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ANALISIS SIDIK JARI KROMATOGRAM Stevia rebaudiana SECARA HIERACHIAL CLUSTER ANALYSIS (HCA) DAN PRINCIPAL COMPONENT ANALYSIS (PCA) Martono, Yohanes; Riyanto, Sugeng; Martono, Sudibyo; Rohman, Abdul
Majalah Obat Tradisional Vol 21, No 1 (2016)
Publisher : Faculty of Pharmacy, Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/tradmedj.10722

Abstract

Bioaktivitas tanaman herbal sangat dipengaruhi oleh kandungan senyawa aktif dan konsistensinya. Stevia rebaudiana mengandung senyawa aktif diterpen glikosida yang berkhasiat untuk antidiabetes. Tujuan penelitian ini adalah mengembangkan metode analisis sidik jari tanaman herbal S. rebaudiana berdasarkan kromatogram High Performance Liquid Chromatography (HPLC). Sampel daun S. rebaudiana berasal dari 5 daerah, usia dan asal bibit yang berbeda. Sampel daun S. rebaudiana dianalisis dengan metode Reversed Phase High Performance Liquid Chromatography (RP-HPLC) isokratik. Analisis sidikjari kromatogram S. rebaudiana menggunakan metode kemometrika secara Hierarchical Cluster Analysis (HCA) dan Principal Component Analysis (PCA). Pemilihan peak penanda (marker) “common peak” dilakukan dengan melakukan uji HCA secara Cluster Observation pada waktu retensi setiap puncak yang muncul di setiap kromatogram sampel. Waktu retensi yang memberikan nilai similarity level lebih dari 0,90 digunakan sebagai “common peak”. Analisis HCA yang dilakukan memberikan 5 puncak “common peak” sebagai penanda kromatogram yaitu peak no 1, 2, 4, 6, dan 7 dan dikelompokkan menjadi tiga klaster utama. Analisis PCA yang dilakukan dioptimalkan dengan memasukkan data area puncak yang memiliki nilai N > 2000 yaitu puncak no 4, 6 dan 7. Hasil PCA yang diperoleh dapat mengklasifikasikan kromatogram berdasarkan asal bibit, tempat tumbuh dan usia daun. Metode analisis sidik jari yang dikembangkan dapat dijadikan alternatif metode untuk kontrol kualitas tanaman herbal S. rebaudiana berdasarkan karakteristik sistemik kandungan senyawa fikokimia.
Potency of Industrial Tea Waste: Comparison Between Green And Black Tea Industrial Wastes as UV Filter for Sunscreen Martono, Yohanes
Indonesian Journal of Cancer Chemoprevention Vol 1, No 1 (2010)
Publisher : Indonesian Research Gateway

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Abstract

Indonesia is one of ten biggest countries that produce tea for world. It makes Indonesia produce various tea products. Every tea production process produces the large quantity of industrial tea wastes every day. Our previous research showed that industrial tea wastes still have antioxidant activity. It means that industrial tea wastes contained of phenolic compounds which can be used as UV filter for sunscreen. This research compared antioxidant activity, total phenolic contents and UV filter effectiveness  between green and black industrial tea wastes. Antioxidant activity were analyzed by reducing power and DPPH  method, total phenolic contents of tea wastes extract were analyzed  using Folin-Ciocalteu assay, while UV filter effectiveness were assessed by UV spectra and absorbance of each tea wastes extract related to its concentration in order to yield maximum protection. The results showed that although green tea waste extract had higher antioxidant activity but adversely, black tea had higher total phenolic contents. UV filter effectiveness is affected by polyphenols content in substances, so it suggested that black tea waste extract is more potential than green tea waste extract as photoprotection substance.
Pemberdayaan Petani Singkong Desa Kendel, Kecamatan Kemusu, Kabupaten Boyolali melalui Sentuhan Fortifikasi-Fermentasi Singkong Hartini, Sri; Martono, Yohanes
Agrokreatif Jurnal Ilmiah Pengabdian kepada Masyarakat Vol 1, No 1 (2015): Agrokreatif Jurnal Ilmiah Pengabdian kepada Masyarakat
Publisher : Institut Pertanian Bogor

