Method comparison of HPLC-ninhydrin-photometry and UHPLC-PITC-tandem mass spectrometry for serum amino acid analyses in patients with complex congenital heart disease and controls

• DOI: 10.1007/s11306-020-01741-8
• Publication Date: 2020-12-15T04:46:32Z
• Authors: Miriam Michel; Christina Salvador; Verena Wiedemair; Mark Gordian Adam; Kai Thorsten Laser; Karl-Otto Dubowy; Andreas Entenmann; Daniela Karall; Ralf Geiger; Manuela Zlamy; Sabine Scholl-Bürgi
• Keywords: Ninhydrin; Coefficient of variation

Abstract Introduction Metabolomics studies are not routine when quantifying amino acids (AA) in congenital heart disease (CHD). Objectives Comparative analysis of 24 AA serum by traditional high-performance liquid chromatography (HPLC) based on ion exchange and ninhydrin derivatisation followed photometry ( PM ) with ultra-high-performance phenylisothiocyanate tandem mass spectrometry TMS ); interpretation findings CHD patients controls. Methods : Sample as above (total run time, ~ 119 min). AbsoluteIDQ® p180 kit assay (BIOCRATES Life Sciences AG, Innsbruck, Austria), which employs PITC derivatisation; separation analytes a Waters Acquity UHPLC BEH18 C18 reversed-phase column, using water acetonitrile 0.1% formic acid the mobile phases; quantification Triple-Stage Quadrupole spectrometer (Thermo Fisher Scientific, Waltham, MA) electrospray ionisation presence internal standards 8 Calculation coefficients variation (CV) (for precision), intra- interday accuracies, limits detection (LOD), (LOQ), mean concentrations. Results Both methods yielded acceptable results regard to precision (CV < 10% , 20% ), accuracies (< PM, 34% LOD, LOQ. For both Fontan controls concentrations differed significantly between methods, but patterns overall were parallel. Conclusion Serum differ analytical suitable for pattern recognition. TM S is time-saving alternative under physiological conditions well CHD. Trial registration number ClinicalTrials.gov Identifier NCT03886935, date March 27th, 2019 (retrospectively registered).