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Bias reduction in the quantitative analysis of a target analyte present in a limited quantity in human plasma using dual‐mode heart‐cutting two‐dimensional liquid chromatography coupled with isotope dilution mass spectrometry
Author(s) -
Hyung SeokWon,
Kim Byungjoo
Publication year - 2020
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.4831
Subject(s) - chemistry , analyte , chromatography , mass spectrometry , isotope dilution , matrix (chemical analysis) , analytical chemistry (journal) , liquid chromatography–mass spectrometry , triple quadrupole mass spectrometer , elution , high performance liquid chromatography , selected reaction monitoring , tandem mass spectrometry
Dual‐mode heart‐cutting two‐dimensional liquid chromatography (DMHC 2D‐LC) was applied to isotope dilution mass spectrometry (IDMS) to reduce the bias in the quantitative analysis of a target analyte present in a limited quantity in human plasma. Based on a Waters I‐Class LC system, the DMHC 2D‐LC system was operated in one‐ and two‐dimensional modes to facilitate the determination of heart‐cutting time and the efficient trapping of the target LC eluate. Experiments to determine the feasibility of coupling with IDMS were performed with triple quadrupole mass spectrometry using folic acid standards and/or 13 C 5 ‐folic acid. To validate the performance of the DMHC 2D‐LC/IDMS system on a complex sample, human plasma was analyzed for folic acid and the result was compared with that obtained using conventional single‐column LC. The total run time of the DMHC 2D‐LC system was 20 min, the same as that of the single‐column LC system. The peak profile of the spiked 13 C 5 ‐folic acid obtained with single‐column LC/MS was affected by matrix effects, but resolved with DMHC 2D‐LC/MS, thus improving the accuracy of the analysis. The DMHC 2D‐LC/IDMS system showed reliable performance in analyzing the target analyte in human plasma, eliminating matrix effects and saving analysis time.