Y Hpc Vl|T NAM THANG 8 - s 6 1/2013
HIEU LUIC HOA PHimHG PHAP DjNH LUfOTNG CEFACLOR TRONG HUYET TlTONG NGU6l VOTI SAC KY LONG HIEU NANG CAO
Tran Xuan T r i * , Biii Tung H i S p * * , Nguyen Minh C h i n h * * * sinh nhdm cephalosporin the he thff nhat.
Cefaclor cd hieu qua trong dieu tri mot so benh nhiem trimg dffdng hd hap va tiet nieu, md mem, nhiem trung cdng dong mac phai khac'^'^^
Ngoai ra, cefaclor dffdc dung nap tdt. Nhffng nghien cffu trong dng nghiem cho thay thudc cd hoat tfnh tren vi khuan E. coll, K pneumoniae, P.mlmbllis, H. influenzae, Gonococci, va tu cau khuan ^^•^\
Tru'dc day da cd vai phffdng phap da dffdc nghien ciTu de xac dinh Cefaclor tren ngddP'^'^^K Trong sd dd, mdt phffdng phap phat hien vi sinh hpc da dddc thffc hien, trong khi nhutig nghien cffu khac, phffdng phap sac ky Idng hieu nang cao (HPLC) diing de xac djnh mdt hoac mdt sd cephalosporin cung da dffdc thffc hien.^^°''^'^^^
Phffdng phap xet nghiem vi sinh djnh Iffdng khang sinh theo kha nang khang vi sinh vat. Tuy nhien, cac phffdng phap dd khdng the xem la chuyen biet nhff phffdng phap dinh Iffdng bang HPLC. Ke tff khi phffdng phap ciia Signs va cdng sy^'"^ dddc phat trie'n de xac dinh mdt vai cephalosporin, viec sff dung gradient pha dpng de tach cac cephalosporin khac nhau la rat can thiet. Trong nhffng nghien cffu khac ^^^•^^\
cefaclor dffdc xac dinh bang tia cffc ti'm hoac bang dau dd khdi pho. Trong mdt nghien ciiti, dd nhay ciia phffdng phap nay khdng dii tdt de theo dudi thdi gian chay nhieu hdn 4 gid*^^^', trong khl dd, mac dii cac phu'dng phap khac''^' cd do nhay rat tdt, nhffng ddi hdi phai sff dyng cac thiet bi dat tien khdng cd s i n d hau het cac cac phdng thi nghiem.
II. 0 6 1 TUpNG VA PHUONG PHAP NGHllN COU 2.1 Thudc thff va d u n g m d i :
Cefaclor (CEF) (Hinh 1) do cdng ty Merck Mexico S.A. (Mexico City) cung cap trong khi chuan npi 2-acetamidophenol do cdng ty Sigma Chemical Co. (St. Louis, MO) cung cap. Methanol va acetonitrile chaat IddTng sou duing (UV) do cdng ty Merck (Darmstadt, Germany) cung caap.
T 6 M TAT
Mot phu'Ong phap djnh lu'dng nhanh va ddn gian cefaclor trong cac mau huyet tu'dng ngu'di bSng sac ky long hieu nang cao (HPLC), da du'dc hieu lu'c hda.
Phifdng phap nay bao gdm sff khff protein vdi axit percloric ddng thdi la phu'dng phap chiet xusit. Chiet xuat huyet tu'dng dUdc phan tfch tren mot cot pha dao v6i pha dpng la hon hpp acetonitnl va dung djch natri dihydrogen phosphate 0,01 M, dieu chinh pH den 6,5 b§ng kali hydroxyd (7:93, v/v), va ph^t hien hap thu d dp dai scing 260 nm. Thdi gian fu\j ciia cefaclor va chat diuan noi lan lu'dt la 5,2 va 7,2 phiit. Phu'dng phap dinh lu'dng nay tuyen tfnh trong khoang 0,5-30 nl/ml va gidi han phat hien la 100 ng/ml. Phu'dng phap nay hffu ich va phii hop de xac d|nh cefaclor sau khi uong d ngu'di t'nh nguyen khde manh, va cd the du'dc sff dung cho nghien cffu smh kha dung va tu'dng du'dng sinh hpc ciia thuoc.
