Cara Pembuatan Simplisia

1. Pemanenan
2. Pengumpulan
3. Sortasi basah
4. Pencucian
5. Perajangan
6. Pengeringan
7. Sortasi kering
8. Pengawetan
9. Pengemasan

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 di
1. Almari pengering
2. Oven
3. Sinar matahari
Pengemasan untuk mempertahankan mutu
simplisia dalam penyimpanan.
Yang menjadi perhatian:
1. Pedoman panen
2. Pengirisan bahan sebelum dikeringkan,
untuk bahan yang berukuran besar.
3. 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 Jack
Akar 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 Jack
Menurut 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]
Riset
Isolasi senyawa (1) dan
(3) untuk dikembangkan
sebagai obat anti kanker
Dari strukturnya, kedua
senyawa ini mempunyai
sifat yang kurang polar.
Perlu dipisahkan dari
senyawa lain yang
berbeda kepolarannya
Cara isolasi
100 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 one
Fraksi 006 CC Si Gel ( CHCl3 : MeOH =
96:4) 9 hidroksi canthin 6 one
evaporasi
Maserasi berulang dg etanol 50%
UV
FTIR
GC/MS
NMR
Simplisia Pasak Bumi
Maserat
Crude Ekstrak
n-Butanol
Dietil eter
Kolom Kromatografi
800 ml n-heksane:metanol
7:3 1:1 3:7
Purifikasi dengan HPLC semi preparatif Kolom C18,
2 ml/menit, Deteksi UV 240 nm
Acetonitril : air (3:7)
eurycomalacton 9-methoxycanthin-6-one
Fase gerak
Proses KVC
Diameter kolom = 5 cm
Berat silika = 30 70 g
(tinggi silika = 5 -6 cm)
Kapasitas sampel = 5 g
Volume eluen = 50 75 mL
Kunci
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)
N
N
H
3
CO
O
senyawa 1
9-methoxy-6H-indolo[3,2,1-ij][1,5]naphthyridin-6-one
Tabel 1.
13
C-nmr Assignments for
Compounds 1
compound 1 was directly
comparable (mmp, uv, ir, 'H-
nmr, 13C nmr, ms, co-tlc) to
an isolate obtained earlier in
this laboratory from the wood
of S. multiflora, which was
assigned 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.
Carbon ppm
C-1
C-2
C-4
C-5
C-6
C-8
C-9
C-10
C-11
C-12
C-13
C-14
C-15
C-16
O-CH3

115.59
146.02
139.92
128.56
159.61
101.41
163.20
114.26
123.47
117.03
132.02
129.24
131.16
136.04
56.03
9-hydroxycanthin-6-one
Identifikasi kualitatif
Si gel tlc migration
data (Rf 0.40), using
CHCl,-MeOH (95:5).
N
N
HO
O
senyawa 3
9-hydroxy-6H-indolo[3,2,1-ij][1,5]naphthyridin-6-one
Tabel 2.
13
C dan
1
H-nmr
Assignments for Compounds 3
Carbon ppm
C-1
C-2
C-4
C-5
C-6
C-8
C-9
C-10
C-11
C-12
C-13
C-14
C-15
C-16
O-CH3
116.01
145.98
140.03
128.04
158.94
102.98
160.50
114.00
124.65
115.57
140.53
129.88
131.66
135.01
-
hidrogen ppm
H-1
H-2
H-4
H-5
H-8
H-10
H-11
O-CH3
8.14(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)
-
13
C-NMR dan
1
H-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 (J
CH
= 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
CH
2
N
2
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
C
14
H
8
N
2
0
2
, 236.0585). When compound 3 (5 mg) was methylated
with CH
2
N
2
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.