Simultaneous LC-MS/MS quantitation of acetaminophen and its glucuronide and sulfate metabolites in human dried blood spot samples collected by subjects in a pilot clinical study.
BACKGROUND
In support of a pilot clinical trial using acetaminophen as the model compound to assess dried blood spot (DBS) sampling as the method for clinical pharmacokinetic sample collection, a novel sensitive LC-MS/MS method was developed and validated for the simultaneous determination of acetaminophen and its major metabolites, acetaminophen glucuronide and sulfate, in human DBS samples collected by subjects via fingerprick.


RESULTS
The validated assay dynamic range was from 50.0 to 5000 ng/ml for each compound using a 1/8´´ (3-mm) disc punched from a DBS sample. Baseline separation of the three analytes was achieved to eliminate the possible impact of insource fragmentation of the conjugated metabolites on the analysis of the parent. The overall extraction efficiency was from 61.3 to 78.8% for the three analytes by direct extraction using methanol.


CONCLUSIONS
The validated method was successfully implemented in the pilot clinical study with the obtained pharmacokinetic parameters in agreement with the values reported in literature.
Department of Drug Metabolism & Pharmacokinetics, Novartis Institutes for Biomedical Research, One Health Plaza, East Hanover, NJ 07936, USA. wenkui.li@novartis.com
Wenkui
W
Li
John P
JP
Doherty
Kenneth
K
Kulmatycki
Harold T
HT
Smith
Francis Ls
FL
Tse
D000082:Acetaminophen / Q000031:analogs & derivatives* / Q000097:blood / Q000493:pharmacokinetics; D018712:Analgesics, Non-Narcotic / Q000097:blood* / Q000493:pharmacokinetics; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D059788:Dried Blood Spot Testing / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D057230:Limit of Detection; D000432:Methanol / Q000737:chemistry; D010865:Pilot Projects; D012015:Reference Standards; D052616:Solid Phase Extraction / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*
D018712:Analgesics, Non-Narcotic; D000082:Acetaminophen; C037386:acetaminophen glucuronide; C045667:acetaminophen sulfate ester; D000432:Methanol