Qualititative analysis of 5-methyltertrahydrofolate in human serum by ID-LC-MS/MS
10.13602/j.cnki.jcls.2018.03.01
- VernacularTitle:同位素稀释液相色谱串联质谱法测定人血清中5-甲基四氢叶酸
- Author:
Xu PENG
1
;
Yingshu ZOU
;
Zongbing YANG
;
Jun WANG
Author Information
1. 北京市医疗器械检验所体外诊断室
- Keywords:
isotope dilution liquid chromatography-tandem mass spectrometry (ID-LC/MS/MS);
5-methyltertrahydrofolate
- From:
Chinese Journal of Clinical Laboratory Science
2018;36(3):161-165
- CountryChina
- Language:Chinese
-
Abstract:
Objective To develop an isotope dilution liquid chromatography tandem mass spectrometry (ID-LC-MS/MS) method for determining the concentration of 5-methyltertrahydrofolate (5MT) in human serum.Methods After qualitative analysis of 5MT reference,its purity was identified by two methods with different principle,i.e.,mass balance and quantitative nuclear magnetic resonance.The definite value and uncertainty of 5MT concentrations in 6 serum samples containing same level of 5MT were assessed by ultra performance liquid chromatography-triple quadrupole tandem mass spectrometer equipped with electrospray ionization in monitoring mode of multiple reactions,in which 13C5-5MT was used as the internal standard.The precision,accuracy,limit of detection (LOD) and limit of quantification (LOQ) of the method were also evaluated.Results After qualitative analysis for confirming the principal component in the sample,the purity of 5MT reference was measured as 76.65％ by mass balance and quantitative nuclear magnetic resonance method.The definite value of 5MT in the serum sample was 8.22 ng/mL,the extended uncertainty was 0.54 ng/mL,LOD was 0.05 ng/mL,LOQ was 0.50 ng/mL and the repeatability and inter-batch precision CV were within 5.0％.The accuracy of measured results was verified by using NIST SRM 1955 since all the values of 3 levels were within the range of certified value with the extended uncertainty.Conclusion A accurately quantitative determination for the concentration of 5 MT in human serum with high specificity and sensitivity was established by using ID-LC-MS/MS method.
