Validasi Metode dan Penetapan Kadar Genistein Ekstrak

Edamame (Glycine Max L. Merrill) Terfermentasi Rhizopus

oligosprorus dan Kombinasi dengan Aspergillus oryzae
menggunakan KLT Densitometri
(Validation Methods and Determination of Genistein Content of
Edamame (Glycine Max L. Merrill) Extract Fermented by Rhizopus
oligosprorus and Combination with Aspergillus oryzae by TLC
Densitometry)
Endah Puspitasari, Ayu Maulida Fitriah, Dewi Novitasari
Fakultas Farmasi, Universitas Jember
Jln. Kalimantan 37, Jember 68121
e-mail korespondensi: e.puspitasari@gmail.co.id

Abstract
on of genistein content in edamame extract fermented for the first, second, third and fourth days respectively
were 0,0047 ± 1,42 x10-4; 0,0062 ± 2,19 x10-4; 0,0106 ± 2,38 x10-4; and 0,0183 ± 2,08 x10-4% w/w.
Edamame (Glycine max L. Merrill) is a type of Japanese soybeans that contain isoflavone aglikon such
as genistein with activity as anticancer. The genistein content can be improved by fermentation. The aim of this
research was to determine the content of genistein in edamame fermented by R. oligosporus and combination
with A. oryzae using TLC-densitometry method that had been validated previously. Analytical conditions used
include methanol p.a as solvent; n-hexane: ethyl acetate: acetic acid (2: 5: 0,15) as eluent ; Silica Gel 60 F 254 as
stationary phase; wavelength 266 nm; and test concentration 5-50 μg/ml. Validation parameters include linearity;
LOD and LOQ; specificity; precision and accuracy is fulfill the requirements. The selectivity parameter was not
fulfill (Rs <1,5). The results of determination content on edamame non-fermentation and fermentation by R.
oligosporus on day 1; 2; 3; and 4 ± SD respectively are 0,01887 ± 2,6x10-4; 0,00301 ± 4,1x10-4; 0,01545 ±
5,9x10-4; 0,00772 ± 3x10-4; and 0,00109 ± 3,9x10-4% (w/w), while fermented by combination with . (p <0.01).

Keywords: genistein, edamame, Rhizopus oligosporus, Aspergillus oryzae, TLC-densitometry

Abstrak
Isoflavon merupakan senyawa fitoestrogen bersifat antikanker, yang banyak terkandung dalam kedelai,
seperti edamame (Glycine max L. Merril). Isoflavon dalam bentuk aglikon, salah satunya genistein, mempunyai
aktivitas biologis lebih tinggi daripada dalam bentuk glikosida. Untuk meningkatkan kadar genistein tersebut
dilakukan fermentasi dengan kombinasi Aspergillus oryzae dan Rhizopus oligosporus. Tujuan dari penelitian ini
adalah untuk menetapkan kadar genistein dalam edamame yang difermentasi, dengan metode KLT densitometri
yang sebelumnya divalidasi terlebih dahulu dengan parameter linieritas, LOD dan LOQ, selektivitas/spesifisitas,
presisi, dan akurasi. Sebelum validasi, dicari kondisi analisis yang optimal, yaitu metanol p.a sebagai pelarut,
fase diam berupa silika gel 60 F 254, eluen berupa n-heksana: etil asetat: asam asetat (2:5:0,15), panjang
gelombang optimal 266 nm, dan konsentasi uji 5,6; 6; 15,6; 17,8; 42 µg/ml µg/ml. Semua parameter validasi
memenuhi syarat, kecuali selektivitas. Hasil penetapan kadar genistein dalam ekstrak edamame terfermentasi
hari pertama, kedua, ketiga dan keempat secara berurutan adalah 0,0047 ± 1,42 x10-4; 0,0062 ± 2,19 x10-4;
0,0106 ± 2,38 x10-4; dan 0,0183 ± 2,08 x10-4 %b/b.

Keywords: genistein, edamame, Rhizopus oligosporus, Aspergillus oryzae, KLT-densitometri

Akan dipublikasikan di Jurnal Internasional Der Pharma Chemica

Edamame (Glycine max L. Merrill) is a type of Japanese soybeans that contain isoflavone aglikon such as
genistein with activity as anticancer. The genistein content can be improved by fermentation. The aim of this
research was to determine the content of genistein in edamame fermented by R. oligosporus and combination
with A. oryzae using TLC-densitometry method that had been validated previously. Analytical conditions used
include methanol p.a as solvent; n-hexane: ethyl acetate: acetic acid (2: 5: 0,15) as eluent ; Silica Gel 60 F 254 as
stationary phase; wavelength 266 nm; and test concentration 5-50 μg/ml. Validation parameters include linearity;
LOD and LOQ; specificity; precision and accuracy is fulfill the requirements. The selectivity parameter was not
fulfill (Rs <1,5).