A novel fully validated LC-MS/MS method for quantification of pyridoxal-5'-phosphate concentrations in samples of human whole blood

Ghassabian, Sussan, Griffiths, Lyn and Smith, Maree T (2015) A novel fully validated LC-MS/MS method for quantification of pyridoxal-5'-phosphate concentrations in samples of human whole blood. Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, 1000 77-83. doi:10.1016/j.jchromb.2015.07.019

Attached Files (Some files may be inaccessible until you login with your UQ eSpace credentials)
Name Description MIMEType Size Downloads

Author Ghassabian, Sussan
Griffiths, Lyn
Smith, Maree T
Title A novel fully validated LC-MS/MS method for quantification of pyridoxal-5'-phosphate concentrations in samples of human whole blood
Journal name Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences   Check publisher's open access policy
ISSN 1873-376X
1570-0232
Publication date 2015-09-01
Year available 2015
Sub-type Article (original research)
DOI 10.1016/j.jchromb.2015.07.019
Open Access Status File (Author Post-print)
Volume 1000
Start page 77
End page 83
Total pages 7
Place of publication Amsterdam, Netherlands
Publisher Elsevier
Collection year 2016
Language eng
Abstract Quantification of pyridoxal-5′-phosphate (PLP) in biological samples is challenging due to the presence of endogenous PLP in matrices used for preparation of calibrators and quality control samples (QCs). Hence, we have developed an LC–MS/MS method for accurate and precise measurement of the concentrations of PLP in samples (20 μL) of human whole blood that addresses this issue by using a surrogate matrix and minimizing the matrix effect. We used a surrogate matrix comprising 2% bovine serum albumin (BSA) in phosphate buffer saline (PBS) for making calibrators, QCs and the concentrations were adjusted to include the endogenous PLP concentrations in the surrogate matrix according to the method of standard addition. PLP was separated from the other components of the sample matrix using protein precipitation with trichloroacetic acid 10% w/v. After centrifugation, supernatant were injected directly into the LC–MS/MS system. Calibration curves were linear and recovery was >92%. QCs were accurate, precise, stable for four freeze-thaw cycles, and following storage at room temperature for 17 h or at −80 °C for 3 months. There was no significant matrix effect using 9 different individual human blood samples. Our novel LC–MS/MS method has satisfied all of the criteria specified in the 2012 EMEA guideline on bioanalytical method validation.
Keyword Pyridoxal-5'-phosphate
LC-MS/MS
Method development
Method validation
Endogenous compounds
Bioanalysis
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2016 Collection
School of Pharmacy Publications
Centre for Integrated Preclinical Drug Development Publications
 
Versions
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 2 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 2 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Tue, 11 Aug 2015, 00:19:39 EST by System User on behalf of Scholarly Communication and Digitisation Service