Nghien CLPU - Ky thuat
iTng dung phan mem Matlab va JMP trong
r
nghien ciru toi tfu hoa phan u'ng khfr hoa phthaloyl amlodipin thanh amlodipin base
Dang Minh Quan, Tran Thanh Dao, Le Minh Tri
Bp mon Hoa Dugc, Khoa Duac, DHYD TP. HCMDat van de
Amlodipin la mot thuic tim mach thupe nhom Chen kenh caici, eo tae dung t i t trong d i i u tri tang huyit ap va d i i u trj du phong a nguo'i benh dau thit ngue t h i i n dinh. Nguyen lieu cho viec bao c h i cac san p h i m eo hoat c h i t amlodipin cua nuac ta hien nay phin lo'n nhap tCp nuac ngoai, vi t h i d i chu dpng dugc nguin nguyen lieu c i n nghien cu'u eae qui trinh t i n g hgp amlodipin n h i m tao ra amlodipin dat tieu chuin dugc dung va co hieu suit cao. De t i n g hgp amlodipin phai trai qua nhiiu giai doan khae nhau, trong pham vi d i tai nay chung t6i se Cpng dung phin m i m Matlab va JMP d i nghien CCPU t i i uu hoa giai doan khCp hoa phthaloyl amlodipin thanh amlodipin.
Nguyen lieu, thi§t bj va phu'ang phap
Nguyen lieu: phthaloyl amlodipin, methylamin (dd 40% trong nuo'c), amlodipin chuin, toluen, ethanol, ban s i c ky lo'p mong trang s i n Kieselgel 60F 254: dat tieu chuin phan tich.
Acetonitril, amoni acetat, methanol, triethylamin (Merck) loai dung cho HPLC.
Thiit bi nghien CCPU: bp phan Cpng 4 c h i Eyela/Tokyo Rikakikai, buing soi UV Vilber Lourmat, may anh Canon A580, may s i c ky long hieu nang cao Waters 2695, phin m i m Matlab R2009a va JMP 4.0. May do NMR Bruker 500 MHz, may do IR FTIR-8210 PC, may k h i i p h i Quattro Micro API.
Phircng phap nghien ciru
Khao sat va giai han cac yeu to anh hwang den hieu suat
Tiin hanh phan Cpng a eae diiu kien tham do'^';
Nhiet dp; 45-90 °C, tho'i gian: 3-20 gio', ti le moi phthaloyl amlodipin : methylamin la 1:40-1:85.
Danh gia hieu suit phan Cpng bing phuang phap ban djnh lugng TLC dua tren s u thay d i i dien tfeh v i t s i c ky. M i u dugc dua len ban mong bing i n g mao quan ehfnh xac 5 pl, c h i m 4 lin va triin khai s i c ky vo'i he dung moi:
Ethylacetat-methanol-amoniae 25% (20 : 4 : 1).
Sau do, dien tich v i t s i c ky se duge tinh toan bing each SLP dung p h i n m i m Matlab lap trinh chuang trinh XLP ly anh'"'.
Nguyen ly hoat d6ng cua chuang trinh XCP ly anh: Hinh anh duge e i u tao tCp r i t nhiiu cac d i i m nho gpi la d i i m anh hay pixel. Anh mau (RGB) dugc chuyen doi thanh anh xam (moi diem anh duge loai bo cac th6ng tin v i mau, giu lai nhu'ng th6ng tin v i dp sang) va mang gia trj trong khoang 0-1.
Anh xam (grayscale) duge t i i p tue chuyen d i i thanh anh nhj phan (tring den) vai he s i mCpc chuyen doi X thfch hgp: cac diim anh mang gia trj lo'n han he s i X se nhan gia trj 1 va tra thanh mau tring; cac d i i m anh mang gia tri nho han he s i X se nhan gia trj 0 va tro' thanh mau den.
TCP anh nhj phan, hinh anh dugc xem nhu la m6t ma tran vo'i nhiiu hang va c6t, SLP dung ham nghjeh dao de chuyen gia trj 0 thanh 1 va 1 thanh 0, theo do tfnh t i n g ma tran thi se thu duge t i n g gia trj cac d i i m anh mau den, chinh la dien tich pixel cua vit sic ky khao sat
Bo tri thi nghiem, xac djnh thong so toi wu bang phan mem JMP.
