Cara Pembuatan SimplisiaPemanenanPengumpulanSortasi basahPencucianPerajanganPengeringanSortasi keringPengawetanPengemasan

Pada saat pemanenan, pengumpulan, dan sortasi basah

Perhatikan garis-garis besar pemanenan.

Pencucian dengan air mengalir untuk mengilangkan debu serta pengotor yang melekat

Sesudah dicuci, bahan diangin-anginkan untuk mengurangi air yang menempel.

Pengeringan, dapat dilakukan diAlmari pengeringOvenSinar matahari

Pengemasan untuk mempertahankan mutu simplisia dalam penyimpanan.

Yang menjadi perhatian:Pedoman panenPengirisan bahan sebelum dikeringkan, untuk bahan yang berukuran besar.Pengeringan,* di dalam oven* di dalam almari pengering* di bawah sinar matahari

Isolasi & Identifikasi alkaloid dari Akar Pasak Bumi (Eurycoma longifolia Jack)

Nina Salamah (PFA/561)

Eurycoma longifolia Jack leaves and fruits Eurycoma longifolia Jack 6-years old plant and roots

Eurycoma longifolia JackAkar Pasak Bumi (Eurycoma longifolia Jack) merupakan family Simaroubaceae dikenal sebagai obat kuat (Tongkat Ali). Tanaman ini tumbuh di hutan Indonesia (Kalimantan) juga di Malaysia

Di Asia Tenggara digunakan untuk antimalaria, antipyretic, antiulcer, cytotoxic dan aphrodisiaka.

Eurycoma longifolia JackMenurut kardono ada 5 komponen aktif akar pasak bumi yang berpotensi sitotoksik yaitu: 4 merupakan gol alkaloid canthin-6-one (9-methoxycanthin-6-one, 9-methoxycanthin-6-one-Noxide, 9-hydroxycanthin-6-one, 9-hydroxycanthin-6-one-N-oxide) dan 1 golongan quassinoid (eurycomanon).Ke 5 senyawa diuji sitotoksiknya pada beberapa tipe sel kanker [breast, colon, fibrosarcoma, lung, melanoma, KB, and KB-V1 (a multi-drug resistant cell line derived from KB)] and murine lymphocytic leukemia (P-388)The canthin-6-ones 1-4 were found to be active with all cell lines tested except for the KB-V 1 cell line. Eurycomanone was inactive against murine lymphocytic leukemia (P-388) but was significantly active against the human cell lines tested.

Alkaloid canthin-6-one 9-methoxycanthin-6-one [1], 9-methoxycanthin-6-one-N-oxide [2],9-hydroxycanthin-6-one [3], and 9-hydroxycanthin-6-one-N-oxide [4]

RisetIsolasi senyawa (1) dan (3) untuk dikembangkan sebagai obat anti kankerDari strukturnya, kedua senyawa ini mempunyai sifat yang kurang polar. Perlu dipisahkan dari senyawa lain yang berbeda kepolarannya

Cara isolasi100 g serbuk akar kemudian perkolasi dengan MeOH ekstrak MeOH CC silica gel dengan fase gerak CHCL3:MeOH dalam berbagai perbandingan 9 fraksi (F002-F010) Fraksi 003 CC si gel ( CHCl3 :Me OH= 99:2) 9 methoksi canthin 6 oneFraksi 006 CC Si Gel ( CHCl3 : MeOH = 96:4) 9 hidroksi canthin 6 one

evaporasiMaserasi berulang dg etanol 50%UVFTIRGC/MSNMRSimplisia Pasak BumiMaseratCrude Ekstrakn-ButanolDietil eterPurifikasi dengan HPLC semi preparatif Kolom C18, 2 ml/menit, Deteksi UV 240 nmAcetonitril : air (3:7)eurycomalacton9-methoxycanthin-6-oneFase gerak

Proses KVCDiameter kolom = 5 cmBerat silika = 30 70 g(tinggi silika = 5 -6 cm)Kapasitas sampel = 5 gVolume eluen = 50 75 mLKunci pemisahan/pemurnian yang baik, pada pemilihan eluen (fase gerak).

