Nghiên cứu định lượng các hoạt chất trong sinh khối thông đỏ bằng sắc ký lỏng hiệu năng cao

jjghien CLFU - Ky thuat

^hien ciru dinh lucrng cac hoat chat r

trong sinh khoi thong do > r

bang sac ky long hieu nang cao

Vu Binh Dirong^, Nguyen Van Long^,

Pham Thi Thanh Ha', Le Thi Thien*, Dao Van Don^

Trudng dgi hoc Dugc Hd Ngi; ^ Hge Vien Qudn Y

Dat van d e

Thong do Viet Nam {Taxus wallichiana) Id

dugc lieu quy phan b i chii yiu tai eae huyen

Dire Trgng, Dan Duang, Lae Duang vd thdnh

phi Od Lat, tinh Ldm Ding. Theo ede tdi lieu

nghien ciru cua cac nhd khoa hge, trong thong

do Viet Nam co chua cac boat chat co tae dung

ipc chi t i bdo ung thu nhu: paelitaxel (taxol),

cephalomannin hoac cae ehit cd the ban ting

hgp ra cae thuic diiu tri ung thu nhu baeeatin

III, 10 deacetyl baeeatin III, brevifoliol...

Hien nay, tai Trung tam Nghien euu ung dung

Sinh - Y- Duge (Hpc vien Quan Y) da budc diu

thdnh cdng trong viec nu6i ciy tac sinh khii t i bdp

thong dd trong phong thf nghiem, dang tang din

qui mo san xuat cdng nghiep .

De gop phin ddnh gid chit lugng eua sinh

khii th6ng do san xuit tai Hge vien Quan y,

chiing tdi tiin hdnh d i tai ndy vdi mue tieu xay

dung qui trinh djnh lugng eae boat chit chinh

(paelitaxel vd baeeatin III) trong sinh khii thong

do va ung dung qui trinh da xdy dung trong

danh gia ehit lugng sinh khii thdng dd.

Nguyen lieu va thiet bj

Sinh khii th6ng do (Hge vien Qudn y eung

cip, L6 SX 050909; 060909; 070909).

He thing HPLC Alliance Waters 2695D,

autosampler, buing gia nhiet cot. Detector PDA,

Phin mim xu' ly s i lieu Empovi/er® 2, cpt Fortis

Phenyl (250 x 4,6 mm, 5 pm) va Cpt Luna L43

(250 x 4,6 mm; 5 pm); Can phan tieh Sartorius

(dp chfnh xdc 0,1 mg), mdy ly tam lanh

UNIVERSAL 320 (Hettich - Due), may khuiy tu

gia nhiet RCT Basic, IKA® USA, mdy lic MSI

Minishaker, IKA® USA, tu lanh am sau DF8514

(llshinlab - Han Quicj, nhiet dp am ti i da -

80°C, micropipet_1-10iil, lO-IOOiil, lOO-IOOOiil,

bp phan lpc: phiu Ipc Buchner, bp phan Ipc

miu, mang Ipe 0,2Mm; 0,45Mm.

TAP CHI DUQC HOC - 9/2010 (SO 413 NAM 50)

Cdc ehit ehuin paelitaxel va baeeatin III

(Sigma - My).

ThLPC nghiem va k§t qua

Phipcng phap xd" ly mlu

Sau khi tham khao m6t s i tai lieu ' va

lua chpn cae diiu kien hien eo, chung toi tiin

hdnh khao sat phuang phdp xu' ly miu vdi cdc

diiu kien sau:

- Chiit long - Idng vdi hin hgp dung mdi

methanol - dieloromethan (3:1, tt/tt), ehiit nhiiu

lin, rii phan tich ngay bing HPLC.

- Chiit long - rin vdi dung m6i ehiit

methanol, chiit nhiiu lin, sau do tinh ch i bing

qua cot chiit pha rin trudc khi chay HPLC

Kit qua si c ky d i cua 2 phuang phdp XLP ly

miu thi hien d hinh la va lb eho thiy: phuang

phap ehiit long - rin eo qua ept ehiit pha rin thi

pie boat chit paelitaxel va baeeatin III tdeh ro

rang ra khoi hoat chit, ding thdi kha nang chiit

boat chit tit han phuang phap ehiit long - long.

Vi vay, ehung toi chpn phuang phap xu- ly mdu

nay eho nghien euu tiip theo.

Liy 200 mg b6t mjn sinh khii th6ng dd chiit

sieu am bing dung mdi methanol (12 ml x 3 lin),

ly tam 3000 vong/phiit liy phin djch trong phfa

tren, bic hai methanol tdi khi con 10 ml, bo sung

them 10 ml natri clorid 5% vd chiit bing dung mdi

dieloromethan (10ml x 4 lin). Djch ehiit

dieloromethan dugc Ipc qua 3g natri sulfat khan,

djch Igc dugc bic hai tdi cdn. Hoa tan ein vdi 2

ml dielcromethan roi chuyen vao cpt silica gel Ig.

Rira cpt vdi dieloromethan (2x2ml) sau d6 vdi

aceton - dieloromethan (4:96, 2x5ml), bo djch ru-a.

Rua giai vdi aceton - dieloromethan (50:50,

2x5ml) gop nude rua, bic hai tdi ein (N2, 35°C),

boa tan cin trong 1ml methanol trudc khi dua vao

phan tich HPLC.

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