Column liquid chromatographic determination of clozapine and N-desmethylclozapine in human serum using solid-phase extraction.
A 0.5-ml aliquot of a serum sample, after the addition of a 50-microliters aliquot of a 5 microgram/ml solution of amoxapine as the internal standard, is vortex-mixed with 0.5 ml of acetonitrile and centrifuged. The supernatant is applied to a 1-ml BondElut C18 silica extraction column-conditioned with subsequent washings with 1 M HCL, methanol and water. After passing the sample at a slow rate, the column is washed twice with water and once with acetonitrile. The desired compounds are then eluted with a 0.25-ml aliquot of 35% perchloric acid-methanol (1:100, v/v). A 15 microliters aliquot of the eluate is injected onto a 150 x 4.6 mm I.D. column packed with 5-microns C8 silica particles and eluted at ambient temperature with a mobile phase of 0.1% tetramethylammonium perchlorate-acetonitrile (73:27, v/v) adjusted to pH 4.2 with 10% perchloric acid. The peaks are detected with an absorbance detector at 245 nm.
Gupta
R N
RN
Department of laboratory Medicine, St. Joseph's Hospital, Hamilton, Ont., Canada.
D014150:Antipsychotic Agents / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000706:statistics & numerical data; D003024:Clozapine / Q000031:analogs & derivatives* / Q000097:blood*; D006801:Humans; D012044:Regression Analysis; D012680:Sensitivity and Specificity
D014150:Antipsychotic Agents; C058272:norclozapine; D003024:Clozapine