• Nghien CIFU - Ky thuat
Xay dyng quy trinh
dinh luong d6ng thM paracetamol va cafein bSng phu'O'ng phap dien di mao quan,
t6i uu hoa bSng phan mem JMP
D a t v a n d e
Viec dinh lugng d i n g thdi paracetamol va cafein bang phu'ang phdp dien di mao quan (CE) hien chua co nhiiu edng trinh nghien cdu. Tgi Viet Nam, CE van la mpt ky thudt phan tich kha mdi, chua d u g c dng dyng rpng rdi. Trong khi do, CE CO nhieu u u diem han so vdi eac phuang phap khae nhu: thdi gian phan tieh n g i n , lugng mau va dung m6i tieu ton it, ban ehdt pha d6ng it dpe hai. Viee m d rpng ung dung va tieu ehuan hoa phuang phap nay rat c6 Igi trong thye t i i n , do do de tdi "Xdy dyng quy trinh djnh lugng d i n g thdi paracetamol va cafein bdng phuang phap dien di mao quan, toi u u hoa bdng phan m i m JMP" la budc dau lam phong phu pham vi ung dyng ciia phuang phdp CE.
N g u y e n l i e u v a p h i p c n g p h a p Nguyen lieu
- Vien nen Y 1 , Y2, Y3 eo e6ng thde:
Paracetamol : 500 mg Cafein : 65 mg Ta d u g c v u a dii : 1 vien
- Paracetamol (dp I m 0,10 %; ham l u g n g 99,86 %) vd cafein (dp am 0,11 %; ham l u g n g 100,01 %) CO n g u i n g i c t u Vien Kiem nghiem thuoe TP. H i Chi l^inh, sulfanilamid (do am 0,08 %; ham lugng 99,9 %) san x u i t bdi Hang Merck.
- Dinatri tetrabcrat logi P.A (Merck), natri dodecylsulfat (SDS) logi P.A. (Merck).
Thiet bj
- May dien di mac quan Agilent CE-7100, d i u d6 PDA, phdn mem Chemstation.
Nguyen Ngoc Yen Nhi, Vinh Dinh
Khoa Dugc - Dai hgc YDugc TP. Hd Chi Minh- Cpt mao quan silicagel nung chay dudng kfnh trong 50 pm, chieu dai t i n g cpng 64,5 cm, chieu dai hipu l y e 56 cm.
P h u c n g phap nghien c u u
D y a vao p h u a n g phap djnh l u g n g hon hpp CO chda paracetamol vd cafein trong che pham bdng ky thugt dien di mao quan mieell didn d6ng ''• ^' •*', xdy d y n g qui trinh dmh l u g n g va t i i u'u hoa dieu kien phan tieh bang p h i n m i m JMP'^^, sau do t i i n hdnh danh gid quy trinh dmh luong theo p h u a n g phdp t h u d n g qui.
Chuan bi dung djch chain ngi (IS)sulfanilamid
Can chinh xac 0,013 g ehuan nfii sulfanilamid cho vao binh djnh m d c 50 ml.
Them mpt ft n u d e cdt k h u khoang, l i e d i u , sieu am cho tan r i i dien n u d c d i n du vach.
Chuin bj dung dfch doi chieu Cdn ehinh xac khoang 0,100 g chuan paracetamol va 0,013 g c h u i n cafein cho vao binh dinh m d c 20 ml, them khoang 14 ml nuoc cat k h u khoang, Idc d i u , sieu am cho tan roi them n u d c d i n v d a du d i n vgch, Idc deu.
L i y ehinh xac 1 ml dung dich tren vd 2,5 ml dung djch chuan npi eho vao binh djnh mdc 10 ml, d i i n du vdi n u d e cat k h u khoang, l i e deu.
Loc dung djch doi chieu thu d u g e qua mang Ipc 0,2 pm r i i sieu dm k h d khi. Dung djch d i i chieu CO nong dp paracetamol chuan 500 pg/ml, cafein chuan 65 pg/ml va c h i t chuan npi sulfanilamid 65 pg/ml.
