A rapid LC-MS/MS assay for detection and monitoring of underivatized branched-chain amino acids in maple syrup urine disease

Hamed Piri-Moghadam, Alan Miller, Debra Pronger, Faye Vicente, Joel Charrow, Shannon Haymond, David C. Lin*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

2 Scopus citations


Introduction: Quantitation of the isomeric branched-chain amino acids (BCAA; valine, alloisoleucine, isoleucine, leucine) is a challenging task that typically requires derivatization steps or long runtimes if a traditional chromatographic method involving a ninhydrin ion pairing reagent is used. Objectives: To develop and perform clinical validation of a rapid, LC-MS/MS-based targeted metabolomics assay for detection and monitoring of underivatized BCAA in human plasma. Methods: Various columns and modes of chromatography were tested. The final optimized method utilized mixed mode chromatography with an Intrada column under isocratic condition. Sample preparation utilized the 96-well format. Briefly, extraction solvent containing the internal standard is added to 20 uL of sample, followed by shaking and positive pressure filtering, and the resulting extracted sample is analyzed. The assay was validated based on accepted quality standards (e.g., CLIA and CLSI) for clinical assays. Results: The method is linear over a wide range of concentrations, 2.0–1500 µM, with LOD of 0.60 µM and LOQ of 2.0 µM. The precision of the assay was 4–10% across analytes. The method was also validated against reference laboratories via blinded split-sample analysis and demonstrated good agreement with accuracy: 89–95% relative to the external group mean. Conclusion: We have developed a method that is accurate, rapid, and reliable for routine clinical testing of patient sample BCAA, which is used in the diagnosis and management of maple syrup urine disease (MSUD). The assay also has desirable characteristics, such as short run time, small sample volume requirement, simple sample preparation without the need for derivatization, and high throughput.

Original languageEnglish (US)
Pages (from-to)107-117
Number of pages11
JournalJournal of Mass Spectrometry and Advances in the Clinical Lab
StatePublished - Apr 2022


  • Branched-chain amino acid
  • Clinical assay
  • Maple syrup urine disease
  • Mass spectrometry
  • Quantitation

ASJC Scopus subject areas

  • Microbiology
  • Spectroscopy
  • Medical Laboratory Technology
  • Clinical Biochemistry


Dive into the research topics of 'A rapid LC-MS/MS assay for detection and monitoring of underivatized branched-chain amino acids in maple syrup urine disease'. Together they form a unique fingerprint.

Cite this