Edogawa-ku, Japan
Edogawa-ku, Japan

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Ito J.,Tohoku University | Nakagawa K.,Tohoku University | Kato S.,Tohoku University | Hirokawa T.,Tohoku University | And 3 more authors.
Journal of Chromatography A | Year: 2015

Increasing evidence suggests that phospholipid peroxidation plays important roles in the pathogenesis of various diseases, such as atherosclerosis. With regard to the biochemical processes that initiate phospholipid peroxidation in vivo, enzymatic conversion of phosphatidylcholine to phosphatidylcholine hydroperoxide (PCOOH) by lipoxygenase (LOX) may play a crucial role. This will become clear if we can analyze PCOOH bearing hydroperoxy fatty acids with S-stereoconfiguration. In this study, we therefore attempted such an analysis. Initially, we used LOX, linoleic acid and Lyso phosphatidylcholine, and synthesized PCOOH bearing 13. S-hydroperoxy-9. Z,11. E-octadecadienoic acid (13(. S)-9. Z,11. E-HPODE). PCOOH bearing racemic 13-9. Z,11. E-HPODE was also prepared. We used liquid chromatography equipped with CHIRALPAK OP (+) (poly (. o-pyridyl diphenylmethacrylate) coated on silica), a UV detector and a quadrupole-time-of-flight mass spectrometer, and achieved diastereomer separation of PCOOH stereoisomers with excellent resolution and peak shape. This is the first study reporting the diastereomer separation of PCOOH. The present method will be beneficial in developing a better understanding of the biochemical processes that initiate oxidative stress (PCOOH formation) in vivo, which may lead to further elucidation of the involvement of PCOOH in the development of diseases. In addition to clinical applications, the present method may also be effective in the evaluation of enzymatic oxidative food deterioration. © 2015 Elsevier B.V.


Increasing evidence suggests that phospholipid peroxidation plays important roles in the pathogenesis of various diseases, such as atherosclerosis. With regard to the biochemical processes that initiate phospholipid peroxidation in vivo, enzymatic conversion of phosphatidylcholine to phosphatidylcholine hydroperoxide (PCOOH) by lipoxygenase (LOX) may play a crucial role. This will become clear if we can analyze PCOOH bearing hydroperoxy fatty acids with S-stereoconfiguration. In this study, we therefore attempted such an analysis. Initially, we used LOX, linoleic acid and Lyso phosphatidylcholine, and synthesized PCOOH bearing 13S-hydroperoxy-9Z,11E-octadecadienoic acid (13(S)-9Z,11E-HPODE). PCOOH bearing racemic 13-9Z,11E-HPODE was also prepared. We used liquid chromatography equipped with CHIRALPAK OP (+) (poly (o-pyridyl diphenylmethacrylate) coated on silica), a UV detector and a quadrupole-time-of-flight mass spectrometer, and achieved diastereomer separation of PCOOH stereoisomers with excellent resolution and peak shape. This is the first study reporting the diastereomer separation of PCOOH. The present method will be beneficial in developing a better understanding of the biochemical processes that initiate oxidative stress (PCOOH formation) in vivo, which may lead to further elucidation of the involvement of PCOOH in the development of diseases. In addition to clinical applications, the present method may also be effective in the evaluation of enzymatic oxidative food deterioration.


Gotoh N.,Tokyo University of Marine Science and Technology | Wada S.,Tokyo University of Marine Science and Technology | Nagai T.,Tsukishima Foods Industry Co.
Lipid Technology | Year: 2011