Chuan bj dung djch thu-
Cdn 20 vien c h i pham, tinh k h i i l u g n g tmng
TAP CHI DlTQC HOC - 02/2013 (S6 442 N A M S
• Nghien CCFU - Ky thuat
binh eua mfit vien. N g h i i n thanh bpt min. Can mot lugng bpt t h u i c t u a n g ung vdi m6t nda k h i i lugng trung binh vien eho vao binh dinh mde 50 ml. Them n u d c c i t khu khoang, sieu dm 15 phut, de nguoi r i i d i i n n u d e cat khu khodng d i n vach, lac d i u . Lpe dung djch qua giay Ipc, bo 20 ml dieh Ipe dau. Lay 1 ml dung djeh nay va 2,5 ml dung djch chuan noi cho vdo binh djnh muc 10 ml, them n u d e c i t k h d khoang d i n vach, lac deu. Loc dung djch t h d thu d u g c qua mdng Ipc 0,2 pm r i i sieu am khir khf. Dung dich t h u co nong dp t u a n g duang vdi dung dich d i i c h i i u .
Ket qua va ban luan Toi u u hod d i i u kien p h d n tich Sau khi khao sat sa bp d i i u kien dien di, tim dwgc d i i u kien phan tich hieu qua la dem borat 25 mM/L CO pH = 10, n i n g dp c h i t dien hoat SDS Id 25 mM/L T i i p tuc t i i uu hoa diiu kien nay bdng p h i n m i m JMP ( b i tri 15 thi nghiem theo m6 hinh Box-Behnken ) vdi ba y i u t i pH (XI), n i n g do dpm borat (X2) vd n i n g do SDS (X3). Ca ba y i u to tren deu anh hudng d i n dp phan giai cua cae pic nen chpn th6ng so dap dng ciia eac thf nghiem la d6 phdn giai Rsl ciia eafein-IS, dp phan giai Rs2 ciia paraeetamol-cafein.
Bang 1 : Kit qua bd tri 15 thi nghiem theo mo hinh Box-Behnken STT
1 2 3 4 5 6 7 8
Ma hoa - 0 -0- -0+
-+0 0-- 0-+
000 000
X1 9,5 9,5 9,5 9,5 10 10 10 10
X2 20 26 25 30 20 20 26 25
X3 25 20 30 25 20 30 25 25
Rsl 9,65 7,61 12,72 9,35 4,35 7,90 7,99 7,91
Rsl 8,00 7,01 3,61 4,85 17,07 13.09 17.15 17.05
STT 9 10 11 12 13 14 15
Ma hoa 000 0+- 0++
+-0 t o - t o * ++0
X1 10 10 10 10,5 10,5 10,5 10,5
X2 25 30 30 20 25 26 30
X3 25 20 30 25 20 30 25
R s l 7,86 4,65 8,38 1,23 3,62 0 2.64
Rs2 17,05 14,06 15,72 26.42 24.74 25.89 26,45
" ^
7^
-^^^^-i—^ P--+-H-
- ^
Hinh tieh (pH,
1: Ket qua tdi wu hda diiu kien phan borat. SDS) bing phan mem JMP
Sau khi phan tfch bdng p h i n m i m , d i i u kien t i i uu hoa cho ba y i u t i phan tfch la: dung djch dien ly nen dem borat 25,7 mM/L; pH 10,1 co chda SDS n i n g do 26 mM/L; nhiet do 25°C;
dien t h i 25 kV; b u d c song phdt hien 214 nm.
Diing d i i u kien nay t i i p tuc kiem tra tinh t u a n g thfch he t h i n g va danh gia phuang phap dinh lugng.
T i n h t u c n g t h i c h he t h o n g
Chuan bi m l u d i i c h i i u va mdu t h u nhu myc
"Phuang phap nghien cuu" M6i dung djch mau chuan va m i u thu d u g c dien di 6 lan trong ciing dieu kien dien di da xac djnh.