SUMMARY
VALIDATION OF CEFACLOR IN HUMAN PLASMA SAMPLES BY HPLC
A rapid and simple method for determination of cefador in human plasma samples, by high- performance liquid chromatography (HPLC), was developed. This method includes a deproteinization with perchloric acid as extraction procedure Plasma extracts were analyzed on a reversed phase column eluted with a mixture of acetonitrile and 0.01 M sodium dihydrogen phosphate solution, adjusted to pH 6.5 with potassium hydroxide (7:93, v/v), and detected by absorbance at 260 nm. Retention times for cefador and the internal standard were 5.2 and 7.2 min, respectively. The method was linear in the range of 0.5-30 nl/ml and the detection limit of the method [ was lOOng/ml. This method was useful and suitable ' for determination of cefador after oral administration , in healthy volunteers, and could be used for ' bioavailability and bioequivalence studies ofthe drug.
Keywords: Cefador; Pharmacokinetics; HPLC;
Human plasma
I.
DAT VANoi
Cefaclor la cephalosporin the he thff hai, nhffng nd cung the hien cac dac tfnh cua khang
* Tnrdng Dai hpc Ykhoa Pham Ngpc Thach, Tp HCM;
**Benh viin Cap cuV Truhg vddng Tp HCM;
***'HOC vien Quan Y-Bp Qudc phong Phan bien khoa hpc: PGS.TS. Tran Thanh Nhan
Y HQC VigT NAM T H A N G 8 • s 6 1/2013
Nffdc dffdc khff ion qua he thdng cpt Catlonit va Anionit (Continental Water Systems, El Paso, IX).
NhOng thudc thu' vS dung mdi kh^c dfeu cd chat lu-png dCing trong phSn tich.
2.2 Dung djch me:
Dung dich me cCia Cefaclor dffdc chu^n bj trffdc d ndng dd Img/lml trong nffdc vd dung djch chu^n ndi 2-acetamidophenol dirdc chu^n bj trong methanol. Dd 6n djnh duac kl^m tra trong thdi gian td'i thi^u Id ba th^ng 6 nhi6t dd 4''C.
Dung djch me sau dd dddc pha loSng vdi nffdc 34 dddc nhffng dung djch vdi ndng dd sff dung khdc nhau tff 0.5 d^n 30 Ml/ml Dung djch chu^n ndi 2- acetamidophenol dffpc chulin bj vdi ndng dd co djnh 25 /.il/ml trong nffdc. Pha cdc dung djch Natrium acetat 0.1 M (pH 5.2) va dung djch Natrium dihydrogen phosphat 0.01 M chinh tdi pH 6.5 vdi Kalium hydroxid trong nffdc khff ion.
2.3 H$ thofng sac ky long Achiral Chiral:
He thong sac ky bao gBm mpt bdm mau phan tich tff ddng Wisp 717 plus (Waters, USA), mdt bdm pha dpng Model 515 (Waters Assoc, Mliford, MA),
May dpc Model 2487 (Waters), thffc hien d dp dai sdng 260 nm va noi vdi may ghi Millenium (Waters).
Chat chiet dffdc btfm qua Cot Cs 150 x 4.6 mm ID reverse-phase Xterra'* (5 pm) (Waters)
Pha ddng Id mOt hon hp-p gdm acetonitril, dung djch 0.01 M Natrium dihydrogen phospha^
dieu chinh pH 6.5 vdi kalium hydroxid (7:93, v/v) du'pc chfnh IU'U lu-png 1 ml/phut, d" nhi^t C(f phdng.
2.4 Phffdng phap chiet xuat:
^ ^ CH — C O —HN ^ ^ _ ^
COOH
Hinh 1. Cdng thffc hda hpc cua cefaclor (1) va chuaSn noai (2), 2-acetamidophenol.
Them 100^l dung dich chu^n ndi 2- acetamidophenol (25 pg) vao 0.5 ml huyd't tddng. Sau khi them 600 \i\ hoan hdip methanol- natrium acetat O.IM pH 5.2 (70:30, v/v) va 20 ^1 perchloric acid, mau thff dffdc vortex trong 1 phut vdi toe dd tdi da va c^p ddng vdi nhiet dd - 20''c trong 10 phut. Quay ly tam 5 phut vdl tdc dd 4500 vdng/ phut de tach hai pha. Tach pha hffu cd (Idp 6 tren), bdm trffc tiep lOOjil vdo he thdng sac ky Idng.
2.5. Hi$u li/c hda phUdng phap:
Tinh ddc hidu dupc tham djnh qua bdm huy^t tuong trang (khdng th§m CEF) sau khi chi^t.
Nhffng mlu huyd't tddng diJpc them vdi dung dich CEF de cd dupc ndng dd 0.5-30 ng/ml. chuan npi sff dyng d nong dp co djnh 25 ng/ml.
