TIU CHUN KIM NGHIM ETOPOSIDE

THEO DC IN ANH 2013

*NH TNH: c th chn 1 trong 2 nhm nh tnh sau
- Nhm I:A, B
- Nhm II:C, D.

A. Gc quay cc ring: t -106o n -114o (cht khan).

Ho tan 50,0 mg trong mt hn hp ca 1 th tch CH3OH (TT) v 9
th tch ca CH2Cl2 (TT) v pha long thnh 10,0 mL vi cng dung mi.

B. Ph hng ngoi (IR): Ph hng ngoi ca ch phm phi ph hp vi

ph hng ngoi ca epotosid chun (C)

C. Sc k lp mng:
-Dung dch th: Ha tan 10 mg ch phm cn kim tra trong mt hn hp
ca 1 th tch CH3OH (TT) v 9 th tch ca CH2Cl2 (TT) v pha long
thnh 2 mL vi cng dung mi.
-Dung dch i chiu: Ha tan 10 mg etoposide chun (C) trong hn hp
CH3OH (TT) v 9 th tch ca CH2Cl2 (TT) v pha long thnh 2 ml vi
cng dung mi.
-Bn mng: silica gel H (TT)
-Pha ng: H2O (TT), CH3COOH bng (TT) , CH3COCH3 (TT), CH2Cl2
(TT) t l (1.5:8:20:100 V/V/V/V).
- Chm ring bit ln bn mng mi 5 l dung dch th v dung dch i
chiu thnh di 10 mm. Trin khai sc k ngay lp tc, cho dung mi i n
6/7 bn mng. Lm kh di mt lung kh m trong 5 pht. Pht hin bng
cch phun hn hp ca 1 th tch H2SO4 (TT) v 9 th tch C2H5OH (96%)
(TT). Sy 140oC trong 15 pht. Ngay lp tc bao bn mng bng a thy
tinh cng kch c. Kim tra di nh sng ban ngy.
- Kt qu: Di chnh trn sc k ca dung dch th tng t v v tr,
mu sc v kch c vi di chnh trn sc k ca dung dch i chiu.

D. Trong ng nghim, ha tan khong 5 mg trong 5 mL CH3COOH

bng (TT) v thm 0,05 mL dung dch FeCl3 (TT). Trn v thm cn
thn 2 mL H2SO4 (TT). Trnh trn 2 lp. yn khong 30 pht; thy
xut hin 1 vng nhn mu hng n nu mt ngn cch gia 2 lp
v lp trn c mu vng
*TH TINH KHIT:
- trong ca dung dch: Dung dch u trong sut (ph lc 2.2.1) v
khng c mu m hn dung dch i chiu Y6 hoc BY6 (ph lc 2.2.2,
phng php II). Ho tan 0. mg trong mt hn hp ca 1 th tch CH3OH
(TT) v 9 th tch ca CH2Cl2 (TT) v pha long thnh 2.0 mL vi cng
dung mi
-Gc quay cc ring: t -106o n -114o (cht khan). Ho tan 50,0 mg
trong mt hn hp ca 1 th tch CH3OH (TT) v 9 th tch ca CH2Cl2 (TT)
v pha long thnh 10,0 mL vi cng dung mi.
-Tp cht lin quan: phng php Sc k lp mng
+Hn hp dung mi: pha ng A, pha ng B (50:50 V/V).
+Dung dch th: (a) Ha tan 40 mg ch phm trong hn hp dung mi v
pha long thnh 10.0 mL vi hn hp dung mi.
+Dung dch th: (b) Ho tan 50.0 mg ch phm trong hn hp dung mi v
pha long thnh 50.0 ml vi hn hp dung mi.
+Dung dch i chiu (a) Pha long 1.0 ml dung dch th (a) n 100.0 mL
vi hn hp dung mi. Pha long 1.0 mL dung dch ny thnh 10.0 mL vi
hn hp dung mi.
+Dung dch i chiu (b) Ha tan 4 mg etoposide ph hp vi h thng
CRS (cha cc tp cht B, C, D, E, N v O) trong 1,0 mL hn hp dung mi.
+Dung dch i chiu (c) Ho tan 50.0 mg etoposide chun (C) trong dung
mi v pha long thnh 50.0 mL vi hn hp dung mi.
+Ct:
Kch thc: chiu di = 0.125 m, = 4.6 mm;
Pha tnh: silica gel octadecylsilyl (TT) (5 m);
Nhit : 40 C.
o Pha ng: Pha ng A: HCOOH khan (TT),, (C2H5)3N (TT),
nc (TT) t l (1:1:998 V/V/V);
o Pha ng B: HCOOH khan (TT),, (C2H5)3N (TT), CH3CN (TT)
t l (1:1:998 V/V/V);

+Tc dng: 1 mL / pht.

