Simultaneous determination of aceclofenac and diclofenac in human plasma by narrowbore HPLC using column-switching.
A fully automated narrowbore high performance liquid chromatography (HPLC) with column-switching was developed for the simultaneous determination of aceclofenac and diclofenac from human plasma samples. Plasma sample (100 microl) was directly introduced onto a Capcell Pak MF Ph-1 column (20 x 4 mm I.D.) where primary separation was occurred to remove proteins and concentrate target substances using acetonitrile potassium phosphate (pH 7, 0.1 M) (14:86, v/v). The drug molecules eluted from MF Ph-1 column were focused in an intermediate column (35 x 2 mm I.D.) by the valve switching step. The substances enriched in intermediate column were eluted and separated on the narrowbore phenyl hexyl column (100 x 2 mm I.D.) using acetonitrile-potassium phosphate (pH 7, 0.02M) (33:67, v/v) when the valve status was switched back to A position. The method showed excellent sensitivity (detection limit of 10 ng ml(-1)) with small volume of samples (100 microl), good precision and accuracy, and speed (total analysis time 17 min) without any loss in chromatographic efficiency. The response was linear (r2 > or = 0.999) over the concentration range of 50-10,000 ng ml(-1).
Lee
H S
HS
College of Pharmacy, Medical Resources Research Center, Wonkwang University, Iksan, South Korea. hslee@wonkwang.ac.kr
Jeong
C K
CK
Choi
S J
SJ
Kim
S B
SB
Lee
M H
MH
Ko
G I
GI
Sohn
D H
DH
D000328:Adult; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000097:blood* / Q000493:pharmacokinetics; D002851:Chromatography, High Pressure Liquid; D004008:Diclofenac / Q000031:analogs & derivatives* / Q000097:blood / Q000493:pharmacokinetics; D006801:Humans; D007202:Indicators and Reagents; D008297:Male; D015203:Reproducibility of Results
D000894:Anti-Inflammatory Agents, Non-Steroidal; D007202:Indicators and Reagents; D004008:Diclofenac; C056498:aceclofenac