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Fenugreek Seeds Essay

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3. 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 40±5 °C in a hot air oven to increase its keeping quality and stored in air tight containers at ambient temp.

Raw fenugreek seeds

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 until the constant weight. The average length of sprouts was 1.1cm,1.7cm and 2.6cm which was germinated
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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 in orbital shaker for 4 hrs at 45°C. In extraction process 3 g of prepared sample were weigh 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 phenolic content 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 Folin-Ciocalteu reagent (diluted 2 times with water) was added and, then, 1 ml of Na2CO3 (40 %) 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 blue-colored 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 mg/g of
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One g of material was ground and extracted with 0.2 N HCl by continuous shaking. To 0.2 ml of the filtrate, distilled water to make volume 1.4 ml was added. After that1ml of ferric ammonium sulphate solution (21.6 mg in 100 ml water) was added, mixed and placed 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/100 g of
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