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Abstract

Kendel village located in the Kemusu District, Boyolali. Cassava is one of the agricultural products in the region with 2.110 tons harvested cassava to the breadth of 212.4133 ha per year. Cassava plants is favorite plants for the population because it is relatively easy to cultivate and as a management area strategy because of limited water availability on the hilly areas. Up to now, post harvest processing of cassava is still using conventional methods and on sale in the form of raw cassava with its peels, and gaplek (dried cassava). While the cooked one are also served with simple ways, such as boiled cassava, fried cassava, roasted cassava, and lemet cassava. One of the constraints on the creation of cassava processing is limited diversification knowledge of the society Kendel as well as people do not yet understand the fortification of cassava. Close cooperation between Satya Wacana Christian University, Trukajaya Foundation, and Cassava Farmers Group realized empowerment, sortification, post harvest, advanced products, and increasing on economic value. The collabor ation was empowering community by using of participatory rural appraisal (PRA) method. Training have been done on post harvest handling, fortified cassava flour (mocaf), the manufacture of various products of mocaf flour, marketing training for products, monitoring and evaluation of processed products, and networking with other partners to develop programs based on local wisdom potential and organic as well. Various products have been produced from cassava flour fortified, processed products such as brownies, banana cake, cupcake, or cookies. Mocaf fortified flour sales price was Rp 8.000/kg.
AKTIVITAS ANTIOKSIDAN DAUN GINSENG JAWA (Talinum paniculatum Gaertn) Antioxidant Activity of Javanese Ginseng (Talinum paniculatum Gaertn) Leaves Lestario, Lydia Ninan; Christian, Anggelia Essi; Martono, Yohanes
Jurnal Agritech Vol 29, No 02 (2009)
Publisher : Jurnal Agritech

