alexa Improvements in the High-Performance Liquid Chromatography and Extraction Conditions for the Analysis of Oxidized Fatty Acids Using a Mixed-Mode Spin Column | Abstract
ISSN: 2329-6798

Modern Chemistry & Applications
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Research Article

Improvements in the High-Performance Liquid Chromatography and Extraction Conditions for the Analysis of Oxidized Fatty Acids Using a Mixed-Mode Spin Column

Takao Sanaki1, Takuji Fujihara1, Ryo Iwamoto2, Takeshi Yoshioka1*, Kenichi Higashino1, Toru Nakano1 and Yoshito Numata1
1Shionogi Innovation Center for Drug Discovery, Shionogi & Co., Ltd., Sapporo 001-0021, Japan
2Business-Academia-Collaborative Laboratory, Graduate School of Pharmaceutical Science, The University of Tokyo, Tokyo 113-0013, Japan
Corresponding Author : Takeshi Yoshioka
Shionogi Innovation Center for Drug Discovery
Shionogi & Co., Ltd., Sapporo 001-0021, Japan
Tel: +81-11-700-4713
Fax: +81-11-700-4716
E-mail:
[email protected]
Received July 30, 2015; Accepted August 07, 2015; Published August 12, 2015
Citation: Sanaki T, Fujihara T, Iwamoto R, Yoshioka T, Higashino K, et al. (2015) Improvements in the High-Performance Liquid Chromatography and Extraction Conditions for the Analysis of Oxidized Fatty Acids Using a Mixed-Mode Spin Column. Mod Chem appl 3:161. doi: 10.4172/2329-6798.1000161
Copyright: © 2015 Sanaki T, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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Abstract

Lipidomics by liquid chromatography/mass spectrometry has been used for a better understanding of the roles of oxidized fatty acids in the development of various diseases. However, further work is required to improve the sample preparation process and the peak tailing of cysteinyl-leukotrienes. In this study, we evaluated various mobile phases and extraction conditions. The addition of phosphoric acid to the mobile phase improved the peak tailing of cysteinyl-leukotrienes. The extraction conditions were also optimized by spin-column possessing an anion-exchange and reversed-phase properties. The extraction efficiency of the modified extraction system was examined using 62 lipids, and 13 deuterated lipids were investigated to evaluate matrix effects and recovery from mouse lung homogenate samples. Extraction efficiencies of ≥70% were obtained for almost all of the lipids. Good results with standard deviations of <15% were obtained for the matrix effects and recovery. Finally, the efficiency of our extraction method was compared with those of several conventional methods, and those of leukotriene C4 was improved significantly using our method. Moreover, the proportion of variance between our method and the conventional methods was >0.99 for all the lipids tested. This newly developed method therefore represents a powerful tool to analyze lipids.

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