Format

Send to

Choose Destination
Arch Biochem Biophys. 2014 Oct 1;559:3-11. doi: 10.1016/j.abb.2014.06.014. Epub 2014 Jun 23.

Improved high sensitivity analysis of polyphenols and their metabolites by nano-liquid chromatography-mass spectrometry.

Author information

1
The Targeted Metabolomics and Proteomics Laboratory, University of Alabama at Birmingham, Birmingham, AL, USA.
2
AB Sciex, Concord, Ontario, Canada.
3
Department of Pharmacology and Toxicology, University of Alabama at Birmingham, Birmingham, AL, USA; The Targeted Metabolomics and Proteomics Laboratory, University of Alabama at Birmingham, Birmingham, AL, USA; The O'Brien Acute Kidney Injury Center, University of Alabama at Birmingham, Birmingham, AL, USA.
4
Department of Pharmacology and Toxicology, University of Alabama at Birmingham, Birmingham, AL, USA; The Targeted Metabolomics and Proteomics Laboratory, University of Alabama at Birmingham, Birmingham, AL, USA; The O'Brien Acute Kidney Injury Center, University of Alabama at Birmingham, Birmingham, AL, USA. Electronic address: sbarnes@uab.edu.

Abstract

This study was conducted to assess the value of a high resolution, high mass accuracy time-of-flight analyzer in combination with nanoliquid chromatography for the analysis of polyphenols and their metabolites. The goal was to create a method that utilizes small volumes of biological fluids and provides a significant improvement in sensitivity compared with existing methods. Accordingly, nanoLC-MS and nanoLC-pseudo-multiple reaction monitoring (MRM) methods were developed that had a lower limit of quantification of 0.5 nM for several polyphenols and were linear over 2-3 orders of magnitude (R(2)>0.999). Using urine samples, the ability to observe and quantify polyphenols in such a complex biological fluid depended on much narrower mass windows (0.050 amu or less) on a TOF analyzer than those used on a quadrupole analyzer (0.7 amu). Although a greater selectivity was possible with the low mass resolution of a triple quadrupole instrument using the MRM approach, for the daidzein metabolite O-DMA, a chromatographically resolvable second peak could only be substantially reduced by using a 0.01 amu mass window. The advantage of a TOF analyzer for product ion data is that the whole MSMS spectrum is collected at high mass accuracy and MRM experiments are conducted in silico after the analysis.

KEYWORDS:

Mass accuracy; Nanoliquid chromatography–mass spectrometry; Polyphenols; Quantification; Sample volumes; Sensitivity

PMID:
24967696
PMCID:
PMC4143533
DOI:
10.1016/j.abb.2014.06.014
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Elsevier Science Icon for PubMed Central
Loading ...
Support Center