Quantification of branched-chain amino acids in blood spots and plasma by liquid chromatography tandem mass spectrometry for the diagnosis of maple syrup urine disease

Individuals with maple syrup urine disease (MSUD) have an inherited metabolic disorder resulting in a deficiency in the branched-chain keto-acid dehydrogenase complex. As a result, these individuals have elevated concentrations of the branched-chain amino acids valine, isoluecine, allo-isoleucine, and leucine. MSUD presents in the first few days of life and progression may lead to irreversible intellectual disability, coma, cerebral edema, and death. However, early diagnosis and intervention can mitigate or eliminate many of the potential adverse effects. Consequently, it is important to develop techniques to screen for MSUD. We have developed an LC-MS/MS assay for the diagnosis of MSUD. The method is amenable to high-throughput formats due to the minimal sample prep required. The assay was shown to be robust, precise, and accurate. Finally, we identified and addressed some of the problems associated with working with bloodspots and implemented satisfactory approaches to overcoming these problems.