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Abstract

A research on antioxidant activity of Javanese Ginseng (Talinum paniculatum Gaertn) leaves has been carried out. The aims of this research were to compare the antioxidant activity and phenolic content of javanese ginseng leaves extract- ed by various kinds of solvent and to compare the antioxidant activity and phenolic content between and dried javanese ginseng leaves extracted by hot water. The antioxidant activity were measured by free radical scavenging method with DPPH, ferric thiocyanate method, and reducing power method, whereas the phenolic content was measured by Folin– Ciocalteu. The results showed that the highest antioxidant activity of javanese ginseng was found in ethanol extract: 43.78 %, IC = 273.13 µg/ml (by free radical scavenging method with DPPH); 93.17 % (by ferric thiocyanate meth- od); and 0.7022 meq K Fe(CN) /g extract (by reducing power method). The highest phenolic content was also found in4 6ethanol extract: 171.03 mg/g extract. The antioxidant activity of hot water extract of fresh leaves was higher than from dried leaves: 55.03 %, IC = 181.15 µg/ml (by method DPPH); 85.59 % (by ferric thiocyanate method); whereas the antioxidant activity by reducing power method and the phenolic content of the hot water extract of javanese ginseng leaves found in dried leaves were : 0.8078 meq K Fe(CN) /g extract and  116.11 mg/g extract respectively.ABSTRAKPenelitian tentang aktivitas antioksidan daun Ginseng Jawa (T. paniculatum Gaertn) telah dilakukan. Tujuan dari penelitian ini adalah untuk membandingkan aktivitas antioksidan dan kadar fenolik daun ginseng jawa yang diekstrak dengan beberapa jenis pelarut, dan membandingkan aktivitas antioksidan dan kadar fenolik ekstrak air panas daun ginseng jawa segar dan ekstrak air panas daun ginseng jawa yang dikeringkan. Aktivitas antioksidan diukur dengan metode penangkapan radikal bebas dengan DPPH, metode feri-tiosianat, dan metode kemampuan mereduksi; sedang- kan kadar fenolik diukur dengan metode Folin – Ciocalteu. Hasil penelitian menunjukkan bahwa aktivitas antioksidan tertinggi dari daun ginseng jawa diperoleh dari ekstrak etanol, yaitu sebesar 43,78 %, dengan nilai IC = 273,13 µg/ ml (metode penangkapan radikal bebas DPPH); 93,17 % (metode feri tiosianat) ;dan 0,7022 mek K Fe(CN) /g ekstrak4 6(metode kemampuan mereduksi). Kadar fenolik tertinggi juga terdapat dalam ekstrak etanol sebesar 171,03 mg/g ek-strak. Aktivitas antioksidan ekstrak air panas daun ginseng jawa segar lebih tinggi daripada ekstrak air panas daun kering, yaitu sebesar 55,03 % dan IC = 181,15 µg/ml (metode DPPH); 85,59 % (metode feri-tiosianat); sedangkan aktivitas antioksidan dengan metode kemampuan mereduksi dan kadar fenolik,  ekstrak air panas daun ginseng jawa kering lebih tinggi daripada ekstrak daun segar, yaitu berturut-turut sebesar 0,8078 mek K Fe(CN) /g ekstrak dan4 6116,11 mg/g ekstrak.
Analisis Kromatografi Cair Kinerja Tinggi untuk Penetapan Kadar Asam Galat, Kafein dan Epigalokatekin Galat pada Beberapa Produk Teh Celup Martono, Yohanes; Martono, Sudibyo
Agritech Vol 32, No 4 (2012)
Publisher : Faculty of Agricultural Technology, Universitas Gadjah Mada, Yogyakarta, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/agritech.9578

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A rapid and accurate High Performance Liquid Chromatography (HPLC) method for determination of gallic acid, caffein, and epigallocatechin gallate (EGCG) with an isocratic mobile phase elution was developed. HPLC parameters consisted of a C18 reversed-phase column (Eurosphere C18, 250 × 4.6 mm i.d., 5μm), mobile phase composed of 0.1% ortho phosporic acid : water : acetonitrile : methanol (14 : 7 : 3 : 1 v/v/v/v) pH = 4.00, flow rate 1.2 mL/min and was detected by UV detector at 280 nm. The validation of this method was confirmed from selectivity, linearity, accuracy and precision, limit of detection and limit of quantification. The method developed has proved to comply with validation requirements and has been applied to analyze various tea bag products (green and black tea). In green tea, the results showed that the highest amount of constituent was EGCG followed by caffein and gallic acid, whereas in black tea, EGCG can not be measured. ABSTRAK Metode Kromatografi Cair Kinerja Tinggi (KCKT) yang cepat dan akurat untuk penetapan kadar asam galat, kafein dan epigalokatekin galat (EGCG) dengan sistem elusi fase gerak secara isokratik telah dikembangkan. Sistem KCKT terdiri atas kolom fase terbalik C18 (Eurosphere C18, 250 × 4,6 mm i.d., 5μm), fase gerak campuran asam orto-fosfat 0,1% : air : asetonitril : metanol (14 : 7 : 3 : 1 v/v/v/v) pH = 4,00 dengan kecepatan alir 1,2 mL/min serta dideteksi pada UV 280 nm. Validasi metode ini dikonfirmasi terhadap selektivitas, linearitas, akurasi, presisi, batas deteksi dan batas kuantitasi. Metode yang dikembangkan dapat memenuhi syarat-syarat validasi metode dan telah digunakan untuk menganalisis berbagai produk teh celup (teh hijau dan hitam). Pada teh hijau, hasilnya menunjukkan bahwa kandungan terbesar adalah EGCG diikuti kafein dan asam galat sedangkan pada teh hitam, EGCG tidak terkuantitasi.
Protoporphyrin IX Extraction from Quail Eggshell (Cortunix cortunix) and Its Complexity with Zn(II) Laksono, A. Rindang Anggit; Martono, Yohanes; Riyanto, Cucun Alep
Eksakta: Jurnal Ilmu-Ilmu MIPA VOLUME 18, ISSUE 2, August 2018
Publisher : Fakultas Matematika dan Ilmu Pengetahuan Alam