Bang 2: Ket qui khio sit tinh twang thich he thdng cua paracetamol va cafein
S/Si, t/tl.
H R, A,
Paracetamol dol chieu TB
5,039 1,238 19,683 17,605 1.745
RSD%
1,72 0,20 1.45 0,77 0,94
Paracetamol thu TB 5,162 1,242 19,067 17.493 1.720
RSD%
1.666 0.15 1.74 0.54 1,821
Cafein dol chieu TB 1,357 1,091 6,733 7,618 1,055
RSD%
1,507 0.14 1.80 1,1 0,994
Cafein thu TB 1,402 1,092 6,633 7,632 1,091
RSD%
0,95 0.21 0.78 1.48 0.20
TAP CHi Dirpc HQC ~ 02/2013 (S6 442 NAM 53)
• Nghien CLFU - Ky thuat
T i n h dac hieu
C h u i n bj mdu doi chiiu va m i u thu nhu myc
"Phuang phap nghien cdu", mau trdng la dung moi
hoa tan mau (nude eat khiF khoang). T i i n hdnh dien di dung dich mau chuan, mdu thu, mau trang va mau thu thdm chuan. Moi mdu bam 1 l i n .
ro)
( C ) ( B ) ( A )
<2)
,^.n
,. A _
I /I
_j\___
Hinh 2; Dien di dd cua (A) miu tring. (B) miu chuin, (C) mau thw vi (D) miu thw thim chuin d diiu kien toi wu hoa. (1) la sulfanilamid, (2) la cafein, (3) la paracetamol
Hinh 5: Pho UV cua paracetamol ehuan T i n h tuyen t i n h
C h u a n bi cae d u n g djch eo nong dp p a r a c e t a m o l va c a f e i n n h u t r o n g b a n g 2 va b a n g 3. D i e n di cac d u n g djeh d i i c h i i u
Hinh 6: Phd UV cOa paracetamol thw t r e n , m i i d u n g djeh t i e m 3 l a n . Khao sat s y phu t h u o c t u y e n t i n h g i i j a n o n g dp hogt c h d t v d i ty s i C o r r A r e a c u a hogt chat va c h u a n n 6 i .
Bang
STT 1 2 3 4 5 6 7
3: Ket qua khao sat tinh tuyen tinh Paracetamol
(pg/ml) 200 400 500 600 800 1000 1250 Phu'O'ng trinh hoi quy; y
R'
Chuan ngi (pg/ml)
85 65 65 65 65 65 65
= 0.0085 X +
= 0.9995
cua paracetamol S/Su, 1,821 3.491 4.526 5.273 7,037 8,667 10,738 0,1675
STT 1 2 3 4 5 6 7
va cafein Cafein (pg/ml) 200 400 500 600 800 1000 1250 PhLPCTng tr
Chuan no (pg/ml)
65 65 65 65 65 65 65 nh hoi quy y =
R'= 0,9994
S/Sis 1,821 3,491 4.526 5.273 7.037 8,667 10,738 0,0186 X
TAP CHi Dirpc HQC - 02/2013 (SO 442 NAM 53)
» Nghien CLPU - Ky thuat
Dp dung
Dp diing cua phuang phap d u g c xac djnh bdng p h u a n g phap thdm ehuan. Them mot
Bang 5: Kit qua khao sit dg dung cua paracetamol vi cafein
lugng chuan paracetamol va cafein t u a n g dng vdi 80%, 100% va 120% hdm lugng mau t h d . M i i mdu tiem 3 ldn.
