Modified method for determination of sulfur metabolites in plant tissues by stable isotope dilution-based liquid chromatography-electrospray ionization-tandem mass spectrometry

作者：Chang Ya Lan; Hsieh Chin Lin; Huang Yao Moan; Chiou Wen Liang; Kuo Yueh Hsiung; Tseng Mei Hwei^*

来源：Analytical Biochemistry, 2013, 442(1): 24-33.

DOI：10.1016/j.ab.2013.07.026

• Sulfur metabolites
• LC-MS/MS
• S-34 metabolic labeling

A wide variety of sulfur metabolites play important roles in plant functions. We have developed a precise and sensitive method for the simultaneous measurement of several sulfur metabolites based on liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) and S-34 metabolic labeling of sulfur-containing metabolites in Arabidopsis thaliana seedlings. However, some sulfur metabolites were unstable during the extraction procedure. Our proposed method does not allow for the detection of the important sulfur metabolite homocysteine because of its instability during sample extraction. Stable isotope-labeled sulfur metabolites of A. thaliana shoot were extracted and utilized as internal standards for quantification of sulfur metabolites with LC-MS/MS using S-adenosylmethionine (SAM), S-adenosylhomocysteine (SAH), methionine (Met), glutathione (GSH), and glutathione disulfide (GSSG) as example metabolites. These metabolites were detected using electrospray ionization in positive mode. Standard curves were linear (r(2) %26gt; 0.99) over a range of concentrations (SAM 0.01-2.0 mu M, SAH 0.002-0.10 mu M, Met 0.05-4.0 mu M, GSH 0.17-20.0 mu M, GSSG 0.07-20.0 mu M), with limits of detection for SAM, SAH, Met, GSH, and GSSG of 0.83, 0.67, 10, 0.56, and 1.1 nM, respectively: and the within-run and between-run coefficients of variation based on quality control samples were less than 8%.

• 出版日期2013-11-1
• 单位中国医科大学