IJSTE - International Journal of Science Technology & Engineering | Volume 3 | Issue 06 | December 2016

ISSN (online): 2349-784X

Effect of Germination and Roasting on Nutritive

Composition and Anti-Nutrients in Fenugreek
Seeds
Manju
GJU Hisar Haryana

Dr. B. S. Khatkhar
GJU Hisar Haryana
Mrs. Parveen
GJU Hisar Haryana

Abstract
Fenugreek seeds were processed by germination and roasting. Raw and processed fenugreek seeds were analyzed for total
polyphenolic content, antioxidant activity and tannins in 80% methanolic extract and absolute methanolic extract. Total
polyphenolic content, antioxidant activity and tannins were significantly (P 0.05) different in 80% methanolic extract and
absolute methanolic extract. Germination process significantly (P 0.05) increased total polyphenolic content and antioxidant
activity. The maximum level of phenolic content (93.27 mg/g) was found in fenugreek seeds germinated for 48 hrs. Total
polyphenolic content, antioxidant activity and tannins were also significantly (P 0.05) different in roasted fenugreek seeds.
Keywords: Fenugreek seeds, Germination process, Estimation of phytic acid
________________________________________________________________________________________________________
I.

INTRODUCTION

Herb and spices have been used in food as a food additives due to their natural antioxidant property. Fenugreek (Trigonella
foenum-gracum) is an annual leguminous bean member of Fabaceae family. Its seeds and green leaves are having many
medicinal applications (Thomas et al., 2011; Paridar et al., 2011; Vaidya etal., 2013). In India, it is extensively used as
Ayurvedic medicine and in China as traditional medicine (Prasad et al., 2014). Interestingly, in herbal medicine, it is used in the
treatment of diabetes (Leela and Shafeekh, 2008).Fenugreek leaves provide a good amount of various minerals and vitamins.
They are rich in choline.
Medicinally seeds are the most important and useful part of fenugreek plant .Seeds are aromatic, bitter, carminative,
galactogogue and have antibacterial properties.These seeds are golden-yellow in colour, small in size, hard, having four-faced
stone like structure. Fenugreek seeds have high protein (25 %), lysine (5.7 g/16 g N), soluble (20 %) and insoluble (28 %) dietary
fiber besides being rich in calcium, iron and betacarotene. The biological and pharmacological actions of fenugreek seeds are
mostly attributed to the variety of its bioactive chemical constituents that serve as raw materials for the manufacturing of various
hormonal and therapeutic drugs (Mehrafarin etal., 2010; Priya et al., 2011). Their application are limited due to bitter taste
(Sharma 1986). However these days various processing techniques are applied for debittering of fenugreek seeds.
Secondary metabolites flavonoids and phenolic compounds are widely distributed in plant and it exert multiple biological
effects including free radical scavenging, anti-inflammatory, and anticarcinogenic (Miller A. L.,1996).T. foenum-graecum
contain high phenolic contents having high antioxidant activity(Kaur.C.et al.,2002).Among the plethora of bioactive compounds
found in fenugreek seeds the major chemical constituents are polyphenolic compounds, galactomannan (fiber), diosgenin
(saponin) , quercetin (flavonoid), trigonelline (alkaloid) and 4-hydroxyisoleucine (unusual amino acid). Germinated fenugreek
seed are rich in polyphenolic compounds. Being a rich source of mucilaginous fiber and other dietary essential, fenugreek seeds
used as a functional and nutritional food.
Therefore, the present study was planned to evaluate raw, germinated and roasted fenugreek seed extracts as a source of
natural antioxidant. In this study, the effect of germination and roasting on their chemical composition was also investigated.
II. MATERIALS AND METHODS
Fenugreek seeds were purchased from local market of Hissar, Haryana. Seeds were cleaned to remove any extraneous material.
Raw seeds were dried at 405 C in a hot air oven to increase its keeping quality and stored in an airtight containers at ambient
temp.

