ISOLASI DAN ANALISIS GENISTEIN DARI TEMPE BUSUK MENGGUNAKAN METODE KROMATOGRAFI KOLOM

Hartati Soetjipto, Yohanes Martono, Zulfa Yuniarti

Abstract


Isolation and Analysis of Genistein of Overripe Tempe using Column Chromatography Method

ABSTRACT

Genistein is one of the aglycone isoflavone compounds in tempe that has various biochemical activities, including anticancer, antitumor, and antioxidants. Commonly used isoflavone extraction methods resulted in isoflavone crude extract. The aim of this study was to isolate the genistein of overripe tempe through determining the appropriate combination of mobile phases in genistein isolation and the determination of genistein content in both crude extract and isolate. The overripe tempe was first extracted, then genistein was isolated from the crude extract using column chromatography method. The determination of mobile phase combination was done by Thin Layer Chromatography while the genistein content was quantitatively determined by using High Performance Liquid Chromatography. The results showed that the appropriate combination of mobile phase for genistein isolation was chloroform : methanol (15 : 1, v/v). Genistein content in the crude extract and isolates were 4737.50 and 31.36 μg/g extract, respectively. The genistein purity in the isolates was 63.80%, while the purity in the isoflavone extract was 31.98%.

Keywords: genistein, HPLC, isoflavone, overripe tempe, TLC

 

ABSTRAK

Genistein merupakan salah satu senyawa isoflavon aglikon dalam tempe yang memiliki bermacam-macam aktivitas biokimia, diantaranya antikanker, antitumor, dan antioksidan. Metode ekstraksi isoflavon yang umum diterapkan, menghasilkan ekstrak kasar isoflavon yang masih berupa campuran. Tujuan dari penelitian ini adalah untuk mengisolasi genistein dari tempe busuk melalui tahap penentuan kombinasi fase gerak yang tepat dalam isolasi genistein serta penentuan kandungan genistein baik dalam ekstrak kasar maupun isolat. Tempe busuk mula-mula diekstrak, selanjutnya genistein diisolasi dari ekstrak kasar menggunakan metode kromatografi kolom. Penentuan kombinasi fase gerak dilakukan secara Kromatografi Lapis Tipis, sedangkan kandungan genistein secara kuantitatif ditentukan dengan menggunakan Kromatografi Cair Kinerja Tinggi. Hasil penelitian menunjukkan bahwa kombinasi fase gerak yang tepat untuk isolasi genistein adalah kloroform : metanol (15 : 1, v/v). Kandungan genistein dalam ekstrak kasar dan isolat genistein berturut-turut sebesar 4737,50 dan 31,36 μg/g ekstrak. Kemurnian genistein dalam isolat adalah sebesar 63,80%, sedangkan kemurniannya dalam ekstrak isoflavon adalah sebesar 31,98%. 

Kata Kunci: genistein, HPLC, isoflavon, tempe busuk, KLT

Keywords


genistein; HPLC; isoflavone; overripe tempe; TLC

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References


Ahmad A, Ramasamy K, Majeed ABA, Mani V (2015) Enhancement of β-secretase inhibition and antioxidant activities of tempeh, a fermented soybean cake through enrichment of bioactive aglycones. Pharm Biol 53:758–766. doi: 10.3109/13880209.2014.942791

Ariani SRD, Hastuti W (2009) Analisis isoflavon dan uji aktivitas antioksidan pada tempe dengan variasi lama waktu fermentasi dan metode ekstraksi. Prosiding Seminar Nasional Kimia dan Pendidikan Kimia, Pp 568–580. 18 Maret 2009, Surakarta

Asih IARA (2009) Isolasi dan identifikasi senyawa isoflavon dari kacang kedelai (Glycine max). J Kimia 3:33–40

Astuti S (2008) Isoflavon kedelai dan potensinya sebagai penangkap radikal bebas. J Tek Industri dan Hasil Pertanian 13:126–136

Bele AA, Khale A (2011) An overview on thin layer chromatography. Int J Pharma Sci Res 2:256-267. doi: 10.13040/IJPSR.0975-8232.2(2).256-67

Beller NR, Hilleary CJ (1976) A sample preparation technique for column chromatography. J Chem Educ 53: 498. doi: 10.1021/ed053p498

Bhagwat S, Haytowitz DB, Holden JM (2008) USDA database for the isoflavone content of selected foods: Release 2.0. Nutrient Data Laboratory, United State of America

Cheng KC, Lin JT, Wu JY, Liu WH (2010) Isoflavone conversion of black soybean by immobilized Rhizopus spp. Food Biotechnol 24: 312-331. doi: 10.1080/08905436.2010.524459

