跳转至内容
Merck
  • Towards the chiral metabolomics: Liquid chromatography-mass spectrometry based DL-amino acid analysis after labeling with a new chiral reagent, (S)-2,5-dioxopyrrolidin-1-yl-1-(4,6-dimethoxy-1,3,5-triazin-2-yl)pyrrolidine-2-carboxylate, and the application to saliva of healthy volunteers.

Towards the chiral metabolomics: Liquid chromatography-mass spectrometry based DL-amino acid analysis after labeling with a new chiral reagent, (S)-2,5-dioxopyrrolidin-1-yl-1-(4,6-dimethoxy-1,3,5-triazin-2-yl)pyrrolidine-2-carboxylate, and the application to saliva of healthy volunteers.

Analytica chimica acta (2015-05-06)
Toshiki Mochizuki, Takahiro Takayama, Kenichiro Todoroki, Koichi Inoue, Jun Zhe Min, Toshimasa Toyo'oka
摘要

A novel triazine-type chiral derivatization reagent, i.e., (S)-2,5-dioxopyrrolidin-1-yl-1-(4,6-dimethoxy-1,3,5-triazin-2-yl) pyrrolidine-2-carboxylate (DMT-(S)-Pro-OSu), was developed for the highly sensitive and selective detection of chiral amines and amino acids by UPLC-MS/MS analysis. The enantiomers of amino acids were easily labeled with the reagents at room temperature within 40 min in an alkaline medium containing triethylamine. The diastereomers derived from proteolytic amino acids, except serine, were well separated under isocratic elution conditions by reversed-phase chromatography using an ODS column (Rs=1.2-9.0). DL-Serine was separated by use of an ADME column which has relatively higher polar surface than the conventional ODS column. The characteristic product ions, i.e., m/z 195.3 and m/z 209.3, were detected from all the diastereomers by the collision-induced dissociation of the protonated molecule. A highly sensitive detection on the amol-fmol level was obtained from the selected reaction monitoring (SRM) chromatogram. The chiral amines (e.g., adrenaline and noradrenaline) labeled with DMT-(S)-Pro-OSu were also well separated and sensitively detected by the present procedure. The method using DMT-(S)-Pro-OSu was used for the determination of DL-amino acids in the human saliva from healthy volunteers. Various L-amino acids were identified in the saliva. Furthermore, D-alanine (D-Ala) and D-proline (D-Pro) were also detected in relatively high concentrations (>5%). The ratio was higher in male saliva than in female saliva. However, the difference in the ratio of D-Ala for one day was not very high and the effect of foods and beverage seemed to be negligible. Based on the results using L-Ala-d3, the D-Ala in saliva seemed to be produced due to the racemization with some enzymes such as racemase. The racemization reaction was reversible, i.e., D-Ala-d3 was also racemized to L-Ala-d3 in saliva. Thus, care should be taken during the analysis of DL-amino acids in saliva. The present method using DMT-(S)-Pro-OSu may be applicable for the determination of chiral amine metabolomics, because the resulting derivatives produce the same product ions without relation to the compounds and show highly sensitive detection in the SRM mode of MS/MS. Consequently, DMT-(S)-Pro-OSu seems to be a useful chiral derivatization reagent for the determination of amines and amino acids in biological samples.

材料
货号
品牌
产品描述

Sigma-Aldrich
N-羟基丁二酰亚胺, 98%
Sigma-Aldrich
三乙基胺, BioUltra, ≥99.5% (GC)
Sigma-Aldrich
乙酸铵, for molecular biology, ≥98%
Sigma-Aldrich
乙酸铵 溶液, for molecular biology, 7.5 M
Sigma-Aldrich
甲醇, JIS special grade, ≥99.8%
Sigma-Aldrich
乙酸, for luminescence, BioUltra, ≥99.5% (GC)
Sigma-Aldrich
(−)-去甲肾上腺素, ≥98%, crystalline
Sigma-Aldrich
(R)-(+)-α-甲基苄胺, 98%
Sigma-Aldrich
乙胺 溶液, 66.0-72.0% in H2O
Sigma-Aldrich
乙酸铵, 99.999% trace metals basis
Sigma-Aldrich
乙酸, ≥99.5%, FCC, FG
Sigma-Aldrich
乙酸, natural, ≥99.5%, FG
Sigma-Aldrich
5α-雄甾烷-17β-醇-3-酮, ≥97.5%
Sigma-Aldrich
甲醇, SAJ first grade, ≥99.5%
Sigma-Aldrich
三乙基胺, ≥99.5%
Sigma-Aldrich
乙腈, anhydrous, 99.8%
Sigma-Aldrich
四氢呋喃, anhydrous, ≥99.9%, inhibitor-free
Sigma-Aldrich
甲醇, anhydrous, 99.8%
Sigma-Aldrich
四氢呋喃, anhydrous, contains 250 ppm BHT as inhibitor, ≥99.9%
Sigma-Aldrich
2-氯-4,6-二甲氧基-1,3,5-三嗪, 97%
Sigma-Aldrich
甲酸, ≥95%, FCC, FG
Sigma-Aldrich
乙胺, 99.0%
Sigma-Aldrich
乙酸, ≥99.7%
Sigma-Aldrich
乙胺, 97%
Sigma-Aldrich
乙酸, JIS special grade, ≥99.7%
Sigma-Aldrich
乙胺 溶液, 2.0 M in THF
Sigma-Aldrich
三乙基胺, ≥99%
Sigma-Aldrich
四氢呋喃, suitable for HPLC, contains no stabilizer
Sigma-Aldrich
1,2-二氯乙烷, anhydrous, 99.8%
Sigma-Aldrich
乙腈, electronic grade, 99.999% trace metals basis