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Effect of Germination and Roasting on Nutritive Composition and Anti-Nutrients in Fenugreek Seeds
(IJSTE/ Volume 3 / Issue 06 / 007)

Germination
Fenugreek seeds (20 g) were soaked overnight in water at the ratio of 1:5 (w/v). The excess water was drained and seeds were
germinated (tied in a muslin cloth) at room temperature for 24 h, 36 h and 48 h. The germinated seeds were dried in an oven at
50 C til the constant weight.
Roasting
Fenugreek seeds (20 g) were roasted in an open pan at 1305 C ,1505 C and 2005Cfor 7 min,5 min and 2 min respectively.
Continuous stirring was done with laddle for proper and uniform roasting.
Preparation of Fenugreek Seeds Extract
Raw and processed (germination and roasting) fenugreek seeds were ground in Wall Mill. Ground sample were collected in air
tight container separately for further analysis at room temperature. The prepared sample were extracted in absolute methanol and
80% methanol using orbital shaker for 4 hrs at 45C. In extraction process, 3 g of prepared sample were weighed in universal
bottle and 75 ml solvent was added .After extraction process supernatant were collected for further analysis.
Determination of Total Phenolic Content
The total phenols in extracts was measured by UV spectrophotometry based on a colorimetric oxidation/reduction reaction. The
oxidizing agent used was Folin-Ciocalteu reagent (AOCS,1990). To 0.2 ml of extract,7.5 ml water was added in a test tube and
after that 0.5 ml of 1 N Folin-Ciocalteu reagent was added and, then, 1 ml of Na 2CO3 were added. The sample was incubated
for 30 min at room temperature . For a control sample, 0.2 ml of distilled water was used. The absorbance of the resulting bluecolored solutions was measured at 760 nm.Quantitative measurements were performed, based on a standard calibration curve of
gallic acid in water. The results were expressed as gallic acid equivalents (GAE) in milligrams per gram of sample.
Determination of Total Flavonoid Content
Total flavonoid content was determined using a method by (Liu et al., 2008).Briefly, 2 ml of the extracts was taken in a test tube
and 0.2 ml of 5% sodium nitrite was mixed.After 5 min 0.2 ml of 10%aluminium chloride was added and after 6 min 1 ml of
1M sodium hydroxide was added. The absorbance was measured at 510 nm.The resultswere expressed as Quarcetin equivalents
(QE) in milligrams per gram of sample.
Total Dietary Fiber
The dietary fiber content in processed fenugreek seeds was estimated by rapid enzymatic assay method. Briefly, 1 g processed
fenugreek seeds sample was subjected to sequential enzymatic digestion by heat-stable -amylase, protease and
amyloglucosidase. Insoluble dietary fiber (IDF) filtered, and then residue were washed with warm distilled water. Combined
solution of filtrate and water washings was precipitated with 4 volumes of 95% ethanol (EtOH) for soluble dietary fiber (SDF)
determination. Precipitates was filtered and dried. Both SDF and IDF residues were estimated for protein, ash and blank, for the
final calculation of SDF and IDF values. SDF was precipitated with EtOH,and residue was filtered, dried and weighed. Total
dietary fiber (TDF) value was calculated for protein and ash content (AOAC, 1991).
Total Antioxident Activity
Antioxidant activity was estimated by DPPH free radical scavenging method (Maizura et al.,2011). To 0.2 ml of extract taken, 2
ml DPPH in methanol was added and volume was made 3 ml. After that sample was incubated for 30 min in dark. Absorbance
was measured at 517nm.
Total antioxidant activity was expressed as the % of DPPH decrease using the equation:
% Antioxidant activity= (Control absorbancesample absorbance) 100
Control absorbance
Estimation of phytic acid
Phytic acid was estimated by the method of Davies and Reid (1979). One g material was ground and extracted with 0.2 N HCl.
To 0.2 ml of the extract, distilled water to make volume 1.4 ml was added. After that 1 ml of ferric ammonium sulphate solution
was added, mixed and kept in a boiling water bath for 20 min. The contents were cooled and 5 ml of isoamyl alcohol was added
and mixed. To this, 0.1 ml ammonia solution was added, shaken thoroughly and centrifuged at 3000 rpm for 10 min. The
alcoholic layer was separated and the colour intensity was read at 465 nm against amyl alcohol blank after 15 min. Sodium
phytate standards were run along with the sample. The results were expressed as mg phytic acid/100 g sample.