César IC, Braga FC, Soares CD, Nunan EA, Pianetti GA, Condessa FA, Barbosa TA, Campos LM  (2006) Development and validation of a RP-HPLC method for quantification of isoflavone aglycones in hydrolyzed soy dry extracts. J Chromatogr B 836:74–78. doi: 10.1016/j.jchromb.2006.03.030

Hessler PE, Larsen PE, Constantinou AI, Schram KH, Weber JM (1997) Isolation of isoflavones from soy-based fermentations of the erythromycin-producing bacterium Saccharopolyspora erythraea. Appl Microbiol Biotechnol 47:398–404. doi: 10.1007/s002530050947

Hong GE, Mandal PK, Lim KW, Lee CH (2012) Fermentation increases isoflavone aglycone contents in black soybean pulp. Asian J Animal Vet Adv 7:502-511. doi: 10.3923/ajava.2012.502.511

Jyoti, Agrawal SS, Saxena S, Sharma A (2015) Phytoestrogen "genistein": Its extraction and isolation from soybean seeds. Int J Phramacogn Phytochem Res 7:1121-1126

Kuligowski M, Pawłowska K, Jasińska-Kuligowska I, Nowak J (2016) Isoflavone composition, polyphenols content and antioxidative activity of soybean seeds during tempeh fermentation. CyTA J Food 15:27-33. doi: 10.1080/19476337.2016.1197316

Lee IH, Chou CC (2006) Distribution profiles of isoflavone isomers in black bean kojis prepared with various filamentous fungi. J Agric Food Chem 54:1309-1314. doi: 10.1021/jf058139m

Lewidharti RS, Soetjipto H, Andini S (2015) Dinamika konsentrasi genistein dalam proses pembusukan tempe kedelai. Prosiding Seminar Nasional Kimia dan Pendidikan Kimia VII. 18 April 2015, Surakarta

Purnama FA, Dewi L, Hastuti SP (2012) Kadar air, abu, protein dan karbohidrat pada tahap pembuatan tempe. Skripsi, Universitas Kristen Satya Wacana

Riyanto CA, Soetjipto H (2017) Solvent optimization for genistein isolation of “rotten tempe” by high performance liquid chromatography method. J Eksakta: 17:111-118

Rosida DF, Sudaryati HP, Constantia F (2009) Kajian peran angkak pada kualitas tempe kedelai-lamtoro gung (Leucaena leucocephala). J Rekapangan 6:64-72

Rostagno MA, Villares A, Guillamon E, Garcia-Lafuente A, Martinez JA (2009) Sample preparation for the analysis of isoflavones from soybeans and soy foods. J Chromatogr A 1216:2–29. doi: 10.1016/j.chroma.2008.11.035

Sartini, Djide MN, Permana AD, Ismail (2014) Ekstraksi isoflavon kedelai dan penentuan kadarnya secara Ultra Fast Liquid Chromatografi (UFLC). J Sainsmat 3:130-134. doi: 10.2685/sainsmat3211202014

Sherma J, Fried B (2003) Handbook of Thin-Layer Chromatography, Third Edition. Marcel Dekker Inc, New York

Shewiyo DH, Kaale E, Risha PG, Dejaegher B, Smeyers-Verbeke J, Vander-Heyden Y (2012) Optimization of a reversed-phase-high-performance thin-layer chromatography method for the separation of isoniazid, ethambutol, rifampicin and pyrazinamide in fixed-dose combination antituberculosis tablets. J Chromatogr A 1260:232–238. doi: 10.1016/j.chroma.2012.08.044

Singh-Gupta V, Zhang H, Yunker CK, Ahmad Z, Zwier D, Sarkar FH, Hillman GG (2010) Daidzen effect on hormone refractory prostate cancer in vitro and in vivo compared to genistein and soy extract: Potentiation of radiotherapy. Pharm Res 27:1115-1127. doi: 10.1007/s11095-010-0107-9

Suharto KF, Soetjipto H, Martono Y (2017) Pengaruh lama fermentasi tempe terhadap kandungan total senyawa fenolik dan isoflavon genistein. ALCHEMY J Penelitian Kimia 13:230-240. doi: 10.20961/alchemy.v13i2.5094

Yunindarwati E, Ulfa EU, Puspitasari E, Hidayat MA (2016) Penentuan kadar genistein dan aktivitas hambatan tirosinase kedelai (Glycine max) terfermentasi Aspergillus oryzae. J Ilmu Kefarmasian Indones 14:1–7




DOI: http://dx.doi.org/10.29122/jbbi.v5i1.2860

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