Assay for methylmalonyl coenzyme A mutase activity based on determination of succinyl coenzyme A by ultrahigh-performance liquid chromatography tandem mass spectrometry New Applications of Mass Spectrometry in Biomedicine

Kana Gotoh, Yoko Nakajima, Go Tajima, Yuji Hotta, Tomoya Kataoka, Yoshihiro Kawade, Naruji Sugiyama, Tetsuya Ito, Kazunori Kimura, Yasuhiro Maeda

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Abstract Methylmalonic acidemia (MMA) is an inherited metabolic disease. In this condition, metabolism from methylmalonyl coenzyme A (CoA) to succinyl-CoA is inhibited because of either low methylmalonyl-CoA mutase (MCM) activity or adenosylcobalamin deficiency owing to altered vitamin B12 metabolism. A high-precision assay for detecting MCM activity would facilitate not only MMA diagnosis but also the ability to determine the severity of MMA. We developed an MCM assay method based on ultrahigh-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) that involves the determination of succinyl-CoA, which is formed in an enzyme reaction, using peripheral lymphocytes. Using 0.05, 0.5, and 5 μmol/L succinyl-CoA, the intra-assay coefficient of variation (CV) was less than 5.2 % and the inter-assay CV was less than 8.7 %. The MCM activities of five healthy individuals and four patients were investigated with this assay. The MCM activities of the patients were very low in relation to those of healthy individuals. Together, these results show that the UPLC-MS/MS method is useful for a detailed MCM activity assay.

Original languageEnglish
Article number8753
Pages (from-to)5281-5286
Number of pages6
JournalAnalytical and Bioanalytical Chemistry
Volume407
Issue number18
DOIs
Publication statusPublished - 28-07-2015

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry

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