Problems of Forensic Sciences 2003 Vol. 53 (LIII) 22-37
DETERMINATION OF DILTIAZEM IN POST-MORTEM BLOOD BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY
Marianna KISZKA, Roman MĄDRO
Chair and Department of Forensic Medicine, Medical Academy, Lublin
Streszczenie
The analytical conditions used in the determination of diltiazem (D) and deacetyldiltiazem (DAD) in post mortem blood using high-performance liquid chromatography (HPLC) are presented and a simple method of preparingDADis described. These xenobiotics were isolated from blood by liquid-liquid extraction (LLE) in a moderately alkaline medium with application of a mixture of dichloromethane and ether and determined with a liquid chromatograph (Gilson), using a Hypersil ODS (250 x 4 mm, 5 μm) column, a mobile phase of 1.5 ml/min flow (in an isocratic system of two pumps) consisting of acetonitrile and phosphate buffer at pH = 3 (0.025 M solution of phosphoric acid with addition of 6 ml of triethylamine per 1 litre) in proportions 25:75 and UV detection at 235 nm. Good separation of D and DAD from post mortem background was achieved and the average recovery from blood was 78% and 71% respectively. The level of detection of both xenobiotics was about 0.5 μg/ml (with a coefficient of variation CV within the range 2.6-9.1%). The method did allow us to determine concentrations that were twice as low (0.25 μg/ml), but at the cost of a decrease in precision (CV = 16.0-17.8%).
Słowa kluczowe
Diltiazem; Deacetyldiltiazem; Determination; High-performance liquid chromatography; Post-mortem blood.