Vo'i mCpc gio'i han cua cac y i u t i anh hugng den hieu s u i t da dugc xac djnh, SLP dung phin mem JMP d i b i tri thf nghiem b i n g phugng phap dap Cpng b i mat theo mo hinh Box- Behnken'^'. Ma tran quy hoach thuc nghiem thu dugc g i m 30 phan Cpng (eae phan Cpng duge lap lai them 1 l i n 6e tang dp tin cay eua s i lieu).
44
TAP CHi DUQC HQC - 11/2010 (SO 415 NAM 50)
Nghien CLPU - Ky thuat
Thong s i dap Cpng cua ma trSn quy hoach thuc nghiem la hi^u suit se dugc danh gia bing phuang phap djnh lugng HPLC v6'i diiu kien sic ky;
C6t: Luna CI 8(2) 100A, 250 x 4,60 mm, kfeh thuo'c hat 5 pm.
Pha dpng: methanol-aeetonitril-dd muii amonium acetat 40 mM (38:24:38), trong do c6 ehCra 0,02% triethylamin, pH=7,1.
Tic dp dong: 1,2 ml/phut.
Thi tfeh tiem miu: 25 pl.
Diu do: day died quang, quan sat 6' buo'c s6ng 239 nm.
Kit qua thu dugc tCp 30 thf nghiem se duge XLP ly bing phin mim JMP: xac djnh mil tugng quan giu'a eae yiu t i nhiet dp, tho'i gian, ti le moi dii vo'i hieu suit eua phan Cpng; xay dung
phugng trinh be mat dap Cpng va du doan diiu ki$n phan Cpng cho hi$u suit t i i uu.
Kiim nghiem san phSm
Amlodipin diiu che dugc xac djnh cau tmc bing phuang phap do quang phi hip thu hing ngoai, H- NMR va MS. De dinh lugng, nghien
CCPUnay dung phuang phap HPLC-UV a buo'c song 239 nm'^l
K§t qua nghien ciru
Lap trinh chiFO'ng trinh xu> ly anh tinh toan dien tich vit sic Icy
Viit chuang trinh bing ng6n ngCp Matlab, v&l su h i trg eua hop e6ng cu toolbox processing image cac thuat toan nhim xu' ly hinh anh se duge thuc thi mot each d i dang hgn. Cac ma nguin eua chugng trinh se duge thi hanh ngay tren phin mim ma khong cin phai bien djch qua eae tap tin thi hanh.
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nong do 0,5% Dien tich = 836
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Hinh 1: Chwang trlnh xCr ly anh f Cae anh ehup phai ding nhit v i dp sang, dp
phan giai, cu ly chup...do do cin c i djnh may anh g eung chi dp va khoang each. Chpn chi dp Manual trong may anh khi ehup vo'i th6ng s i cuang dp sang - 1 — , d6 phan giai 2 Megapixel.
Xay dipng dirvng hdi quy tuyin tinh dien tich vit cua miu chuin amlodipin theo cac ning dp 0,5%, 1%, 1,5%, 2%, 2,5%, 3%.
Bang 1: Kit qua khao sat dien tich vit theo ndng dp
tinh toin dien tich vit sic ky
a dp tin cay 95%. Nhu vay, e6 the SLP dung phuang trinh hoi qui tuyen tfnh nay de xac djnh ning dp amlodipin cua miu thCp dua tren dien tfch vit.
Giai han khoang khao sat cua cac yiu td anh hu'O'ng
Yeu to nhiet dp
Bang 2: Kit qui khio sit yiu td nhiet do
Nong dp
amlodipin 0,5% 1% 1,5% 2% 2,5% 3%
Dien tfch vet
(pixel) 836 1135 1401 1621 1830 2127 Phuang trinh hii qui tuyin tfnh dien tfeh vit theo ning dp eua miu chuin amlodipin co dang:. Y = 0,002X - 1,2193 vai he s i R^= 0,9964
TfC) 45 60 70 70 70 70 80 90
t (gia) 16 16 16 12 8 4 4 4
Ti le moi 55 55 55 55 55 55 55 55
Dipn tieh vet (pixel) 703
846 1647 1694 1358 1346 1541 1644
H(%) 3 7,7 32,8 35,3 24,4 24 30,4 33,7
TAP CHi DUQC HQC - 11/2010 (SO 415 NAM 50) 45
• Nghien CLPU - Ky thuat
Khi tang nhiet dp, hieu suit tSng va giam thb'i gian phan Cpng. Do vay, gi6'i han mCPC biin thien y i u t i nhi$t dp duge xac djnh trong khoang 70-90°C.