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20885/eksakta.vol18.iss2.art7

Abstract

The aim of this research is to find the optimal condition for  Zn(II) complexation with protoporphyrin IX which is a natural coloring pigment on quail eggshell. The concentration of protoporphyrin IX extract is  0.01% (w / w) of the dried extract. The results of the complex are analyzed using Response Surface Methodology (RSM). The design used was central composite design with 3 factor variables X1 = pH (2; 4; 6), X2 = ratios (1: 3,1: 4,1: 5) and X3 = time (30 , 60, 90 ). The modeling used is linear and shows that the pH, ratio, and time factor have an influence on the complexation. This model gives the absorbance equation Y = 2.12506 - 0.049856X1 - 020316X2 - 0,00409857X3 and gives treatment in the most optimum complexation of pH 2; ratio of 1: 3; and time for 30 minutes with the absorbance of 1,293 and the desirability value of 0.825.
PENAMBAHAN GULA DAPAT MENINGKATKAN STABILITAS WARNA EKSTRAK ANTOSIANIN BUAH MURBEI HITAM YANG TERPAPAR CAHAYA FLUORESENS Soeroso, Erlien Giovani; Lestario, Lydia Ninan; Martono, Yohanes
Jurnal Teknologi dan Industri Pangan Vol 28, No 1 (2017): Jurnal Teknologi dan Industri Pangan
Publisher : Departemen Ilmu dan Teknologi Pangan, IPB Indonesia bekerjasama dengan PATPI

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (407.725 KB) | DOI: 10.6066/jtip.2017.28.1.62

Abstract

 Anthocyanins are red to purple pigments and one of potent safe and natural food colorants which possess superior antioxidant activity. However, the stability of anthocyanins depends on various factors, including structure and concentrations, pH, temperatures, light, co-pigment, ascorbic acid, enzymes, metal ions, sugars, protein, sulfur dioxide. The aims of this research were to evaluate the effects of sugar addition on the stability of anthocyanin of black mulberry under different fluorescent light intensities and to determine the appropriate concentration of sugar that could increase black mulberry (Morus nigra L.) anthocyanin stability. Fluorescent lamp at 3370, 4210, and 8820 lux (18, 23, and 32 watt, respectively) were used to illuminate black mulberry fruit extract (10.09 g/L) following 20, 40, 60% (w/v) of sugar addition. Determination of anthocyanin color stability in extract of black mulberry fruit was perfomed using degradation kinetics modeling to obtain the appropriate order. The concentration rate (k) and half life (t1/2) were then calculated based on subsequent order. The results showed that 20% sugar addition enhance the stability of black mulberry anthocyanin extract as compared to that of control (without sugar addition) as indicated by half life increment from 170.826 to 183.733 h. On the other hand, anthocyanin stability decreased to 166.618 and 146.235 h half life with further sugar addition to 40 and 60%, respectively. Therefore, 20% sugar addition to black mulberry extract could improve its anthocyanin stability.
Pengaruh Fermentasi terhadap Kandungan Protein dan Asam Amino pada Tepung Gaplek yang Difortifikasi Tepung Kedelai (Glycine max (L)) Martono, Yohanes; Danriani, Lucia Devi; Hartini, Sri
Agritech Vol 36, No 1 (2016)
Publisher : Faculty of Agricultural Technology, Universitas Gadjah Mada, Yogyakarta, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/agritech.10684

