title: Determination of methylmalonyl coenzyme A by ultra high-performance liquid chromatography tandem mass spectrometry for measuring propionyl coenzyme A carboxylase activity in patients with propionic acidemia.
abstract: Propionic acidemia (PA) is an inherited metabolic disease caused by low activity of propionyl coenzyme A (CoA) carboxylase (PCC), which metabolizes propionyl-CoA into methylmalonyl-CoA. Although many patients with PA have been identified by tandem mass spectrometry since the test was first included in neonatal mass screening in the 1990s, the disease severity varies. Thus, determining the specific level of PCC activity is considered to be helpful to grasp the severity of PA. We developed a new PCC assay method by the determination of methylmalonyl-CoA, which is formed by an enzyme reaction using peripheral lymphocytes, based on ultra high-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). With methylmalonyl-CoA concentrations of 0.05, 0.5, and 5μmol/L, the intra-assay coefficients of variation (CVs) were 8.2%, 8.7%, and 5.1%, respectively, and the inter-assay CVs were 13.6%, 10.5%, and 5.9%, respectively. The PCC activities of 20 healthy individuals and 6 PA patients were investigated with this assay. Methylmalonyl-CoA was not detected in one PA patient with a severe form of the disease, but the remaining PA patients with mild disease showed residual activities (3.3-7.8%). These results demonstrate that determination of PCC activity with this assay would be useful to distinguish between mild and severe cases of PA to help choose an appropriate treatment plan.
authors.affiliation: Department of Hospital Pharmacy, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan.
authors.forename: Kana
authors.initials: K
authors.lastname: Gotoh
authors.affiliation: Department of Pediatrics, Fujita Health University, Toyoake, Japan.
authors.forename: Yoko
authors.initials: Y
authors.lastname: Nakajima
authors.affiliation: Division of Neonatal Screening, National Center for Child health and Development, Tokyo, Japan.
authors.forename: Go
authors.initials: G
authors.lastname: Tajima
authors.affiliation: Department of Pediatrics and Child Health, Kurume University School of Medicine, Kurume, Japan; Research Institute of Medical Mass Spectrometry, Kurume University School of Medicine, Kurume, Japan.
authors.forename: Yoriko
authors.initials: Y
authors.lastname: Watanabe
authors.affiliation: Department of Hospital Pharmacy, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Hotta
authors.affiliation: Department of Clinical Pharmaceutics, Graduate of Medical Sciences, Nagoya City University, Nagoya, Japan.
authors.forename: Tomoya
authors.initials: T
authors.lastname: Kataoka
authors.affiliation: Department of Hospital Pharmacy, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Kawade
authors.affiliation: Aichi Medical College for Physical and Occupational Therapy, Kiyosu, Japan.
authors.forename: Naruji
authors.initials: N
authors.lastname: Sugiyama
authors.affiliation: Department of Pediatrics, Fujita Health University, Toyoake, Japan.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Ito
authors.affiliation: Department of Hospital Pharmacy, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan; Department of Clinical Pharmaceutics, Graduate of Medical Sciences, Nagoya City University, Nagoya, Japan.
authors.forename: Kazunori
authors.initials: K
authors.lastname: Kimura
authors.affiliation: Department of Hospital Pharmacy, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan. Electronic address: maeda@phar.nagoya-cu.ac.jp.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Maeda
mesh_terms: D000328:Adult; D016022:Case-Control Studies; D002648:Child; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005260:Female; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D057230:Limit of Detection; D016014:Linear Models; D008297:Male; D043586:Methylmalonyl-CoA Decarboxylase / Q000097:blood* / Q000378:metabolism*; D056693:Propionic Acidemia / Q000097:blood / Q000201:enzymology*; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*; D055815:Young Adult
chemical_list: D043586:Methylmalonyl-CoA Decarboxylase
keywords: Methylmalonyl coenzyme A; Propionic acidemia; Propionyl coenzyme A carboxylase; UPLC–MS/MS