title: A novel chiral stationary phase HPLC-MS/MS method to discriminate between enzymatic oxidation and auto-oxidation of phosphatidylcholine.
abstract: To elucidate the role of enzymatic lipid peroxidation in disease pathogenesis and in food deterioration, we recently achieved stereoselective analysis of phosphatidylcholine hydroperoxide (PCOOH) possessing 13S-hydroperoxy-9Z,11E-octadecadienoic acid (13(S)-9Z,11E-HPODE) using HPLC-MS/MS with a CHIRALPAK OP (+) column. Because enzymatic oxidation progresses concurrently with auto-oxidation, we need to distinguish them further. Here, we attempted such an analysis. First, we used lipoxygenase, linoleic acid, and lysophosphatidylcholine (LPC) to synthesize the enzymatic oxidation product 13(S)-9Z,11E-HPODE PC, and the auto-oxidation products 13(RS)-9Z,11E-HPODE PC and 13(RS)-9E,11E-HPODE PC, which were used as standards to test the ability of various columns to separate the enzymatic oxidation product from auto-oxidation products. Separation was achieved by connecting in series two columns with different properties: CHIRALPAK OP (+) and CHIRALPAK IB-3. The CHIRALPAK OP (+) column separated 13(R)-9Z,11E-HPODE PC and 13(S)-9Z,11E-HPODE PC, whereas CHIRALPAK IB-3 enabled separation of 13(S)-9Z,11E-HPODE PC and 13(RS)-9E,11E-HPODE PC. The results for the analysis of both enzymatically oxidized and auto-oxidized lecithin (an important phospholipid mixture in vivo and in food) indicate that our method would be useful for distinguishing enzymatic oxidation and auto-oxidation reactions. Such information will be invaluable for elucidating the involvement of PCOOH in disease pathogenesis and in food deterioration.
authors.affiliation: Food and Biodynamic Chemistry Laboratory, Graduate School of Agricultural Science, Tohoku University, Sendai, 981-8555, Japan.
authors.forename: Junya
authors.initials: J
authors.lastname: Ito
authors.affiliation: Food and Biodynamic Chemistry Laboratory, Graduate School of Agricultural Science, Tohoku University, Sendai, 981-8555, Japan. nkgw@m.tohoku.ac.jp.
authors.forename: Kiyotaka
authors.initials: K
authors.lastname: Nakagawa
authors.affiliation: Food and Biodynamic Chemistry Laboratory, Graduate School of Agricultural Science, Tohoku University, Sendai, 981-8555, Japan.
authors.forename: Shunji
authors.initials: S
authors.lastname: Kato
authors.affiliation: Laboratory of Applied Bioorganic Chemistry, Graduate School of Agricultural Science, Tohoku University, Sendai, 981-8555, Japan.
authors.forename: Takafumi
authors.initials: T
authors.lastname: Hirokawa
authors.affiliation: Laboratory of Applied Bioorganic Chemistry, Graduate School of Agricultural Science, Tohoku University, Sendai, 981-8555, Japan.
authors.forename: Shigefumi
authors.initials: S
authors.lastname: Kuwahara
authors.affiliation: Tsukishima Foods Industry Co., Ltd., Tokyo, 134-8520, Japan.
authors.forename: Toshiharu
authors.initials: T
authors.lastname: Nagai
authors.affiliation: Food and Biodynamic Chemistry Laboratory, Graduate School of Agricultural Science, Tohoku University, Sendai, 981-8555, Japan.|New Industry Creation Hatchery Center (NICHe), Tohoku University, Sendai, 980-8579, Japan.
authors.forename: Teruo
authors.initials: T
authors.lastname: Miyazawa
mesh_terms: D005591:Chemical Fractionation / Q000295:instrumentation / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D054709:Lecithins / Q000737:chemistry; D019787:Linoleic Acid / Q000737:chemistry; D008041:Linoleic Acids / Q000737:chemistry / Q000302:isolation & purification*; D015227:Lipid Peroxidation; D008054:Lipid Peroxides / Q000737:chemistry / Q000302:isolation & purification*; D008084:Lipoxygenase / Q000737:chemistry; D008244:Lysophosphatidylcholines / Q000737:chemistry; D010713:Phosphatidylcholines / Q000737:chemistry* / Q000302:isolation & purification; D013025:Glycine max / Q000737:chemistry / Q000378:metabolism; D013237:Stereoisomerism; D053719:Tandem Mass Spectrometry / Q000295:instrumentation / Q000379:methods*
chemical_list: D054709:Lecithins; D008041:Linoleic Acids; D008054:Lipid Peroxides; D008244:Lysophosphatidylcholines; D010713:Phosphatidylcholines; C057590:phosphatidylcholine hydroperoxide; C038649:13-hydroperoxy-9,11-octadecadienoic acid; D019787:Linoleic Acid; D008084:Lipoxygenase
keywords: Chiral column; Chiral stationary phase; LC-MS/MS; Lipid oxidation; Phosphatidylcholine hydroperoxide