Tuyg'n tfnh ddpc xdc djnh tren 3 dddng chuan vdi CEF. Phddng trinh hoi quy da dffdc trac nghidm ve tinh tffpng thfch vdi CEF, dffa tren nhffng t? sd (dien tich cua pic cila CEF
trdn didn tfch pic cCia chSft chuan ndi) vdi nhiJng nong dp CEF khac nhau.
Dp chinh xac cila phffpng phap dffpc danh gia qua hieu Iffc hda trong ngay va lien ngdy.
• DO ldp l9i trong ngdy; phdn tich 5 mlu thir a ndng dd th^p 0 5 ng/ml CEF vd nong dO cao 30 ng/ml CEF de tinh h$ s6 bifen ddi trong ngdy CV (Coefficient of variation intraday),
• DO l$p Igi lien ngdy du'p'c khao sdt qua 5 m^u thu- d ndng dO th^p 0.5 [iglm\ CEF vd ndng dp cao 30 ng/ml CEF thyc hi$n trong 3 ngdy lidn tifep.
• DO dung: (Accuracy) du-p-c phdn tich qua nh&ng mdu thif vd-i lupng CEF ly thuyet vdi cCing nong dp diing cho du'dng chuan nhu'ng si>
dyng nh&ng dung djch me khdc nhau.
• Gidi han dinh lu'p'ng: (LOQ; Limit of
quantification) du-pc danh gid qua phdn tich
nhu'ng mdu huy4t tu-ang cd ndng dd CEF them
vdo giam d^n. Gid'i hgn djnh lu-ang dupc xie
djnh vdl nong dd gidi han thap nhdt xdc djnh win
h$ so bien ddi CV < 20%.
Y Hpc VI|T NAM THANG 8 - SO 1/2013
. Hi#u xuat chi^t: au-pc x4c dinh qua so vS n6ng it> cao (30 (ig/ml) v*i dijn tich pic tu'ong sinh di$n tich pic cua CEF trSn 5 miu thu- huyet ling sau l<hi bom tru'c tilp miu dung dich CEF tifong CO nong d6 CEF thSm vSo thap (0.5 ^g/ml) trang nu-clc,
III. KET QuA VA BAN LUijiN 3.1 sic ky dd:
Sic ky do c6 diioo sau chl4t xuit huyit tuong tring v4 sau chiit xuat mau huyit tu'ong c6 CEF auvo trinh bay trong hinh 2. Thffl gian lull cira CEF va chat chuan noi Bn lu'dt la 5.2 va 7.2 phut
I '
phut phut Hinh 2/ Sac ky do cd dffdc vdi mau huyet tffdng trang (A) va mau huyet tu'dng cd 25ng/m! CEF
V 25ng/ml chat chuan ndi (B) trong dd: 1, CEF; 2, chuan ndi, 2-acetamidophenol
3.2 Hieu liTc hoa:
Kiem tra nhutig chat giao thoa cd cung thdi gian Iffu vdi CEF va chat chuan not 2 acetamidophenol: khdng cd chat giao thoa hien dien.
Dudng chuan cua CEF tuyen tfnh trong khoang khao sat tff 0.5-30 ng/ml.
He sd tffdng quan R, dp ddc, intercepts cua CEF dffdc trinh bay trong bang 1
Bang 1: Thdng sd cila phapng trinh hoi quy tuyd'n tinh xac dinh tren nhffng mau thff huyet tupng cd CEF ( n=3)
C E F
H^ sd'tifdng quan 0,997
D p d o c 5,91 Gidi han dinh lucjng la 0.1 ng/ml (CV= 5%)
Do lap Igi trong ngay va lien ngay duoc trinh bay trong being 2 Binq 2:06 l | p lai iiSn nqSy vS trong nqSy cua phu'Cnq phSp dinh luonc
CEF
Nonq dQ iiq/ml 0.5
•30
Trong ngay (%) (n = 5) 16,2 10.3
Intercepts 0,60
CEF
Lien nqay(%) (3 nqav) 18,9 16,6
Y Hpc VigT HAM THANG 8 - SO 1/2013 DO dung tim th^y Id 5 , 1 % vdi CEF
Hi$u xu^t chi^t CEF ( ± CV) tlnh du-pc d ndng dp 0.5 ng/ml Id 65.7% (±15,6) vd d ndng dO 30 ng/ml Id 58.8% (±16.2),
V. KET LUAN
Phu'O'ng phdp djnh lu'png CEF trong huy^t tu'ong ngu'di vdi ky thu^t HPLC dd du-pc hiOu lyc hda se giiip cho vi$c nghi6n ciru trong khdo sdt dupc dpng hpc ciia CEF trftn ngu'di, vd khao sdt kha dyng smh hpc (Biodisponibilitd), tlnh sinh tu'O'ng ddng {Bioequivalence) cCia cdc dgng bdo ch^ generic
TAI UEU THAM K H A O
1. Sourgens, H.; Derendorf, H.; Schifferer, H.
Pharmacokinetic profile of cefaclor. Int. J. Clin.
