ISOLASI DAN IDENTIFIKASI SENYAWA AKTIF ANTI TUMOR DARI EKSTRAK AIR BENALU TEH (Scumsla oortiana)

Abstract

Earlier studies on the anti tumor acitivity of parasitic tea plants S. oortiana has shown that the stem extracts is stronger than the leave extracts in modulating the sensitivity of fibrosarcoma to TNFa. Based on this finding further studies was undertaken to isolate and identified the active constituents in the extract which exert their anti tumor activities. The water extracts of S. oortiana was further fractionated by column chromatography on Silica gel and eluted by Chloroform-Metanol. This fractionation produced 15 fractions. Based on DPPH and fibrosarcom inhibition tests guide , some fractions were further purified through Si02 column eluted with Methanol-water and Sephadex LH-20 eluted with Methanol-water. This is followed by High Pressure Liquid Chromatography analysis on ODS column with solvent KH2PO4-acetonitril which produced pure isolates designated as A, B, C and D. Ultra Violet, Infra Red, Proton and Carbon Nuclear Magnetic Resonance, and Gas Chromatography-Mass Spectrum Analysis of the four isolates reveals that A is caffeine, B is Catechin, C is Phytol, and D is Glicosidic Flavonoid. Analysis for an anti tumor activity by DPPH test for Catechin and Phytol showed ICso 82,4 ppm and 88,77 ppm respectively. An anti tumor test on fibrosarcoma showed that Catechin, Phytol, and Flavonoid can lyse directly the tumor cells where Phytol showed the greatest activity. Ekstraksi S. oortiana dengan air kemudian dilanjutican fraksinasi dengan analisis kromatografi kolom (silica gel; CHC13-Me0H) memberikan 15 fraksi. Dengan panduan bioesai anti tumor, beberapa fraksi yang memberikan reaksi positif dilanjutkan pemurnian dengan analisis kromatografi kolom (Si02, CHC13-Me0H-H20; Sephadex LH-20, Me0H-H20) dan analisis kromatografi cair kinetja tinggi (KCKT, ODS, KH2PO4-aetonitril) memberikan senyawa isolat mumi A, B, C dan D. Hasil interpretasi spectra ultra violet (UV), infra merah (IR), Resonansi magnetic Intl (RMI proton dan karbon) dan sepktrometri massa (GC-MS) senyawa tersebut ditetapkan sebagai senyawa kafein (A), senyawa B sebagai catechin, senyawa C sebagai Phytol, dan D sebagai senyawa flavonoid glikosida. Senyawa-senyawa mumi Catechin dan Phytol menunjukkan aktifitas anti tumor melalui uji DPITI dengan IC50 berturut-turut yaitu 82,4 ppm dan 88,77 ppm. Uji aktifitas anti tumor melalui uji terhadap sel fibrosarcoma menunjuklcan aktifitas lysis oleh masing-masing senyawa catechin, phytol, maupun flavonoid dimana senyawa Phytol beketja dalam waktu yang lebih singlcat dibandingkan Catechin dan Flavonoid.