Column liquid chromatographic determination of clozapine and N-desmethylclozapine in human serum using solid-phase extraction

J Chromatogr B Biomed Appl. 1995 Nov 17;673(2):311-5. doi: 10.1016/0378-4347(95)00262-3.

Abstract

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.

MeSH terms

  • Antipsychotic Agents / blood*
  • Chromatography, High Pressure Liquid / methods*
  • Chromatography, High Pressure Liquid / statistics & numerical data
  • Clozapine / analogs & derivatives*
  • Clozapine / blood*
  • Humans
  • Regression Analysis
  • Sensitivity and Specificity

Substances

  • Antipsychotic Agents
  • norclozapine
  • Clozapine