A recycle HPLC system equipped with a polysaccharide-based chiral column was used for the enantiomeric separation of asymmetric triacylglycerols (TAGs). When the chiral columns were screened by the resolution of 1,2-dipalmitoyl-3-oleoyl-rac-glycerol separation was achieved only with the cellulose tris- (3,5-dimethylphenylcarbamate) chiral selector. The resolution of other TAG enantiomers was also examined, and 1,2-dioleoyl-3-palmitoyl-rac-glycerol, 1,2-dipalmitoyl-3-linoleoyl-rac-glycerol, 1,2-dipalmitoyl-3-eicosapentaenoyl-rac-glycerol, 1,2-dipalmitoyl-3-docosahexaenoyl-rac-glycerol, and 1,2-docosahexaenoyl-3-palmitoyl-rac-glycerol were resolved into their respective enantiomers. However, neither 1,2-dioleoyl-3-linoleoyl-rac-glycerol, consisting of only unsaturated fatty acids, nor 1,2-dipalmitoyl-3-stearoyl-rac-glycerol, consisting of only saturated fatty acids, was resolved. In addition, 1,2-eicosapentaenoyl-3-palmitoyl-rac-glycerol was not resolved clearly, even in the recycle runs. These results suggest that asymmetric TAGs having both a palmitic acid moiety and an unsaturated fatty acid moiety at the sn-1 or sn-3 positions might be resolved on a cellulose tris- (3,5-dimethylphenylcarbamate) chiral column. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.


Gotoh N.,Tokyo University of Marine Science and Technology | Matsumoto Y.,Tokyo University of Marine Science and Technology | Nagai T.,Tsukishima Foods Industry Co. | Mizobe H.,Tsukishima Foods Industry Co. | And 6 more authors.
Food Chemistry | Year: 2011

The distribution of fatty acid species at the (sn-1, 3) position or the (sn-2) position of triacylglycerol (TAG) in natural fats and oils has already been analysed by many researchers and several interesting results have been reported. However, most of these reports only focused on the distribution of fatty acids at the or positions in TAG, and did not take account of the combination of fatty acids in the TAG, i.e., the TAG positional isomers. In this study, the actual ratios of TAG positional isomer pairs, consisting of palmitic acid and highly unsaturated fatty acid (HUFA) such as DHA or EPA, in fish and marine mammals were investigated using a high-performance liquid chromatography/atmospheric pressure chemical ionisation-mass spectrometry (HPLC/APCI-MS) system equipped with tandem jointed non-endcapped polymeric ODS columns. The results show that for combinations of DHA or EPA with two palmitic acids in the TAG of marine mammals, binding was almost all at the α position. In contrast, binding of DHA or EPA was mainly at the β position in fish. The preferred DHA and EPA positions in TAG were the same in the same marine mammal or fish. The binding position tendency of HUFA in TAG positional isomers consisting of two HUFAs and one palmitic acid was the same as that for combinations of one HUFA and two palmitic acids. These results were interpreted as showing that the preferred fatty acid species of sn-glycerol-3-phosphate acyltransferase and 1-acyl-sn-glycerol-3-phosphate acyltransferase in marine mammals are different to those in fish and other animals, or that diacylglycerol acyltransferase in marine mammals favours 1,2-dipalmitoyl-sn-glycerol formed from 1,2-dipalmitoyl-sn-glycerol-3-phosphatidate if HUFA is the reaction substrate. © 2011 Elsevier Ltd. All rights reserved.


Lee J.W.,Osaka University | Nagai T.,Tsukishima Foods Industry Co. | Gotoh N.,Tokyo University of Marine Science and Technology | Fukusaki E.,Osaka University | Bamba T.,Osaka University
Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences | Year: 2014

In this study, supercritical fluid chromatography (SFC) coupled with triple quadrupole mass spectrometry was applied to the profiling of several regioisomeric triacylglycerols (TAGs). SFC conditions (column, flow rate, modifier) were optimized for the effective separation of TAGs. In the column test, a triacontyl (C30) silica gel reversed-phase column was selected to separate TAG regioisomers. Multiple reaction monitoring was used to selectively quantify each TAG. Then, the method was used to perform detailed characterization of a diverse array of TAGs in palm and canola oils. Seventy TAGs (C46:0-C60:2) of these oils were successfully analyzed as a result, and twenty isomeric TAG pairs were separated well. In particular, this method provided the fast and high resolution separation of six regioisomeric TAG pairs (PPLn/PLnP, PPL/PLP, PPO/POP, SPLn/SLnP, SPO/SOP, SSO/SOS-stearic acid (S, 18:0), oleic acid (O, 18:1), linoleic acid (L, 18:2), linolenic acid (Ln, 18:3), palmitic acid (P, 16:0)) in a short time (50. min) as compared to high performance liquid chromatography. We were able to demonstrate the utility of this method for the analysis of regioisomeric TAGs in edible oils. © 2014 Elsevier B.V.