Pharmacol. Ther. 1997, 35(9), 374-380.
2 Brumntt, W.; Hamilton-Miller, J.M. Cefaclor into the millennium.!. Chemother. 1999, 11 (3), 163-178.
Bach, VT.; Khurana, M.M.; Thadepalli, H. In vitro activity of cefaclor against aerobic and anaerobic bacteria. Antimicrob. Agents Chemother. 1978, IS ( 2 J, 210-213.
Watanakunakorn, C ; Glotzbecker, C.
Comparative susceptibility of Haemophilus species to cefaclor, cefamandole, and five other cephalosporins and ampicillin, chloramphenicol, and tetracycline. Antimicrob. Anents Chemother.
1979, 15(6), 836-838.
BenneR, J.V; Brodie, J.L.; Benner, £.3.;
Kirby, W.M.M. Simplified accurate method for antibiotic assay of clinical specimens. Appt, Microbiol. 1966, H 170-178
Bloch, R.; Szwed, 3.3.; Sloan, R.S.; Luft, F.
Pharmacokinetics of cefaclor in normal subjeds and patients with chronic renal failure. Antimicrob.
Agents Chemother. 1977, 12(6), 730-732.
Spyker, D.A.; Thomas, B.L.; Sande, M.A.;
Bolton, W.K. Pharmacokinetics of cefaclor aixf cephalexin: dosage nomograms for impaired renal function. Antimicrob. Agents Chemother. 1978, I't (2), 172-1 77.
SO SANH KET QUA DIEU TRj BENH PHINH DAI TRANG BAM SINH GliTA P H A U THUAT NOI SOI KET HOTP DOfiTNG QUA HAU MON VOI P H A U THUAT
QUA HAU MON DON THUAN TAI BENH VIEN NHI TRUNG UONG
TOM TAT
Nghien cu'u thu'c hien nham so sanh ket qua dieu tri benh phinh dai trang bam sinh (PDTBS) giiJa phau thuat ndi soi (PTNS) ket hop qua hau mon (QHM) va phau thult qua hau mdn ddn thuan tai Benh vien Nhi Trung u'dng. Ket qua cho thay thdi gian mo trung binh: PTNS 121 phiit, QHM 84 phut. Thdi gian nam vien sau mo trung binh: PTNS la 4 ± 0,1 ngay, QHM la 3 ± 0,2^ngay. Khdng co tir vong, bien chimg trong va sau md thap d ca hai nhdm. Ket qua chung ve chifc nSng dai tien danh gia theo phan loai Wingspread dat loai rat tdt va tdt cao d ca hai nhdm (78,8% va 77,2%). Viem rudt sau mo xay ra d: nhdm PTNS 5%, nhdm QHM 7%. Tieu tien binh thu'dng d tat ca benh nhan va tat ca cac benh nhan nam deu cd kha nang cu'dng du'dng vat vao buoi sang sau khi ngu day. Ket luan: Ket qua theo doi trung han cho thay ca hai phJOng phap PTNS va QHM deu an toan.
Bui Ou'c Hau*
bien chirng thap, tinh kha thi cao trong dieu trt benh POTBS. PTNS cd Uu diem hOn dUdng QHM la quan sat rd rang, giai quyet dUdc mpi the vd hach cao.
Tiy khoa: benh phinh dai trang bam sinh, phau thuat npi soi, du'dng qua hau mdn
SUMMARY
RESULT COMPARISION OF TREATMENT OF HIRSCHSPRUNG'S DISEASE BETWEEN PRIMARY
LAPAROSCOPtC-ASSISTED ENDORECTAL COLON PULL-THROUGH AND TRANSANAL IN
NATIONAL HOSPITAL OF PEDIATRICS The study was carried out to compare the outcomes of treatment of Hirschsprung's disease (HD) by the primary laparoscopic-assisted endorectal (LPS"
E) colon pull-through and transanal (TA) in Nationa' Hospital of Pediatrics. Results: The median operating Khoa Ngoai, Benh vien Nhi Trung ddng
Phan bien khoa hpc: GS.TS. Nguyin Thanh Liem 94
