Metabolism and bioactivation of the tricyclic antidepressant amitriptyline in human liver microsomes and human urine.
OBJECTIVE
Amitriptyline is a widely used tricyclic antidepressant, but the metabolic studies were conducted almost 20 years ago using high-performance liquid chromatography coupled with ultraviolet detector or radiolabeled methods.


RESULTS
First, multiple ion monitoring (MIM)- enhanced product ion (EPI) scan was used to obtain the diagnostic ions or neutral losses in human liver microsome incubations with amitriptyline. Subsequently, predicted multiple reaction monitoring (MRM)-EPI scan was used to identify the metabolites in human urine with the diagnostic ions or neutral losses. Finally, product ion filtering and neutral loss filtering were used as the data mining tools to screen metabolites. Consequently, a total of 28 metabolites were identified in human urine after an oral administration using LC-MS/MS.


CONCLUSIONS
An integrated workflow using LC-MS/MS was developed to comprehensively profile the metabolites of amitriptyline in human urine, in which five N-acetyl-l-cysteine conjugates were characterized as tentative biomarkers for idiosyncratic toxicity.
Tongji School of Pharmacy, Huazhong University of Science & Technology, 13 Hangkong Road, Wuhan 430030, Hubei, China.
Xin
X
Zhou
Tongji School of Pharmacy, Huazhong University of Science & Technology, 13 Hangkong Road, Wuhan 430030, Hubei, China.
Chang
C
Chen
Tongji School of Pharmacy, Huazhong University of Science & Technology, 13 Hangkong Road, Wuhan 430030, Hubei, China.
Fangrong
F
Zhang
Department of Pharmacy, Tongji Hospital of Tongji Medical College, Huazhong University of Science & Technology, 13 Hangkong Road, Wuhan, Hubei 430030, China.
Yang
Y
Zhang
Jiangxi University of Traditional Chinese Medicine, Nanchang 330006, China.
Yuling
Y
Feng
Jiangxi University of Traditional Chinese Medicine, Nanchang 330006, China.
Hui
H
Ouyang
Medical Research Center, Humanwell Healthcare (Group) Co., Ltd., 666 Gao xin Road, Wuhan, Hubei 430075, China.
Yong
Y
Xu
Tongji School of Pharmacy, Huazhong University of Science & Technology, 13 Hangkong Road, Wuhan 430030, Hubei, China.
Hongliang
H
Jiang
D000111:Acetylcysteine / Q000032:analysis / Q000378:metabolism / Q000652:urine; D000639:Amitriptyline / Q000032:analysis / Q000378:metabolism* / Q000652:urine*; D000929:Antidepressive Agents, Tricyclic / Q000032:analysis / Q000378:metabolism* / Q000652:urine*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006801:Humans; D053858:Metabolic Networks and Pathways; D008862:Microsomes, Liver / Q000378:metabolism*; D053719:Tandem Mass Spectrometry / Q000379:methods
D000929:Antidepressive Agents, Tricyclic; D000639:Amitriptyline; D000111:Acetylcysteine
GSH; LC–MS/MS; MIM-EPI; MRM-EPI; NAC; NLF; PIF; amitriptyline; biomarker; diagnostic ions