80%
100%
120%
Luryng tim thay (mg)
454,14 451,20 453,09 500,26 500,59 496,87 668,14 556,92 551,53
Paracetamol Luvng li' thuyet (mg)
251.78 251.78 251.78 251.78 251.78 251.78 251.78 251.78 251.78
Lu^ng them v^o (mg)
199,8 200,3 200,2 249,9 250,3 249,8 299,7 301,5 300,3 Ty le phuc hoi 1^ thuyet {%): 98-102 Ty le phuc hoi trung 1 3D: 0,642; RSD%; 0,64; e
%
phuc hoi 100,57 99.81 100.25 99.72 99,70 99,06 101,21 100,66 99,90
3lnh- 100,10%
(p< 0,05). ± 0,51%
Luong tim thiy (mg) 58,17 58,92 58,52 64,71 65.45 64.29 71.24 71.72 71.06
Cafein Luong ly thuySt (mg)
32.86 32.86 32.86 32.86 32.86 32,86 32,86 32,86 32,86
Luong thim vao (mg)
26,1 26,3 25,8 32,7 32,5 32.6 38.9 39,0 39,3
%
phuc hdi 98,67 99,59 99.76 98,70 100,14 98,36 99,28 99,80 98,47 Ty le phuc hoi ly thuyet (%): 98-102
Ty le phuc hoi trung binh' 99.20%
SD: 0.660; RSD%' 0.67; e (p< 0.05); ± 0.52%
Dp chinh xic vi dg chinh xac lien ngiy Chuan bi dung djeh 6 mau thu nhu myc Phuang phap nghien cuu. Tien hanh dipn di 6
Bang 7: Kit qua khao sit dd chinh xic vi dg chinh xac lien ngay
dung dich mdu t h d d dieu kien da xac djnh, m i l dung djch bam 1 lan. Xac dinh d6 Ipch chuan tuong doi
Ham Itrgng paracetamol (%) Ham Iwgng cafein (%) Ngay 1
1 97,23 2 98,65 3 99,87 4 97,5 5 96,88 6 97,87 TB 98 RSD% 1,12
TB tong the
Ngay 2 98,29 99,68 98,60 98,02 100.62 100,48 99.28 1.14 99,29
Ng^y3 97.82 102.71 102.10 101.24 100.43 99.23 100.59 1.82
Ngay 1 97.71 99.92 98.29 102.34 98,86 100.72 99.64 1.72
Ngay 2 98.81 97.58 99.80 99.61 96,28 97.53 98.27 1.40 98,36
Ngay 3 99,26 97,66 95,23 95.66 97.68 97.50 97.16 1,53
RSD% tong the 1,72
Ket luan
Phuang phdp djnh lugng d i n g thdi paracetamol va cafein bdng ky thuat dien di mao quan mieell dien dong co dp dung la 100,10 % va 99,20 %, RSD % cua dp ehinh xae lien ngay la 1,72 vd 1,8, t u a n g ung vdi paracetamol va cafein, dgt tieu ehuan chap nhgn v i dp dCing (98-102 %) vd d6 chinh xde (RSD % < 2 % ) . Khoang tuyen tfnh cua paracetamol la 200-1250 pg/ml vdi phuang trinh hoi quy y = 0,0085 x +
TAP CHI DlTQC HQC - 02/2013 (SO 442 NAM 53)
0,1675, R^ = 0,9995; khoang t u y i n tfnh cua cafein la 26-162,5 pg/ml vdi phuang trinh h i i quy y = 0,0186 x, R^ = 0,9994. Ddy id phuang phdp CO dp nhgy cao, co the ap dung d i djnh iugng d i n g thdi paracetamol va cafein trong cac c h i pham tren thi trudng.
S u m m a r y
Paracetamol and caffeine in some pharmaceuticals on the market were analysed
m Nghien C I J U - Ky thuat
by micellar electrokinetic chromatography (MEKC). The electrophoresis conditions were investigated optimizated by JMP software for the background electrolyte cell: pH 10.1;
disodiumtetraborat buffer - 25.7 mM; SDS - 26.0 mM; voltage - 25 kV; temperature - 25°C;
absorption wavelength 214 nm; sample injection 3s x 50 mbar. The method was validated proving desirable specificity, linearity, precision (RSD < 2%) and accuracy (98-102%).
