Determination of penicillin-V in human plasma by high-performance liquid chromatography and solid-phase extraction.
A high-performance liquid chromatographic method has been developed for the determination of penicillin-V concentrations between 0.1 and 19 micrograms/ml in human plasma. Penicillin-V was isolated from plasma by solid-phase extraction on a C18/OH cartridge. The extracts were injected onto a reversed-phase HPLC system. A 125 x 4 mm C18 column was used to separate penicillin-V from its main metabolites, 5R- and 5S-penicilloic acid and endogenous compounds. The eluent consisted of 66% 0.02 M phosphoric acid buffer, to which tetrabutylammonium dihydrogenphosphate and 34% acetonitrile were added. The column effluent was monitored by ultraviolet spectrophotometry at 269 nm. Using this method, penicillin-V concentrations in plasma could be determined with an accuracy between -5.4 and 5.2% and a precision between 0.8 and 1.6%. The method has proved to be reliable and was used in bioavailability studies for the development of a new oral penicillin-V formulation.
Krauwinkel
W J
WJ
Biopharmaceutical Department, Yamanouchi Europe B.V., Leiderdorp, Netherlands.
Volkers-Kamermans
N J
NJ
D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D002951:Citrates / Q000737:chemistry; D004355:Drug Stability; D006801:Humans; D016014:Linear Models; D010397:Penicillanic Acid / Q000031:analogs & derivatives* / Q000032:analysis; D010404:Penicillin V / Q000097:blood* / Q000302:isolation & purification / Q000378:metabolism; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet
D002951:Citrates; C008782:penicilloic acid; D010397:Penicillanic Acid; D010404:Penicillin V