Yeu to thai gian
Bang 3: Kit qua khao sat yiu td thai gian
TCC)t(gia)
80 3 80 4 80 5 80 6 80 7
Ti le moi 55 55 55 55 55
Dien tieh vet (pixel) 1154 1541 1454 1452 1426
H(%) 17,7 30,4 27,5 27,5 26,6
thd-i gian dugc xac djnh trong khoang 3-5 gio'.
Yeu to tile moi
Bang 4: Kit qua khao sit yiu td ti le moi T CC) t (gia) Ti le moi Di^n tieh vet (pixel) H (%)
80 4 40 1618 32,9
80 55 1541 30,4
80 70 1418 26,4
80 85 1455 27,9
Khi tang tho'i gian thi hieu suat tang, nhung d i n mot thai d i i m thi hieu suit kh6ng tang nCpa.
Do vay, gio'i han khoang khao sat cua y i u t i
Thong s6 t i i u'u va tu'ang quan giifa cac y i u t i anh hu'O'ng d i i vai hieu s u i t
Khi ti le moi tang thi hieu s u i t giam. Tuy nhien, methyiamin/nuo'c eon dong vai tro la dung m6i phan Cpng nen lugng eho vao phai du 6k hoa tan phthaloyl amlodipin, do vay gio'i han khoang khao sat y i u t i ti le moi eua phthaloyl amlodipin : methylamine la 1:40-1:70.
15 20 H i a j s u d RMSE=4,
25 30 35 « 45 t Predicted P<,0001 RSq=0,83
,6601
Term Intercept Nhetd3(70,a5&RS Thd gian(3,5)&RS Tiiemol(«l,7D)8RS Ntiet d:X70,9C(*Ttioi gian(3,5) Ntiet *(70,9C}*Ti lemd(4),70) Thdgian(3,5)'Tiiemcl(«),70) N r t d dD(70,9CCNilietdo(70,90) T I r i gian{35)'Tlxi gian(3,5) Ti lemc((40,70)Tl lemol(40,70)
2,7122917 1,4910417
S H E r r a 1,898«3 1,16253 1,16253 1,15253 1,644066 1,64406 1,544066 1,711198 1,711198 1.711198
t R a l c 1567 8,47 1,86 -0,93 0,38 2,53 -3,37 -0,28 1,59 0,87
Prolyl
<.0001
<.O0Ol 0,0783 0,3610 0,7112 0,0199 0,0031 07791 0,1286 0,3939
Hinh 2: Biiu dd thi hien twang quan Su tuang quan tuyin tinh giO'a cac y i u t i d i i VO'I hieu suit la eo y nghTa vo'i gia tri R^ = 0,83 a dp tin cay 95%. Phuang trinh dap Cpng b i mat:
Y = 29,75 + 9,84Xi + 4,16X1X3 - 5,54X2X3 X , ; nhiet dp phan wng.
X3: tile moi cic chit phan wng.
X2: thai gian phan wng.
Y: hieu suit phin wng.
Qua phuang trinh b i mat dap Cpng tren, ta
tuyin tinh cua cic yiu td v&i hieu suit
thiy cac tham s i v i nhiet dp, thtji gian, ti le moi phan Cpng d i u co anh huang d i n hieu suit phan Cpng. Hieu s u i t phan Cpng dat gia tri t i i da la Y= 49,14% VO'I Xi=90 °C, X2=3 gio' va X3=70.
Bing c6ng cu Prediction Profiler trong JMP CO t h i tim duge eae gia trj khac cua y i u t i anh huang ma v i n cho hieu s u i t eao: Y= 48,55%
vai Xi=90 °C, X2= 5 gia, X3= 40. Thong s i nay CO Igi d i i m v i kinh t i han (ti le moi phan u'ng ft han nen SLP dung ft hoa c h i t methylamin han).
1 P r e d i c t i o n P r o f i l e r |
56.65 _
i
15,102 _
• -.: ;
1
••' i • - v . f * '
-••'•• •
i
NiTiet do(70.90) T h d gian(3,5) Tileird(40,70)
Hinh 3: Cdng cu Prediction profiler xic djnh hieu suit
46 TAP CHI DUQC HQC - 11/2010 (SO 415 NAM 50)
N g h i e n CLPU - K y t h u a t
Tiin hanh phan Cpng 3 lin a dieu kien du doan, kit qua hi§u suit thu dugc la: 47,7%.