Yoshinaga K.,Tsukishima Foods Industry Co. | Asanuma M.,Tokyo University of Marine Science and Technology | Mizobe H.,Tsukishima Foods Industry Co. | Kojima K.,Tsukishima Foods Industry Co. | And 3 more authors.
Food Chemistry | Year: 2014

In this study, the characterisation of all cis- and trans-octadecenoic acid (C18:1) positional isomers in partially hydrogenated vegetable oil (PHVO) and milk fat, which contain several cis- and trans-C18:1 positional isomers, was achieved by gas chromatography-flame ionisation detector equipped with a highly polar ionic liquid capillary column (SLB-IL111). Prior to analysis, the cis- and trans-C18:1 fractions in PHVO and milk fat were separated using a silver-ion cartridge. The resolution of all cis-C18:1 positional isomers was successfully accomplished at the optimal isothermal column temperature of 120 °C. Similarly, the positional isomers of trans-C18:1, except for trans-6-C18:1 and trans-7-C18:1, were separated at 120 °C. The resolution of trans-6-C18:1 and trans-7-C18:1 isomers was made possible by increasing the column temperature to 160 °C. This analytical method is suitable for determining the cis- and trans-C18:1 positional isomers in edible fats and oils. © 2014 Elsevier Ltd. All rights reserved.


Nagai T.,Tsukishima Foods Industry Co. | Mizobe H.,Tsukishima Foods Industry Co. | Otake I.,Tsukishima Foods Industry Co. | Ichioka K.,Tsukishima Foods Industry Co. | And 5 more authors.
Journal of Chromatography A | Year: 2011

In our previous studies, we employed recycle HPLC for the separation of triacylglycerol (TAG)-positional isomers (PIs). In this study, a recycle HPLC system equipped with a polysaccharide-based chiral column was applied to the enantiomeric separation of some asymmetric TAGs having straight-chain C16-C18 acyl residues. As a result, 1,2-dipalmitoyl-3-oleoyl-rac-glycerol (. rac-PPO), 1,2-dioleoyl-3-palmitoyl-rac-glycerol (. rac-OOP), and 1,2-dipalmitoyl-3-linoleoyl-rac-glycerol (. rac-PPL) were resolved into their respective enantiomers. However, neither 1,2-dioleoyl-3-linoleoyl-rac-glycerol (. rac-OOL), consisting of only unsaturated fatty acids, nor 1,2-dipalmitoyl-3-stearoyl-rac-glycerol (. rac-PPS), consisting of only saturated fatty acids, was resolved. These results suggest that the asymmetric TAGs, used in this study, having both a palmitic acid moiety and an oleic acid (or a linoleic acid) moiety at the sn-1 or sn-3 positions are resolved by the chiral column. This new chiral separation method can be used in combination with atmospheric pressure chemical ionization mass spectrometry to determine the sn-OOP/. sn-POO ratio in palm oil. This method is applicable for the chiral separation of asymmetric TAGs in palm oil. © 2011 Elsevier B.V.


Yoshinaga K.,Tsukishima Foods Industry Co. | Yoshinaga K.,Tokyo University of Marine Science and Technology | Sasaki K.,Tokyo University of Marine Science and Technology | Watanabe H.,Kochi University | And 9 more authors.
Journal of Nutritional Biochemistry | Year: 2015