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Effect of Germination and Roasting on Nutritive Composition and Anti-Nutrients in Fenugreek Seeds
(IJSTE/ Volume 3 / Issue 06 / 007)

Estimation of tannins
Tannins were estimated by Vanillin-HCl method (Price et al.,1978). One ml of suitably diluted extract was taken in a test tube
and 5 ml of freshly prepared vanillin-HCl reagent was added slowly with mixing and colour developed was read at 525 nm.
Catechin standards were run simultaneously along with sample. The results were expressed as mg/100g.
Statistical analysis
Each experiment was conducted in triplicates. The results were expressed as meanSD. Analysis of variance was applied to
analyze data and significance was accepted at p0.05 level.
III. RESULT AND DISCUSSION
Total Phenolic Content
Data pertaining to total phenolic contents presented in Table 1.1.Total phenolic content of raw and processed fenugreek seeds
was significantly (P 0.05) different in absolute methanolic extract and 80% methanolic extract. This difference might be due
difference in polarity of extracting solvent (Rostagno.M.A.et.al., 2003). Total phenolic content of raw fenugreek seeds was 67.32
mg/g and 59.72 mg/g in 80% methanolic extract and aqueous methanolic extract respectively. Total phenolic contents
significantly (P 0.05) increase by germination process. In 80% methanolic extract phenolics content increased from 67.32 mg/g
to 75.17 mg/g, 80.25 mg/g and 93.27 mg/g in samples germinated for 24 hr, 36hr and 48 hr respectively. In absolute methanolic
extract phenolic content increased from 59.72 mg/g to 89.54 mg/g in germinated fenugreek seeds.The highest level of phenolic
content (93.27 mg/g) was observed in fenugreek seeds germinated for 48 hr in 80% methanolic extract. Increases might be due to
the biosynthesis of phenolic compounds during germination process (Randhir et al., 2004).Total phenolic content in roasted
fenugreek seeds increased significantly (P 0.05) from 73.12mg/g to 86.43mg/g in 80% methanolic extract as presented in
table 1.1. In roasting process TPC increased with roasting temp. and was oberved maximum (86.43 mg/g) at 200C temp (Jeong
et al.,2004).

Solvent
80%Methanol
Methanol

Raw
67.320.22b
59.720.01a

Table 1
Effect of processing on TPC in processed fenugreek seeds.
TPC(mg/g)
Germination
Roasting
24 hr
36 hr
48 hr
130C
150C
75.170.06b 80.250.13b 93.270.33b 73.120.06b 81.300.44b
66.640.14a 72.600.10a 89.540.05a 61.340.36a 64.120.09a

200C
86.430.16b
76.640.22a

Means in the same column with different superscripts differ significantly at (P < 0.05).
Antioxident activity
Antioxidant activities of raw and germinated fenugreek seeds as shown in Table 1.2. DPPH radical-scavenging activity
expressed in % inhibition of raw and germinated fenugreek seeds ranged from 39.11% to 73.65% in 80% methanolic extract.
Antioxident activity is significantly (P 0.05) different in both extracting solvent of raw and processed fenugreek seeds. In
absoulte methanolic extract antioxidant activity ranged from 16.47% to 63.72%. Germinated fenugreek seeds had significantly (P
0.05) higher DPPH radical-scavenging activity compared to raw seeds. Polyphenols have high free radical scavenging
activity.This increase might be dueto the synthesis of compounds and tocopherols which are responsible for antioxidant activity
(Sharma and Gujral, 2010).
Antioxident activity increased from 39.11% to 49.03%,54.19% and 61.29% in germinated and roasted at 130C,150C and
200C respectively in 80% methanolic extract. In absoult methanolic extract, antioxidant activity ranged from 29.11% to
44.31%.

