Premium
Ultra‐pressure liquid chromatography/tandem mass spectrometry targeted profiling of arachidonic acid and eicosanoids in human colorectal cancer
Author(s) -
Mal Mainak,
Koh Poh Koon,
Cheah Peh Yean,
Chan Eric Chun Yong
Publication year - 2011
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.4926
Subject(s) - chemistry , formic acid , chromatography , selected reaction monitoring , tandem mass spectrometry , arachidonic acid , high performance liquid chromatography , liquid chromatography–mass spectrometry , mass spectrometry , biochemistry , enzyme
Cumulative evidence shows that eicosanoids such as prostaglandins, leukotrienes, thromboxanes and hydroxy eicosatetraenoic acids play an important role in associating inflammation with human colorectal cancer (CRC). In this study an ultra‐pressure liquid chromatography/tandem mass spectrometry (UPLC/MS/MS) method was developed and validated for the targeted profiling of eight relevant eicosanoids and the major metabolic precursor, arachidonic acid (AA), in human colon. Multiple reaction monitoring (MRM) experiments were performed in negative electrospray ionization mode. The metabolites were separated using a C 18 column consisting of 1.7 µm ethylene‐bridged hybrid particles (100 × 2.1 mm i.d.) and gradient elution (50 to 95% of solvent B) with a mobile phase comprising water (0.1% formic acid) [solvent A] and acetonitrile (0.1% formic acid) [solvent B] at a flow rate of 0.4 mL/min. The analysis time for each sample was 5.5 min. Our UPLC/MS/MS method demonstrated satisfactory validation results in terms of selectivity, sensitivity, matrix effect, linearity, extraction efficiency, intra‐ and inter‐day precision, accuracy and autosampler stability. The method was applied for the clinical profiling of matched pairs of cancerous and normal colon mucosae obtained from eight colorectal cancer patients. Endogenous levels of AA and selected eicosanoids such as prostaglandin E 2 (PGE 2 ), prostacyclin (PGI 2 ) [assayed as its stable hydrolytic product 6‐keto‐prostaglandin 1α (6‐k PGF 1α )] and 12‐hydroxy‐5 Z ,8 Z ,10 E ,14 Z ‐eicosatetraenoic acid (12‐HETE) were found to be significantly different ( p <0.05; paired t‐test) between cancerous and normal mucosae. Copyright © 2011 John Wiley & Sons, Ltd.