33. Kurita N, Miyaji M, Kurane R. Antifungal activity and molecular orbital energies of aldehyde compounds from oils of higher plants. Agricultural and Biological Chemistry. 1979;43:2365-2371
  34. 34. Kurita N, Miyaji M, Kurane R, Takahara Y. Antifungal activity of components of essential oils. Agricultural and Biological Chemistry. 1981;45:945-952
  35. 35. Yu AN, Sun BG, Tian DT, Qu WY. Analysis of volatile compounds in traditional smoke-cured bacon (CSCB) with different fibber coatings using SPME. Food Chemistry. 2008;110:233-238
  36. 36. Saegeman VS, Ectors NL, Lismont D, Verduykt B, Verhaegen J. Short and long term bacterial inhibiting effect of high concentration of glycerol used in preservation of skin allografts. Burns. 2008;34:205-211
  37. 37. Priya K, Vijaylakshmi VK. Determination of bioactive components of Cynodon dactylon by GC-MS analysis. New York Science Journal. 2011;4:16-20
  38. 38. Yang CS, Landau JM, Huang MT, Newmark HL. Inhibition of carcinogenesis by dietary polyphenolic compounds. Annual Review of Nutrition. 2001;21:381-406
  39. 39. Hedlund TE, Johannes WU, Miller GJ. Soy isoflavonoid equol modulates the growth of benign and malignant prostatic epithelial cells in vitro. The Prostate. 2003;54:68-78
  40. 40. Kamal M, Shakya AK, Jawid T. Benzofurans: A new profile of biological activities. International Journal of Medicine and Pharmaceutical Sciences. 2011;1:1-15
  41. 41. Shin E, Hwang CE, Lee BW, Kim HT, Ko JM, Baek IY, Lee Y, Choi J, Cho EJ, Seo WT, Cho KM. Chemometric approach to fatty acid profiles in soybean cultivars by principal component analysis (PCA). Preventive Nutrition and Food Science. 2012;17:184-191
  42. 42. Brodnjak-Voncina D, Kodba ZC, Novic M. Multivariate data analysis in classification of vegetable oils characterized by the content of fatty acids. Chemometrics and Intelligent Laboratory Systems. 2005;75:31-43
  43. 43. Zagonel GF, Peralta-Zamora P, Ramos LP. Multivariate monitoring of soybean oil ethanolysis by FTIR. Talanta. 2004;63:1021-1025
  44. 44. Brandao LF, Braga JW, Suarez PA. Determination of vegetable oils and fats adulterants in diesel oil by high performance liquid chromatography and multivariate methods. Journal of Chromatography. 2012;1225:150-157

Written By

Salem Alghamdi, Hussein Migdadi, Muhammad Khan, Ehab H. El- Harty, Megahed Ammar, Muhammad Farooq and Muhammad Afzal

Submitted: 25 March 2018 Reviewed: 27 April 2018 Published: 07 November 2018