ISOLASI DAN ANALISIS GENISTEIN DARI TEMPE BUSUK MENGGUNAKAN METODE KROMATOGRAFI KOLOM
Main Article Content
Abstract
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%.
Article Details
![Creative Commons License](http://i.creativecommons.org/l/by-nc-sa/4.0/88x31.png)
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.
Authors who publish with this journal agree to the following terms:
a). Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Attribution-NonCommercial-ShareAlike 4.0 International that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
b). Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
c). Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).
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