Solvent
80%Methanol
Methanol

Table 2
Effect of processing on AOA in processed fenugreek seeds.
Antioxident Activity(%)
Germination
Roasting
Raw
24 hr
36 hr
48 hr
130C
150C
39.110.08a 50.740.14a 53.870.12a
73.650.29a 49.03 0.14a 54.19 0.09a
16.470.19b 35.370.13b 53.410.21b
63.720.31b
29.11 0.11b 41.060.07b

200C
61.29 0.04a
44.310.23b

Means in the same column with different superscripts differ significantly at (P < 0.05).
Total Flavonoid Content
Data pertaining total flavonoids content is presented in Table 2. Total flavonoids content of raw fenugreek seeds was 3.48 mg/ g
in 80% methanolic extract. Total flavonoids content in 80% methanolic extract was increased to 5.29 mg/g and in absoulte
methanolic extract it increased to 5.10 mg/g in processed fenugreek seeds.

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33

Effect of Germination and Roasting on Nutritive Composition and Anti-Nutrients in Fenugreek Seeds
(IJSTE/ Volume 3 / Issue 06 / 007)

Solvent
80%Methanol
Methanol

Table 3
Effect of processing on total flavonoids content of processed fenugreek seeds.
TFC(mg/g)
Germination
Roasting
Raw
24 hr
36 hr
48 hr
130C
150C
3.480.08b 4.980.04b 5.17 0.06b 5.290.10b 3.610.08b 3.72 0.23b
3.150.09a 4.20 0.14a 4.40 0.09a 5.100.04a 3.190.17a 3.38 0.02a

200C
3.92 0.01b
3.540.19a

Means in the same column with different superscripts differ significantly at (P < 0.05).
Total flavonoids content of raw and processed fenugreek seeds significantly (P 0.05) different in two different extracting
solvent. Flavonoids content in raw fenugreek seeds was significantly (P 0.05) lower than germinated seeds. Significantly (P
0.05) increase in the total flavonoids content of fenugreek seeds was observed as a result of germination process. A gradual
increase was observed in flavanoid content with increase in germination time from3.48 mg/g to 5.29 mg/g . The highest level
(5.29 mg/g ) was recorded for fenugreek seeds germinated for 48 hr. Roasting process also increased the flavonoids content from
3.48 mg/g to 3.92 mg/g in 80%methanolicextract.In absoulte methanolic extract, flavonoids content also increased from 3.15
mg/g to 3.54 mg/g.
Antinutritional Factors
Significant (P 0.05) difference was observed in tannin content extractced in two solvents in as shown in Table 3. Tannins
content was higher in absolute methanolic extract (41.76mg/100g) and reduced to 27.23 mg/100g during germination process. In
germination process, tannins content was decreased from 12.52 mg/100g to 9.40mg/100g in fenugreek seed in 80% methanolic
extract. As the germination period increased tannin content gradually decreased. The reduction might be due to leaching of
tannins into water (Shimelis and Rakshit, 2007) and bonding of polyphenols with carbohydrate or protein (Saharan et al., 2002).
In roasting process, tannins were decreased from 12.52mg/100g to 8.42mg/100g in 80% methanolic extract and from 41.76
mg/100g to 20.37 mg/100g in absolute metabolic extract. During roasting breakdown of the bond between phytate and might
takes place which results in destruction of phytates, tannins and oxalates (Reddy et al., 1978)

