TY - JOUR
T1 - 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
AU - Gotoh, Kana
AU - Nakajima, Yoko
AU - Tajima, Go
AU - Hotta, Yuji
AU - Kataoka, Tomoya
AU - Kawade, Yoshihiro
AU - Sugiyama, Naruji
AU - Ito, Tetsuya
AU - Kimura, Kazunori
AU - Maeda, Yasuhiro
N1 - Publisher Copyright:
© 2015 Springer-Verlag Berlin Heidelberg.
PY - 2015/7/28
Y1 - 2015/7/28
N2 - 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.
AB - 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.
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U2 - 10.1007/s00216-015-8753-8
DO - 10.1007/s00216-015-8753-8
M3 - Article
C2 - 26018627
AN - SCOPUS:84938093231
SN - 1618-2642
VL - 407
SP - 5281
EP - 5286
JO - Analytical and Bioanalytical Chemistry
JF - Analytical and Bioanalytical Chemistry
IS - 18
M1 - 8753
ER -