Abstract

Enhancement of proteins and enrichment of amino acids of dried cassava flour can be carried out by fermentation of dried cassava and soy-flour mixture. The purposes of this study were to compare soluble proteins content and to identify amino acids in fortified dried-cassava flour. The methods involved were making fortified dried-cassava flour with ratio of 25 g (soy-flour) and 5 g (yeasts) of 100 grams dried-cassava and fermentation of the mixture for 40 h, measuring soluble proteins content by biuret assay and identifying amino acids using Thin Layer Chromatography (TLC) and High Performance Liquid Chromatography (HPLC). Research design used Randomized Completely Block Design (RCBD). Treatments applied were unfortified dried cassava flour at 0 hour (G0); 20 hour (G20); 40 hour (G40) and fortified dried cassava flour at 0 hour (GF0); 20 hour (GF20) and 40 hour (GF40). As group was analyisis time. Data of soluble protein content were analyzed using ANNOVA. The Honestly Significant Differences (HSD) test at 5% level of significance was used to compare the treatments mean. The results showed that the soluble proteins content of 40 hours fermentation was higher than of 20 hours fermentation and the latter was higher than 0 hour fermentation, with values of 22.86%, 20.96%, and 18.70%, respectively. Based on TLC and HPLC identification, the amino acids in fortified dried-cassava flour, were aspartate, glumatate, serine, histidine, glicine, arginine, alanine, tyrosine, methionine, valine, isoleusine, leusine, and lysine.ABSTRAKPeningkatan kadar protein dan pengayaan asam amino dalam tepung gaplek dapat dilakukan dengan cara fortifikasi tepung gaplek dengan tepung kedelai melalui fermentasi. Tujuan dari penelitian ini adalah membandingkan kandungan protein  terlarut  antara  tepung  gaplek  tidak  terfortifikasi dan  terfortifikasi tepung  kedelai  pada  beberapa  waktu fermentasi dan mengidentifikasi asam amino yang terkandung dalam tepung gaplek dan tepung gaplek terfortifikasi. Metode penelitian meliputi pembuatan tepung gaplek terfortifikasi secara fermentasi dengan rasio penambahan tepung kedelai 25 g dan kapang 5 g dari 100 g gaplek yang kemudian difermentasi selama 40 jam. Kadar protein ditentukan menggunakan metode Biuret sedangkan identifikasi asam amino menggunakan metode Kromatografi Lapis Tipis (KLT) dan Kromatografi Cair Kinerja Tinggi (KCKT). Rancangan percobaan penelitian menggunakan Rancangan Acak Kelompok (RAK). Sebagai perlakuan adalah gaplek kontrol (tidak terfortifikasi tepung kedelai) pada jam ke-0 (G0); 20 jam (G20); 40 jam (G40) dan gaplek terfortifikasi pada jam ke-0 (GF0); 20 jam (GF20); dan 40 jam (GF40). Sebagai kelompok adalah waktu analisis. Data rata-rata kadar protein terlarut dibandingkan dengan uji Beda Nyata Jujur (BNJ) dengan tingkat kebermaknaan 5%. Hasil penelitian ini menunjukkan bahwa kadar protein terlarut pada fermentasi selama 40 jam lebih tinggi dibandingkan 20 jam dan 0 jam, dengan nilai 22,68%, 20,96%, dan 18,70% secara berurutan. Berdasarkan hasil identifikasi KLT dan KCKT, asam amino dalam tepung gaplek terfortifikasi adalah aspartat, glutamat, serin, histidin, glisin, arginin, alanin, tirosin, metionin, valin, isoleusin, dan lisin.
Determination of fluoride content in toothpaste using spectrophotometry Hastuti, Susanti Pudji; Lestari, Devinta; Martono, Yohanes
Dental Journal (Majalah Kedokteran Gigi) Vol 46, No 3 (2013): (September 2013)
Publisher : Faculty of Dental Medicine, Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20473/j.djmkg.v46.i3.p124-129

