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Simple Reversed-Phase HPLC Method with Spectrophotometric Detection for Measuring Acetaminophen-Protein Adducts in Rat Liver Samples
Joint Authors
Lau-Cam, Cesar A.
Acharya, Miteshkumar
Source
Issue
Vol. 2012, Issue 2012 (31 Dec. 2012), pp.1-6, 6 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2012-04-19
Country of Publication
Egypt
No. of Pages
6
Main Subjects
Natural & Life Sciences (Multidisciplinary)
Medicine
Information Technology and Computer Science
Abstract EN
A simple reversed-phase HPLC method for measuring hepatic levels of acetaminophen- (APAP-) protein adduct following an overdose of APAP was developed.
An aliquot of liver homogenate in phosphate-buffered saline pH 7.4 (PBS) was placed on a Nanosep centrifugal device, which was centrifuged to obtain a protein residue.
This residue was incubated with a solution of p-aminobenzoic acid (PABA), the internal standard, and bacterial protease in PBS, transferred to a Nanosep centrifugal device, and centrifuged.
A 100 μL portion of the filtrate was analyzed on a YMC-Pack ODS-AMQ C18 column, using 100 mM potassium dihydrogen phosphate-methanol-acetic acid (100 : 0.6 : 0.1) as the mobile phase, a flow rate of 1 mL/min, and photometric detection at 254 nm.
PABA and APAP-cystein-S-yl (APAP-Cys) eluted at ~14.7 min and 22.7 min, respectively.
Method linearity, based on on-column concentrations of APAP-Cys, was observed over the range 0.078–40 μg.
Recoveries of APAP-Cys from spiked blank liver homogenates ranged from ~83% to 91%.
Limits of detection and of quantification of APAP-Cys, based on column concentrations, were 0.06 μg and 0.14 μg, respectively.
RSD values for interday and intraday analyses of a blank liver homogenate spiked with APAP-Cyst at three levels were, in all cases, ≤1.0% and <1.5%, respectively.
The proposed method was found appropriate for comparing the antidotal properties of N-acetylcysteine and taurine in a rat model of APAP poisoning.
American Psychological Association (APA)
Acharya, Miteshkumar& Lau-Cam, Cesar A.. 2012. Simple Reversed-Phase HPLC Method with Spectrophotometric Detection for Measuring Acetaminophen-Protein Adducts in Rat Liver Samples. The Scientific World Journal،Vol. 2012, no. 2012, pp.1-6.
https://search.emarefa.net/detail/BIM-449328
Modern Language Association (MLA)
Acharya, Miteshkumar& Lau-Cam, Cesar A.. Simple Reversed-Phase HPLC Method with Spectrophotometric Detection for Measuring Acetaminophen-Protein Adducts in Rat Liver Samples. The Scientific World Journal No. 2012 (2012), pp.1-6.
https://search.emarefa.net/detail/BIM-449328
American Medical Association (AMA)
Acharya, Miteshkumar& Lau-Cam, Cesar A.. Simple Reversed-Phase HPLC Method with Spectrophotometric Detection for Measuring Acetaminophen-Protein Adducts in Rat Liver Samples. The Scientific World Journal. 2012. Vol. 2012, no. 2012, pp.1-6.
https://search.emarefa.net/detail/BIM-449328
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-449328