分析化学2024，Vol.52Issue(3)：373-379,7.DOI:10.19756/j.issn.0253-3820.221316
基于非衍生化串联质谱技术分析新生儿滤纸干血片中的乳清酸
Analysis of Orotic Acid in Dry Blood Spots from Newborns by Non-derivatized Flow Injection Analysis Coupled to Tandem Mass Spectrometry
摘要
Abstract
Orotic acid(ORA)is a key targeted biomarker in inherited metabolic diseases,especifically ornithine transcarboxylase deficiency.However,ORA is not included into the current first-tier newborn screening program due to the absence of a suitable assay method.Therefore,some false positives and error results need to be corrected by a second-tier assay,which requires additional time and urine of newborns.The aim of this study was to develop a convenient method for analysis of ORA in dry blood spots(DBS)by non-derivatized flow injection analysis coupled to tandem mass spectrometry.ORA was extracted from DBS with 78%methanol aqueous solution containing ORA-15N2 as internal standard.The samples were analyzed in MRM mode with negative-mode ionization from a single injection in 2 min acquisition time.A linear response range was found in the ORA concentration range from 0.25 to 40.0 μmol/L,and the correlation coefficient was good(R2>0.99).The limit of detection of ORA(S/N=3)was 0.12 μmol/L.Matrix effects of the method was carefully evaluated and determined,and the difference of their response values was within 20%of the target,indicating that the matrix effects would not affect the accurate quantification of ORA.Then,the proposed method was successfully applied to analyze ORA in actual DBS samples.The recoveries of ORA were in the range of 101.4%-122.2%,and the relative standard deviation(RSD)was 0.8%-13.3%.Meanwhile,the residual contamination of the method was evaluated,and the results showed that the residual contamination was acceptable.Based on the method established in this study,900 clinical DBS samples from healthy newborns was analyzed.Herein,the lower limit cut-offs were set at below 1.0%,and the upper limit cut-offs were set at above 99.0%.Thus,the reference intervals of ORA(1.10-1.52 μmol/L)in DBS of this laboratory was preliminarily obtained,which provided a preliminary foundation for the promotion of NBS program in China in the near future.关键词
非衍生化流动注射串联质谱/乳清酸/滤纸干血片/新生儿/遗传代谢疾病/筛查Key words
Non-derivatized flow injection analysis coupled to tandem mass spectrometry/Orotic acid/Dry blood spot/Newborn/Inherited metabolic diseases/Screening引用本文复制引用
张潇分,樊静静,陈武炼,王燕敏,纪伟,田国力..基于非衍生化串联质谱技术分析新生儿滤纸干血片中的乳清酸[J].分析化学,2024,52(3):373-379,7.基金项目
国家自然科学基金项目(No.21803009)、上海市科学技术委员会科研项目(No.18441905100)、上海交通大学"交大之星"医工交叉项目(No.YG2022QN097)和上海市儿童医院院级课题国家自然科学基金培育专项项目(No.2021YGZQ09)资助. Supported by the National Natural Science Foundation of China(No.21803009),the Project of Shanghai Science Committee(No.18441905100),the Interdisciplinary Program of Shanghai Jiao Tong University(No.YG2022QN097)and the National Natural Science Foundation of China,a Hospital Level Project of Shanghai Children's Hospital(No.2021YGZQ09). （No.21803009）
