Selective determination of amitriptyline and nortriptyline in human plasma by HPLC with ultraviolet and particle beam mass spectrometry.
A selective, sensitive, and reliable method was devised to determine concentrations of amitriptyline and its major metabolite, nortriptyline, in human plasma using high-performance liquid chromatography (HPLC) combined with UV and particle beam mass spectrometry (PBMS). Amitriptyline and nortriptyline were effectively extracted in a three-step solvent extraction procedure. Imipramine was used as the internal standard (IS). Amitriptyline, nortriptyline, and the IS were clearly separated by HPLC on a silica column with the mobile phase of acetonitrile-0.1 M ammonium acetate (94:6, v/v). The calibration curves were linear in the concentration range of 10-1000 ng/g for both compounds with UV and PBMS detections. The lower limits of detection were 5 ng/g for amitriptyline and 10 ng/g for nortriptyline with UV detection and 2 ng/g for amitriptyline and 5 ng/g for nortriptyline with PBMS detection. The absolute recoveries were 58% for amitriptyline and 47% for nortriptyline at a concentration of 50 ng/g. This method proved most useful in accurately identifying amitriptyline and nortriptyline in tissues from an autopsied individual.
Kudo
K
K
Department of Forensic Medicine, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
Jitsufuchi
N
N
Imamura
T
T
D000085:Acetates / Q000737:chemistry; D000097:Acetonitriles / Q000737:chemistry; D000639:Amitriptyline / Q000032:analysis / Q000097:blood* / Q000506:poisoning; D000929:Antidepressive Agents, Tricyclic / Q000032:analysis / Q000097:blood* / Q000506:poisoning; D001344:Autopsy; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D008099:Liver / Q000737:chemistry; D013058:Mass Spectrometry; D009661:Nortriptyline / Q000032:analysis / Q000097:blood*; D011041:Poisoning / Q000097:blood / Q000175:diagnosis; D012015:Reference Standards; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet
D000085:Acetates; D000097:Acetonitriles; D000929:Antidepressive Agents, Tricyclic; D000639:Amitriptyline; D009661:Nortriptyline; C018824:ammonium acetate; C032159:acetonitrile