Simultaneous determination of 6 beta-blockers, 3 calcium-channel antagonists, 4 angiotensin-II antagonists and 1 antiarrhythmic drug in post-mortem whole blood by automated solid phase extraction and liquid chromatography mass spectrometry. Method development and robustness testing by experimental design.
A method for the simultaneous determination of the beta-blockers atenolol, sotalol, metoprolol, bisoprolol, propranolol and carvedilol, the calcium-channel antagonists diltiazem, amlodipine and verapamil, the angiotensin-II antagonists losartan, irbesartan, valsartan and telmisartan, and the antiarrhythmic drug flecainide, in whole blood samples from forensic autopsies was developed. Sample clean-up was achieved by precipitation and solid phase extraction (SPE) with a mixed-mode column. Quantification was performed by reversed phase high performance liquid chromatography with positive electrospray ionization mass spectrometric detection (HPLC-MS). The method has been developed and robustness tested by systematically searching for satisfactory conditions using experimental designs including factorial and response surface designs. With the exception of amlodipine, the concentration limit of quantification (cLOQ) covered low therapeutic concentration levels for all the compounds. Within assay precisions and accuracies (bias) were 3.4-21% RSD and from -24 to 21% for the concentration range 1.00-5.00 microM, respectively. Between assay precisions were 4.4-28% RSD for the concentration range from 0.1 to 5 microM and recoveries varied from 9 to 103%. The method is used for determination of cardiovascular drugs in post-mortem whole blood samples from forensic autopsy cases.
Kristoffersen
Lena
L
Norwegian Institute of Public Health, Division of Forensic Toxicology and Drug Abuse, P.O. Box 4404 Nydalen, N-0403 Oslo, Norway. lena.kristoffersen@fhi.no
Øiestad
Elisabeth Leere
EL
Opdal
Mimi Stokke
MS
Krogh
Mette
M
Lundanes
Elsa
E
Christophersen
Asbjørg Solberg
AS
D000319:Adrenergic beta-Antagonists / Q000097:blood*; D057911:Angiotensin Receptor Antagonists; D000889:Anti-Arrhythmia Agents / Q000097:blood*; D001331:Automation; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D011180:Postmortem Changes; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D000319:Adrenergic beta-Antagonists; D057911:Angiotensin Receptor Antagonists; D000889:Anti-Arrhythmia Agents