RESTU KUMALASARI, 050710192 (2011) VALIDASI METODE KCKT UNTUK PENETAPAN KADAR GENISTEIN DALAM BISKUIT KEDELAI (Glycine max (L) Merr.). Skripsi thesis, UNIVERSITAS AIRLANGGA.

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Abstract

Genistein is the most isoflavone commonly found in soybeans and has the greatest estrogenic activity amongst other isoflavones. The aim of the present work was to validate the assay method of genistein in Soy biscuits by HPLC and to determine the concentrations of genistein in Soy biscuits with HPLC methods. In this research, method development for assay of genistein in Soy biscuits by using HPLC will be studied. The extract were examined by HPLC using LiChrospher RP-18 column, 4 mm x 250 mm x 5 μm and the mobile phase of acetonitrile and 10% acetic acid with gradient system, acetonitrile 20% within 0 – 15 minutes, 70% within 15 - 25 minutes, 20% within 25-30 minutes with a flow rate of 0.8 mL/minute and UV detection was perfor med at 260 nm. The resolution factor (Rs) between genistein and other com pound was 4.44 and the selectivity factor ( α ) was 1.07. Good linier relationship (r = 0.μ9999) between peak area and concentrations was found at range of genistein 1.98 ppm – 19.83 ppm . Genistein recoveries were 91.55% - 105.37% with coefficient of variation (CV) 4.49%. These result demonstrate that genistein in Soy biscuits can be detected by HPLC method. By using this method, genistein conc entrations in Soy biscuits as (6.30 ± 0.43) x 10 -4 % or 188.95 ± 12.90 μg/30 gram

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