Abstract

Background: Intake excessive fluoride in children’s teeth are generally marked with white and brown patches. Excessive fluoride of more than 4.0 mg/L can cause a person suffering from poisoning, fragility of the bones (osteoporosis), liver and kidney damage. Knowledge about the spectrophotometry for determination method of fluoride content in commercially available toothpaste is very few. Purpose: The purposes of study were to examine the suitable method for fluoride extraction and to determine out the accuracy, precision, linearity, and stability of the measurement method of fluoride content in toothpaste. Methods: The suitable F extraction method was determined by the comparison among 3 methods of extraction; e.g. the dried samples were immersed in (1) distilled water, (2) 96% HCl, and (3) 96% HNO3; and the validation methods of measurement were the maximum wavelength, standart curve, accuracy test, precision test, and stability test. Results: Result showed that the fluoride extraction by using the concentrated HNO3 was found to have the highest levels of fluoride, followed by hydrochloric acid dissolution (HCl) and distilled water, while the method of validation showed that SPADNS revealed the acceptable accuracy. Precision has the RSD ≤ 2.00%. Furthermore the stability test result showed that the measurement of fluoride less than 2 hours was still reliable. Conclusion: The study suggested that the best result of fluoride extraction from toothpaste could be gained by using concentrate HNO3, and the spectrophotometer (UV-Vis Mini Shimadzu U-1240) and SPADNS have the acceptable accuracy.Latar Belakang: Pemasukan fluoride yang berlebihan pada gigi anak ditandai dengan bercak putih dan coklat. Fluoride lebih dari 4.0 mg / L dapat menyebabkan seseorang menderita keracunan , kerapuhan tulang (osteoporosis), kerusakan hati dan ginjal. Pengetahuan tentang spektrofotometri untuk metode penentuan kadar fluoride dalam pasta gigi yang tersedia secara komersial sangat sedikit . Tujuan: Penelitian ini bertujuan meneliti metode yang tepat untuk mengektrak kandungan fluoride dan mengukur akurasi, presisi, linearitas dan stabilitas pengukuran kandungan fluoride pada pasta gigi. Metode: Metode ekstrak yang tepat ditentukan dengan membandingkan 3 metode, yaitu dengan perendamam sampel kering dalam (1) air destilasi, (2) HCl 96%, dan (3) HNO3 96%; dan validasi metode yang memperhitungkan panjang gelombang, kurva standar, tes akurasi, presisi, dan stabilitas. Hasil: Hasil menunjukkan bahwa metode ekstrasi fluoride pada pasta gigi dengan menggunakan metode HNO3 didapatkan level fluoride tertinggi, diikuti dengan metode HCl dan air destilasi. Hasil validasi metoda menunjukkan bahwa penggunaan SPADNS akurasinya dapat diterima. Presisi mempunyai RSD ≤2,00%. Pada tes stabilitas didapatkan hasil bahwa pengukuran kadar fluorida dalam waktu tidak lebih dari 2 jam masih dapat dilakukan. Simpulan: Penelitian ini menunjukkan bahwa hasil terbaik ekstraksi fluoride dari pasta gigi dapat diperoleh dengan menggunakan HNO3, dan spectrophotometer (UV-Vis Mini Shimadzu U-1240) dan SPADNS memiliki pengukuran yang akurat.
ISOLASI DAN ANALISIS GENISTEIN DARI TEMPE BUSUK MENGGUNAKAN METODE KROMATOGRAFI KOLOM Soetjipto, Hartati; Martono, Yohanes; Yuniarti, Zulfa
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol 5, No 1 (2018): June 2018
Publisher : Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (8416.45 KB) | DOI: 10.29122/jbbi.v5i1.2860

Abstract

Isolation and Analysis of Genistein of Overripe Tempe using Column Chromatography MethodABSTRACTGenistein 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 ABSTRAKGenistein 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