Development and Validation of a Novel Isotope Dilution-Ultraperformance Liquid Chromatography-Tandem Mass Spectrometry Method for Serum C-Peptide.

Autor:	Cho SE; Department of Endocrine Substance Analysis Center (ESAC), Yongin, Korea.; GC Labs, Yongin, Korea., Han J; Department of Endocrine Substance Analysis Center (ESAC), Yongin, Korea.; GC Labs, Yongin, Korea., You J; Department of Endocrine Substance Analysis Center (ESAC), Yongin, Korea.; GC Labs, Yongin, Korea., Lee JH; Department of Endocrine Substance Analysis Center (ESAC), Yongin, Korea.; GC Labs, Yongin, Korea., Yi A; Department of Endocrine Substance Analysis Center (ESAC), Yongin, Korea.; GC Labs, Yongin, Korea., Lee SG; GC Labs, Yongin, Korea., Lee EH; GC Labs, Yongin, Korea.
Jazyk:	angličtina
Zdroj:	Annals of laboratory medicine [Ann Lab Med] 2025 Jan 01; Vol. 45 (1), pp. 62-69. Date of Electronic Publication: 2024 Aug 23.
DOI:	10.3343/alm.2024.0072
Abstrakt:	Background: Mass spectrometry (MS) methods exhibit higher accuracy and comparability in measuring serum C-peptide concentrations than immunoassays. We developed and validated a novel isotope dilution-ultraperformance liquid chromatography-tandem MS (ID-UPLC-MS/MS) assay to measure serum C-peptide concentrations. Methods: Sample pretreatment involved solid-phase extraction, ion-exchange solid-phase extraction, and derivatization with 6-aminoquinolyl-N-hydroxysuccinimidylcarbamate (Cayman Chemical, Ann Arbor, Michigan, USA). We used an ExionLC UPLC system (Sciex, Framingham, MA, USA) and a Sciex Triple Quad 6500+ MS/MS system (Sciex) for electrospray ionization in positive-ion mode with multiple charge states of [M+3H]3+ and multiple reaction monitoring transitions. The total run time was 50 mins, and the flow rate was 0.20 mL/min. We evaluated the precision, trueness, linearity, lower limit of quantitation (LLOQ), carryover, and matrix effects. Method comparison with electrochemiluminescence immunoassay (ECLIA) was performed in 138 clinical specimens. Results: The intra- and inter-run precision coefficients of variation were <5% and the bias values for trueness were <4%, which were all acceptable. The verified linear interval was 0.050-15 ng/mL, and the LLOQ was 0.050 ng/mL. No significant carryover or matrix effects were observed. The correlation between this ID-UPLC-MS/MS method and ECLIA was good (R=0.995, slope=1.564); however, the ECLIA showed a positive bias (51.8%). Conclusions: The developed ID-UPLC-MS/MS assay shows acceptable performance in measuring serum C-peptide concentrations. This will be useful in situations requiring accurate measurement of serum C-peptide in clinical laboratories.
Databáze:	MEDLINE
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