+Detector: Quang ph k bc sng 285 nm.
+Tim 10 l dung dch th (a) v dung dch i chiu (a) v (b).
+Xc nh cc tp cht: S dng sc k chun ca etoposide CRS v sc
k ca dung dch i chiu (b) xc nh cc nh do tp cht B, C, D,
E, N v O.
+Thi gian lu tng i: Etoposide c thi gian lu gi = khong 5 pht;
tp D = khong 0,4; Tp E = khong 0,8; Tp cht C = khong 1,1; Tp cht
B = khong 1,2; Tp cht N = khong 3,1; Tp cht O = khong 4,2.
+Tnh ph hp ca h thng: Dung dch i chiu (b)
Peak-to-valley ratio: ti thiu 2.0, trong Hp = chiu cao trn ng
nn ca nh do tp cht C v Hv = chiu cao trn ng nn ca im thp
nht ca ng cong tch nh ny t nh do etoposide; V ti thiu 3,0,
trong Hp = chiu cao trn ng nn ca nh do tp cht B v Hv =
chiu cao trn ng nn ca im thp nht ca ng cong tch nh ny
khi nh do tp cht C.
+Cc gii hn:
o h s hiu chnh: tnh hm lng, nhn din tch peak tp cht O
vi 1.7;
o tp cht B, C, D, E, N: i vi mi tp cht, khng ln hn hai ln
din tch pic chnh trn sc k ca dung dch i chiu (a) (0,2%);
o tp cht O: khng ln hn 1,5 ln din tch pic chnh trn sc k
ca dung dch i chiu (a) (0,15%);
o tp cht khng xc nh: i vi mi tp cht, khng ln hn din tch
pic chnh trn sc k ca dung dch i chiu (a) (0,10%);
o Tng: ti a 10 ln din tch pic chnh trn sc k ca dung dch
i chiu (a) (1,0 phn trm);
o gii hn b qua: 0,5 ln din tch pic chnh trn sc k ca dung
dch i chiu (a) (0,05%); B qua cc peak ca dung mi.
-Kim loi nng: nng ti a 20 ppm
0,5 g ph hp vi php th G. Dng 2 ml dung dch ch mu 10 phn
triu (TT) chun b mu i chiu
-Nc: Ti a 6,0%. Dng 0.250 g ch phm.
-Tro sulfate: Ti a 0,1%. Dng 1.0 g ch phm.

*NH LNG: bng pp sc k lng (nh m t trong cc th nghim

xc nh tp lin quan)
-Mu tim: Dung dch th (b) v dung dch i chiu (c).
-Tnh ph hp ca h thng: lp li: lch chun tng i ti a l
1,0% sau 6 ln tim dung dch i chiu (c).
 -Tnh hm lng phn trm ca Epotoside t ni dung k khai ca CRS
etoposide
TP CHT: Cc tp xc nh:B, C, D, E, N, O.

C. (5R,5aR,8aR,9S)-9-[[4,6-O-
[(R)-ethylidene]--D-
glucopyranosyl]oxy]-5-(4-
hydroxy-3,5-dimethoxyphenyl)-
5,8,8a,9-
tetrahydroisobenzofuro[5,6-f][1,
3]benzodioxol-6(5aH)-one (-
etoposide)

B. (5R,5aS,8aR,9S)-9-[[4,6-O-[(R)-ethylidene]--D-
glucopyranosyl]oxy]-5-(4- hydroxy-3,5-dimethoxyphenyl)-
5,8,8a,9-tetrahydroisobenzofuro[5,6-f][1, 3]benzodioxol-
6(5aH)-one (picroethylidene-lignan P; cis-etoposide)

N. (5R,5aR,8aR,9S)-9-[[4,6-O-[(R)-ethylidene]--D-
glucopyranosyl]oxy]-5-[4-[[(5R, 5aR,8aR,9S)-5-(4-
hydroxy-3,5-dimethoxyphenyl)-6-oxo-5,5a,6,8,8a,9-
hexahydroisobenzofuro[5,6-f][1,3]benzodioxol-9-
yl]oxy]3,5-dimethoxyphenyl]-5,8, 8a,9-
D. (5R,5aR,8aR,9S)-9-(-D-glucopyranosyloxy)-
tetrahydroisobenzofuro-[5,6-f][1,3]benzodioxol-
5-(4-hydroxy-3,5-dimethoxyphenyl)- 5,8,8a,9- 6(5aH)-one.
tetrahydroisobenzofuro-[5,6-f][1,3]benzodioxol-
6(5aH)-one (lignan P),
E. (5R,5aR,8aR,9S)-9-hydroxy-5-(4-hydroxy-
3,5-dimethoxyphenyl)-5,8,8a,9-
tetrahydroisobenzofuro[5,6-f][1,3]benzodioxol-
6(5aH)-one (4- desmethylepipodophyllotoxin)

O. (5R,5aR,8aR,9S)-9-[[2,3-bis-O-(dichloroacetyl)-4,6-O-
[(S)-ethylidene]--L- glucopyranosyl]oxy]-5-(4-hydroxy-3,5-
dimethoxyphenyl)-5,8,8a,9- tetrahydroisobenzofuro[5,6-f]
[1,3]benzodioxol-6(5aH)-one,