Solvent
80%Methanol
Methanol

Table 4
Effect of processing on tannins content in processed fenugreek seeds.
Tannins(mg/100g)
Germination
Roasting
Raw
24 hr
36 hr
48 hr
130C
150C
12.520.21a 11.220.22a 10.310.38a
9.400.57a 9.220.23a
8.450.49a
41.760.30b 33.370.28b 31.750.34b 27.230.15b 38.610.07b 32.17 0.14b

200C
8.42 0.15a
20.370.24b

Means in the same column with different superscripts differ significantly at (P < 0.05).
Germination of fenugreek seeds reduced the phytic acid content from 190.30-117.74 mg/100 g. It was reported that phytase
activity originates after germination and the phosphatase activity was increased in the germinated seeds which results in the
reduction of phytic acid content in fenugreek seeds after germination and roasting ( El Mahdy and El-Sebaiy .,1982).During
germination enzymatic hydrolysis of phytate phosphorus takes place which decreases phytic acid content (Gupta et al.,
2001).During roasting phytic acid reduced from298.77-177.18 mg/100g.Decrease in phytic acid during roasting may be due to
thermoability of phytic acid.Germination effects phytic acid more than roasting as shown in Table.4
Total Dietary Fibre
Data regarding TDF, IDF and SDF presented in Table 4. Significant(P 0.05) reduction was noted in total dietary fibre
(TDF),soluble dietary fibre (SDF) and insoluble dietary fibre (IDF) upon germination as shown in Table 4.Total dietary fibre
reduced from 47.35% to 40.20% during germination process.
Table 5
Effect of processing on Dietary Fibre and phytic acid in processed fenugreek seeds.
Sample
Raw
Germination
24 hr
36 hr
48 hr
Roasting
130C
150C
200C

TDF (%)
50.200.14a

IDF (%)
31.700.14a

SDF (%)
18.400.14b

PA (mg/100g)
317.180.14a

47.350.21b
43.400.28d
40.200.14f

30.300.14b
28.200.14c
28.450.35c

17.600.14c
15.650.21d
11.500.28e

190.300.25c
157.440.14f
117.740.24g

45.450.2c
41.400.28e
32.600.28g

24.450.35c
23.450.35e
20.650.21f

20.750.21a
17.450.35c
11.550.35e

298.770.31b
182.250.07d
177.180.21e

Means in the same column with different superscripts differ significantly at (P < 0.05).
There was no significant (P 0.05) change in insoluble dietary fibre germinated for 36 hr and 48 hr. Soluble dietary fibre also
significantly (P 0.05) reduced from 17.60% to 11.50% upon germination process. Reduction in dietary fiber content during
germination may be due to enzymatic breakdown of the galactomannan units. Shakuntala et al.(2011) concluded that decrease in
soluble dietary fiber content on germination of fenugreek seeds. An enzyme -galactosidase during germination of fenugreek
seeds partially attacks galactomannan to produce galactose. Decrement in total dietary fiber, insoluble dietary fiber and soluble

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34

Effect of Germination and Roasting on Nutritive Composition and Anti-Nutrients in Fenugreek Seeds
(IJSTE/ Volume 3 / Issue 06 / 007)

dietary fiber also noticed during roasting process of fenugreek seed as shown in Table 4. Total dietary fibre (TDF) reduced from
45.45% to 32.60% during roasting process. Significant (P 0.05) reduction in soluble dietary fibre (SDF) and insoluble dietary
fibre (IDF) also determined as listed in table 4.Reduction in IDF content during roasting might bedue to retrogradation of starch
(Mathur and Chaudhary.,2009).
IV. CONCLUSION
It may be concluded from the present study that nutritional and functional quality of fenugreek seeds can be improved by various
processing methods e.g germination and roasting. Antioxidant activity also increases significantly after processing which are
found to be responsible for functional properties of processed fenugreek seeds. Therefore, the use of processed fenugreek seeds
can be exploited in functional foods as well as a therapeutic agent.
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