High-throughput solid-phase extraction for the determination of cimetidine in human plasma.
For the implementation and validation of an automated 'high-throughput' solid-phase extraction (SPE) system, using microtiter solid-phase technology and a pipetting robot, a SPE method previously validated manually for cimetidine in human plasma was adapted. Sample cleanup was performed by means of SPE using Microlute extraction plates in the 96-well format, each well filled with 50 mg of Varian C18 sorbent. Separation was performed by reversed-phase high-performance liquid chromatography (HPLC) with UV detection at 234 nm. The validated calibration range was from 0.100 to 5.00 mg/l, with an inaccuracy and imprecision below 20% at all concentration levels. Validation results on linearity, specificity, precision, accuracy and stability are shown and are found to be adequate. Cross-check analysis of samples from a clinical trial showed that there is a good correlation between results obtained by the automated method and results obtained by the manual method. The average sample preparation time for a technician decreased from approximately 4 min per sample to 0.6 min. A sample throughput of at least 160 samples per day can be achieved, the HPLC analysis time being the rate-limiting step.
Pharma Bio-Research International B.V., Zuidlaren, The Netherlands.
J
J
Hempenius
J
J
Wieling
J P
JP
Brakenhoff
F A
FA
Maris
J H
JH
Jonkman
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002927:Cimetidine / Q000097:blood*; D006801:Humans; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet
D002927:Cimetidine