The present study investigated the effects of binding position of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) to triacylglycerol (TAG) on lipid metabolism in C57BL/6J mice. Mice were treated with pure TAG positional isomers, including 1,2(2,3)-dipalmitoyl-3(1)-eicosapentaenoyl glycerol, 1,3-dipalmitoyl-2-eicosapentaenoyl glycerol, 1,2(2,3)-dipalmitoyl-3(1)-docosahexaenoyl glycerol, and 1,3-dipalmitoyl-2-docosahexaenoyl glycerol. Compared to DHA bound to the α-position of TAG, DHA bound to the β-position more effectively inhibited fatty acid synthetic enzymes and cholesterol-metabolism enzymes and thus reduced TAG and cholesterol concentrations in the serum and liver. EPA bound to the α-position of TAG, but not EPA bound to the β-position of TAG, significantly decreased hepatic cholesterol concentrations. Additionally, EPA bound to the α-position of TAG increased the ratio of PGI2 to TXA2 to a higher degree than EPA bound to the β-position. These results suggested that the binding position of EPA and DHA to TAG affected TAG and cholesterol metabolism as well as eicosanoid production in C57BL/6J mice. © 2015 Elsevier Inc.


PubMed | Tsukishima Foods Industry Co.
Type: Journal Article | Journal: Journal of oleo science | Year: 2015

Bovine milk fat (BMF) is composed of triacylglycerols (TAG) rich in palmitic acid (P), oleic acid (O), and short-chain or medium-chain fatty acids (SCFAs or MCFAs). The composition and binding positions of the fatty acids on the glycerol backbone determine their physical and nutritional properties. SCFAs and MCFAs are known to characteristically bind to the sn-3 position of the TAGs in BMF; however, there are very few non-destructive analyses of TAG enantiomers binding the fatty acids at this position. We previously reported a method to resolve the enantiomers of TAGs, binding both long-chain saturated fatty acid and unsaturated fatty acid at the sn-1 and 3 positions, in palm oil, fish oil, and marine mammal oil using chiral HPLC. Here, we further developed a method to resolve several TAG enantiomers containing a dipalmitoyl (PP) glycerol backbone and one SCFA (or MCFA) in BMF. We revealed that the predominant TAG structure in BMF was homochiral, such as 1,2-dipalmitoyl-3-butyroyl-sn-glycerol. This is the first quantitative determination of many TAG enantiomers, which bind to a SCFA or MCFA, in BMF was evaluated simultaneously. Furthermore, the results indicated that the amount ratios of the positional isomers and enantiomers of TAGs consisting of a dipalmitoyl (PP) glycerol backbone and SCFA (or MCFA), resembled the whole TAG structures containing the other diacylglycerol backbones consisting of P, O, myristic acid, and/or stearic acid in BMF.


The positional distributions of fatty acids (FAs) in milk fat containing short- and medium-chain FAs were analyzed by sn-1(3)-selective transesterification of triacylglycerols (TAGs) with ethanol using immobilized Candida antarctica lipase B (CALB), in a collaborative study conducted by 10 laboratories. The mean C4:0, C6:0, and C8:0 FA contents, when analyzed as propyl esters (PEs) using gas chromatography (GC) with a DB-23 capillary column, were found to be 3.0, 2.0, and, 1.3 area%, respectively. Their reproducibility standard deviations were 0.33, 0.18, and 0.19, respectively. The mean C4:0, C6:0, and C8:0 contents at the sn-2 position were 0.3, 0.4, and 1.0 area%, respectively. Their reproducibility standard deviations were 0.17, 0.11, and 0.19, respectively. The reproducibility standard deviations of C4:0, C6:0, and C8:0 FAs at the sn-2 position were either the same as or smaller than those for milk fat, although the FA contents at the sn-2 position were smaller than those in the milk fat. Therefore, it was concluded that the CALB method for estimating the regiospecific distribution is applicable to TAGs containing short- and medium-chain FAs. When estimating the short-chain (SC) FA contents in fats and oils by GC, it is better to analyze SCFAs as PEs or butyl esters, and not as methyl esters, in order to prevent loss of SCFAs during the experimental procedure because of their volatility and water solubility. This study also revealed that the stationary phase of the GC capillary column affected the flame ionization detector (FID) response of SCFAs. The theoretical FID correction factor (MWFA / active carbon number / atomic weight of carbon) fitted well with the actual FID responses of C4:0-C12:0 FAs when they were analyzed as PEs using a DB-23 column; however, this was not the case when the GC analysis was performed using wax-type columns.

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