Toxicological screening of human plasma by on-line SPE-HPLC-DAD: identification and quantification of basic drugs and metabolites.
An automated multi-analyte screening method for the identification and quantification of 92 drugs and metabolites based on on-line solid-phase extraction-high-performance liquid chromatography-diode array detection technique was developed and successfully validated. In addition, a database with 870 entries including UV-spectra, relative/retention times and response factors of toxicologically relevant compounds was created. Plasma samples (0.2 mL) were treated with methanol, diluted with buffer and on-line extracted (Strata X, 20 ×2 mm, 25 µm) at pH 9. Analytical separation was carried out on a Gemini NX column (150 ×4.6 mm, 3 µm) using gradient elution with acetonitrile-water (90:10,v/v) and 0.05 m potassium dihydrogen phosphate buffer (pH 2.3). Linear calibration curves with correlation coefficients ≥0.9950 were obtained for 78 analytes. As an additional benefit, the newly developed method allows the quantification of 42 analytes (e.g. antidepressants, neuroleptics and anticonvulsants) in a concentration range suitable for therapeutic drug monitoring. Limits of quantitation ranged from 0.02 mg/L (chlordiazepoxide) to 3.4 mg/L (mexiletine). Inter- and intra-day precisions of quality control samples (low/high) were better than 15% (zolpidem) and accuracy (bias) ranged from -11% (opipramol, venlafaxine) to 11% (venlafaxine, trazodone). Tests for carry-over and sample stability under different storage conditions were also performed and stability was adequate. Four cases of poisoning analysis are presented.
Berlin Institute of Technology, Department of Environmental Technology, Straße des 17. Juni 135, D-10623, Berlin, Germany.
Ludmila
L
Mut
Labor Berlin - Charité Vivantes GmbH, Department for Laboratory Medicine and Toxicology, Sylter Str. 2, D-13353, Berlin, Germany.
Thomas
T
Grobosch
Labor Berlin - Charité Vivantes GmbH, Department for Laboratory Medicine and Toxicology, Sylter Str. 2, D-13353, Berlin, Germany.
Torsten
T
Binscheck-Domaß
Berlin Institute of Technology, Department of Environmental Technology, Straße des 17. Juni 135, D-10623, Berlin, Germany.
Wolfgang
W
Frenzel
D000328:Adult; D001331:Automation; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D062313:Databases, Pharmaceutical; D005260:Female; D053593:Forensic Toxicology; D006801:Humans; D057230:Limit of Detection; D016014:Linear Models; D008297:Male; D008875:Middle Aged; D004364:Pharmaceutical Preparations / Q000097:blood* / Q000378:metabolism; D015203:Reproducibility of Results; D052616:Solid Phase Extraction / Q000379:methods*; D018675:Toxicity Tests / Q000379:methods*; D055815:Young Adult
D004364:Pharmaceutical Preparations
automated database search; automated solid-phase extraction; multi-analyte screening; systematic toxicological analysis; validation