Key words: Micellar electrokinetic chromatography, survey, optimization, JMP software..
T a i l i e u t i i a m k h a o 1. David Heiger (2000), High performance capilary electmphoresis. pp. 25, 37-43, 49.
2. i<3Jkieep Delvadiya et a/ (2011), "Spectrophotometric simultaneous analysis of paracetamol, propyphenazone and caffeine in tablet dosage forms', Internatbnal journal of pharmacy and pharmaceutical sciences. 3(3), pp. 170-173.
3. SAS Institute Inc. (2005), Jfi4P User Guide. CD- ROM JMP version 4.0.
4. S. Bookerd et al (1995), "Separation and simultaneous determination of the components in an analgesic tablet fonnulation by micellar electrokinetic chromatography", Journal of Chromatography A. 695, pp. 98-102.
r
Nghien cii'u xay dyng cong tliirc bao clie pellet omeprazol bao tan o" ruot
f ) a t v a n d e
Bao c h i pellet bao tan d ru6t eho d u g c chat omeprazol rat kho khdn do omeprazol kem ben vdi nhiet, am^^\ dnh sdng'^', pH acid'^' va chat tao mdng bao Eudragit LIOO'^ V i vgy, de xay dung d u g c cong thde bao c h i pellet omeprazol on dinh, dam bao c h i t l u g n g pellet, ehung toi t i i n hanh d i tdi "Nghien cuti xay dyng cdng thiTc bio che pellet omeprazol bao tan &
rugt. vdi myc tieu la ddnh gia d u g c cdc yeu t i ve e6ng thuc len chat lugng pellet omeprazol.
N g u y e n i i e u v a t i i i d t bj n g i i i e n C I J U Nguyen lieu
Omeprazol magnesi (omeprazol), hydroxypropylmethyl cellulose (HPIVIC) E l 5, HPfVlC E6, polyvinyl pyrolidon K30 (PVP), natri laurylsulfat, dinatri hydrophosphat (Na2HP04), Aeryl-EZE (Eudragit L100), dibutyl phtalat (DBP) va talc: dgt tieu chuan d u g e dung. Cdc hoa c h i t khac: dgt tieu chuan hoa chat phan tieh. Vien nang Lomac 20mg co so 16: X11099, han s u dung: 01/08/2014.
Thiet bj nghien c d u
Hp thong bao t i n g soi Diosna, mdy t h u dp
10
Nguyen Ngoc Chien, Tr|nh Huy Cong
Truang Dgi hgc Dugc Hd Ngi hoa tan ERWEKA DT60, may quang pho UV-Vis Hitachi U-1900, tu say IVlemmert, can phan tich Sartorius, he t h i n g sde ky long hidu nang cao SHIMADZU IVIQ 022, kinh h i i n vi dien t d quet (SEIVI) HITACHI S-4800.Phuxmg phap nghien c u u
Phuvng phap danh gia tuvng ky daw chat vdi dung moi vi polymer
Dung mdi bdi va bao mang each ly Theo doi mdu sdc cCia h i n dich chda 5 % d u g c chat trong cae dung m6i, s u dung ong nghipm thuy tinh 20 ml trong s u i t , nut kin.
Dung djch polymer: T u a n g t y phuang phap tren, s d dyng hon djeh chua 5 % d u g c c h i t v a 5 % polymer dung bao c h i pellet trong dung djch Na2HP04 1 %.
Phuvng phap bao che pellet om^ran^
nhan
Tien hdnh boi len pellet tra (chda 40 % lactose^ 60 % Avicel PH 101, kieh t h u d c 0,8-1,2 mm) h i n dieh d u g e chat, s u dyng t h i i t bj tang s6i vdi eac th6ng so: dp s u i t khi phun = 1,8-2,0 bar; ap luc gio = 80 %; nhi^t dp khf vao = 50°C, t i c dp phun djch = 6,6 ml/phiit, d u d n g kinh v6i
