用于测定魁蚶中D-天冬氨酸甲基转移酶活性的液相色谱-电喷雾电离-串联质谱分析法

Liquid chromatography-electrospray ionization-tandem mass spectrometric assay for d-aspartate -methyltransferase activity in ark shells.

作者信息

Shibata Kimihiko, Hagiya Saho, Okawara Rena, Abe Katsumasa, Takahashi Shouji, Kera Yoshio

机构信息

Department of Applied Chemistry and Biochemistry, National Institute of Technology (KOSEN), Fukushima College, Iwaki, Fukushima, Japan.

Department of Bioengineering, Nagaoka University of Technology, Nagaoka, Niigata, Japan.

出版信息

Biosci Biotechnol Biochem. 2020 Mar;84(3):500-506. doi: 10.1080/09168451.2019.1689094. Epub 2019 Nov 6.

DOI:10.1080/09168451.2019.1689094

PMID:31694479

Abstract

A liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) method for the separation and quantification of the enantiomers of -methylaspartate and -methylglutamate, after derivatization with -(5-fluoro-2,4-dinitrophenyl)-L-leucinamide was established. The time required for the LC-ESI-MS/MS analysis was within 20 min and the detection limit was approximately 10 fmol per injection, demonstrating that this method can be used for the rapid determination of D-aspartate -methyltransferase activity in the ark shell clam . NMDA: -methyl-D-aspartate; NMLA: -methyl-L-aspartate; NMDG: -methyl-D-glutamate; NMLG: -methyl-L-glutamate; NMA: -methylaspartate; NMG: -methylglutamate; HPLC: high-performance liquid chromatography; SAM: -adenosyl-L-methionine; OPA: -phthalaldehyde; LC-ESI-MS/MS: liquid chromatography-electrospray ionization-tandem mass spectrometry; FDLA: -(5-fluoro-2,4-dinitrophenyl)-L-leucinamide; FDAA: -(5-fluoro-2,4-dinitrophenyl)-L-alaninamide; ESI: electrospray ionization; LC-ESI-MS: liquid chromatography-electrospray ionization-mass spectrometry; MS/MS: tandem mass spectrometry.

摘要

建立了一种液相色谱 - 电喷雾电离 - 串联质谱（LC - ESI - MS/MS）方法，用于在使用α-(5 - 氟 - 2,4 - 二硝基苯基)-L - 亮氨酰胺衍生化后分离和定量α-甲基天冬氨酸和α-甲基谷氨酸的对映体。LC - ESI - MS/MS分析所需时间在20分钟以内，检测限约为每次进样10飞摩尔，表明该方法可用于快速测定贻贝中D - 天冬氨酸α-甲基转移酶的活性。NMDA：α-甲基 - D - 天冬氨酸；NMLA：α-甲基 - L - 天冬氨酸；NMDG：α-甲基 - D - 谷氨酸；NMLG：α-甲基 - L - 谷氨酸；NMA：α-甲基天冬氨酸；NMG：α-甲基谷氨酸；HPLC：高效液相色谱；SAM：S - 腺苷 - L - 甲硫氨酸；OPA：邻苯二甲醛；LC - ESI - MS/MS：液相色谱 - 电喷雾电离 - 串联质谱；FDLA：α-(5 - 氟 - 2,4 - 二硝基苯基)-L - 亮氨酰胺；FDAA：α-(5 - 氟 - 2,4 - 二硝基苯基)-L - 丙氨酰胺；ESI：电喷雾电离；LC - ESI - MS：液相色谱 - 电喷雾电离 - 质谱；MS/MS：串联质谱。