An LC/ESI-MS/MS Method for Quantification of Hexanoylglycine in Three Different Urine Samples Within a Single Run Using Triplex 1-[(4- Diethylaminophenyl)carbonyl]piperazine Isotopologues

Liquid chromatography/electrospray ionization-tandem mass spectrometry (LC/ESI-MS/MS) has the great ability to accurately and precisely quantify various biomolecules, but there is a concern about its analysis time, especially during of high number samples. Sample-multiplexing in same injection promising strategy for reducing total time. This can be accomplished by derivatization multiple samples with isotopologous reagents. In this study, sample-triplex LC/ESI-MS/MS assay was developed quantifying urinary hexanoylglycine (HG), diagnostic marker medium-chain acyl-coenzyme A dehydrogenase deficiency, three different within single run. For purpose, 1-[(4-diethylaminophenyl)carbonyl]piperazine (DEAPPZ) isotopologues (2H0-, 2H3- 2H6-forms) were synthesized. When compared non-derivatization method, which analyzed one sample each run, time after pretreatment reduced 55% (390 min → 210 min) 30 also had an acceptable precision (intra- inter-assay precisions; ≤ 5.9% 9.1%, respectively) accuracy (91.7–97.1%). Thus, using DEAPPZ could successfully reduce HG quantification.