Tiin hanh tinh chi amlodipin base, kit qua hi§u suit sau tinh che: 31,4%.
Cac kit qua kiim nghi§m, djnh lugng va xac djnh ciu true eua amlodipin base diu dat yeu eiu.
Kdt luan
Da lap trinh dugc chugng trinh XLP ly anh de tinh toan dien tich vit sic ky. Oa xac djnh dugc cac th6ng s i tii uu cua phan Cpng khCp hoa phthaloyl amlodipin thanh amlodipin: nhiet dp:
90 °C, tho'i gian: 5 gio', ti le moi cua phthaloyl amlodipin : methylamine la 1:40. Da tinh ehi dugc amlodipin sau phan Cpng, vo'i hieu suit sau tinh chi: 31,4%.
Summary
An image processing program was made by Matlab software to measure the area of TLC spot
It was helpful in quantitative analysis by TLC.
With the support of JMP software, optimal parameters for the reaction of making amiodipine base from phthaloyl amiodipine were determined:
temperature 90 °C, time: 5 hours, ratio of phthaloyl amiodipine and hydrous methylamine: 1:40.
Keywords: phthaloyl amiodipine, amiodipine
base, Matlab software, JMP software.
Tai lieu tham khao
1. George Lunn and Norman R. SchmLJff, Methods for Pharmaceutical analysis, John Wiley & Sons, Inc, NewYork, (1997), pp 87-88.
2. Lemmens et al., Amiodipine free base. United States Patent 7335380 (2008).
3. John Sail, Lee Creighton, Ann Lehman, JMP DOE Guide, In : JMP Start Statistics, SAS Institue Inc., Cary, NC, USA,(2007), pp 69-83.
4. Processing image toolbox User's guide, MathWorks Inc., Natick, MA, USA 1993-1997.
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Danh gia sinh kha dung cua thuoc bot
m r
cefuroxim pha hon dich uong
Dat van de
Cefuroxim duge SLP dung r6ng rai trong diiu trj nhCpng benh nhiim trung do vi khuan gram duang va vi khuin gram am ke ea nhu'ng chung vi khuin san sinh betalactamase gay ra.
Cefuroxim thuo'ng duge ehi djnh SLP dung trong nhiim khuan h6 hip duo'i/ tren; nhiim khuin nieu, sinh due; nhiim khuan da va m6 mim '^l
Bpt pha hin djch la mot dang thuic co nhiiu uu diim. Vi la dang bao chi rin nen de phan liiu chinh xac, d i van chuyen, bao quan, khie phuc dugc tfnh kem bin cua mot s i dugc chit niu dugc diiu chi duo'i dang long, dae biet la cac khang sinh. Thuie bpt pha hon dich uing thfch hgp eho tre em va nguo'i lo'n tuii, nhCpng dii tugng kho uing thuic dang rin. Tuy nhien, cefuroxim la mot chit co vj rit ding va keo dai, rit kho uing duo'i dang long khi duge phan tan thanh hin djch. Vi vay, trong qua trinh dieu ehi, TAP CHi DUQC HQC - 11/2010 (SO 415 NAM 50)
Dirong Thi Kim TrinhS Trinh Thj Thu Loan^
Cong ty CO phan Duac Pham Imexpharm,
^Khoa Dugc, DH Y Dugc TP. HCM
nhu'ng ky thuat dac biet duge ap dung 6e ehe vj ding rit kho chju eua cefuroxim nhung van phai dam bao duge yeu eiu v i mat kiem nghiem va c6 sinh kha dung eao de dam bao hieu qua trj lieu eua thuie. Sinh kha dung eua thuie la eg sa quan trpng giup cho viec lua chpn thay t h i thuic va trong nhieu truo'ng hgp giup giam chi phf diiu trj cho benh nhan.
Mue tieu cua d i tai la nghien
CCPUdanh gia sinh kha dung cua thuic b6t cefuroxim so sanh vai thuic dii chiiu zinnat.
D6i tu'O'ng va Phu'O'ng phap nghien
CU'U
Nguyen lieu va thiit bi
Cefuroxim natri va cefalexin chuin (Vien kiim nghiem thuie TP.HCM), acetonitril dung eho HPLC (DCPC), methanol dung cho HPLC
(DCPC),
natri dihydrophosphat, dinatri hydrophosphat tinh khiit. May HPLC (Elite
47
