Establishment of a method for simultaneous detection of serum folate and metabolites,as well as methotrexate blood concentration
10.3969/j.issn.1673-4130.2024.22.001
- VernacularTitle:建立血清叶酸及代谢物和甲氨蝶呤血药浓度同时检测的方法
- Author:
Zhaoxin CHEN
1
;
Wenqi SONG
Author Information
1. 国家儿童医学中心/首都医科大学附属北京儿童医院检验中心,北京 100045
- Keywords:
liquid chromatography-tandem mass spectrometry;
folate;
methotrexate
- From:
International Journal of Laboratory Medicine
2024;45(22):2689-2693,2698
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish an analytical method based on ultra-high performance liquid chromatogra-phy-tandem mass spectrometry(UPLC-MS/MS)technology for simultaneous detection of serum folate and its metab-olites 5-methyltetrahydrofolate,formyl tetrahydrofolate,and methotrexate concentration.Methods Target an-alytes in samples were concentrated and enriched using solid-phase extraction,separated by an Acquity UPLC HSS T3 chromatographic column(2.1 mm×100 mm,1.8 μm),and eluted with a gradient using 0.1％formic acid water and acetonitrile as the mobile phase.The method was established using an ultra-high performance liquid chromatography-tandem quadrupole mass spectrometer(UPLC I-Class Xevo TQ-S micro IVD)and ac-cording to CLSI C62-A and other documents,the performance of this method in terms of quantification limit,linear range,precision,accuracy,dilution effect,carrying contamination,specificity,interference testing and so on was evaluated.Results The quantitation limit for folate was 1 ng/mL,and the quantitation limits for 5-methyltetrahydrofolate,formyltetrahydrofolate,and methotrexate were 5 ng/mL.Folate had a good linear re-lationship in the concentration range of 1-200 ng/mL(R2＞0.99),and 5-methyltetrahydrofolate,formyltet-rahydrofolate,and methotrexate had linear ranges of 5-2 000 ng/mL(R2＞0.99).The intra-day precision ranged from 1.7％to 13.9％,and the inter-day precision ranged from 2.7％to 14.6％.The spiked sample re-covery rates ranged from 93.6％to 108.9％,showing good precision and accuracy.Other examination results,such as dilution effect,carryover,and specificity,met evaluation requirements.Conclusion This study estab-lished an accurate and efficient UPLC-MS/MS method that allows for the simultaneous detection of serum fo-late and its metabolites and methotrexate blood concentrations.This method not only aids clinicians in simul-taneously monitoring methotrexate blood concentration and calcium formyl tetrahydrofolate concentration,but also enables dynamic observation of changes in total folate and active folate levels in patients during the treat-ment process.It lays the foundation for developing a reasonable rescue plan for calcium folinate and further improving the safety and effectiveness of methotrexate treatment.