PREPARASI KOLOM

9-methoxycanthin-6-one Identifiksi kualitatif:KLT dengan CHCl3-MeOH (98:2) as the solvent system, indicated further their co-identity (Rf O. 50)

Tabel 1.13C-nmr Assignments for Compounds 1compound 1 was directly comparable (mmp, uv, ir, 'H-nmr, 13C nmr, ms, co-tlc) toan isolate obtained earlier in this laboratory from the wood of S. multiflora, which wasassigned as 10-methoxy canthin-6-one using primarily low resolution 'H-nmr spectroscopy (14). Therefore, this S. multiflora isolate has now been reassigned as 9-methoxy canthin-6-one.

CarbonppmC-1C-2C-4C-5C-6C-8C-9C-10C-11C-12C-13C-14C-15C-16O-CH3115.59146.02139.92128.56159.61101.41163.20114.26123.47117.03132.02129.24131.16136.0456.03

9-hydroxycanthin-6-oneIdentifikasi kualitatifSi gel tlc migrationdata (Rf 0.40), using CHCl,-MeOH (95:5).

Tabel 2. 13C dan 1H-nmr Assignments for Compounds 3

CarbonppmC-1C-2C-4C-5C-6C-8C-9C-10C-11C-12C-13C-14C-15C-16O-CH3116.01145.98140.03128.04158.94102.98160.50114.00124.65115.57140.53129.88131.66135.01-

hidrogenppmH-1H-2H-4H-5H-8H-10H-11O-CH38.14(d, 5.0)8.76(d, 5.0)8.10(d, 10)6.96(d, 10)8.00(d, 2.5)7.00(dd, 9.2.5)8.17 (d, 9)-

13C-NMR dan 1H-NMR'H- and 13C-nmr resonances of this compound were quite comparable to those obtained for compound 1.After measuring the 'H-'H COSY and 'H-13C HETCOR nmr spectra of 3 and following comparison with published unambiguous 'H- and 13C-nmr assignments for canthin-6-one (17), the hydroxy group was tentatively placed at C-9. In a selective INEPT experiment (JCH= 8 Hz) performed with 3, irradiation of H-2 ( 8.76) enhanced the resonances of C-14 ( 129.88) and C-16 ( 135.03), while irradiation of H-11 ( 8.17) enhanced the resonances of C-14 ( 129.88), C-9 ( 160.50), and C-13 ( 140.53). The data obtained from a 'H-13C COLOC experiment also supported the placement of the hydroxy group at C-9. Final proof of the structure of 3 was obtained by methylation with CH2N2 to afford compound 1

9-methoxycanthin-6-one [coumpound1], 9-hydroxycanthin-6-one [coumpound3]Compound 3, yellow needles, mp 285-286" (dec); uv max (MeOH) (log E) 238 (sh, 4.20), 274 (4.20), 310 (4.05), 352 nm (4.15); (+ NaOH), 236 (sh, 4.25), 294 (4.15), 316 (sh, 4. 10), 355 (4.00),428 nm (3.90); ir v max (KBr) 3400-3600, 1700, 1663, 1653, 1636, 1570, 1559, 1457, 1355, 1320,1285, 1250, 1155, 1064 ,986,929, 850, 835, 635 cm-'; 'H nmr see Table 2; I3C nmr see Table 1; eims (70 eV) m/z [M]+ 236 (100), 224 (10), 209 (8),2 08 (50), 197 (24), 179 (10), 150(58), 123 (22), 118 (5); hreims 236.0590 (calcd for C14H8N202, 236.0585). When compound 3 (5 mg) was methylated with CH2N2 using the Diazald Kit (Aldrich, Milwaukee, Wisconsin), 4 mg of a product was obtained that was identical (mmp, 'H nmr, uv, ir, ms) to compound 1.