海南医科大学学报2025，Vol.31Issue(15)：1163-1171,9.DOI:10.13210/j.cnki.jhmu.20241122.001
基于替代分析物的LC-MS/MS检测儿童末梢血中25-羟维生素D方法建立及评价
Establishment and evaluation of an LC-MS/MS method for the detection of 25-Hydroxyvitamin D in children's capillary blood using surrogate analytes
摘要
Abstract
Objective:To establish a liquid chromatography-tandem mass spectrometry(LC-MS/MS)method for the detec-tion of 25-hydroxyvitamin D in children's capillary blood using isotopic surrogate analytes.Methods:Isotope-labeled substances 25(OH)D2-d6 and 25(OH)D3-d3 were used as surrogate analytes and 25(OH)D3-d6 was used as the internal standard.After pro-tein precipitation treatment,the LC-MS/MS method was applied for detection and the contents of 25(OH)D2 and 25(OH)D3 in actual specimens were obtained through formula conversion.The method was evaluated for the response value equivalency at the same concentration,specificity,linearity,limit of detection(LOD),limit of quantification(LOQ),accuracy,precision,matrix effect,carryover contamination and recovery of spiked actual samples.Results:The response factors for 25(OH)D2 and 25(OH)D3 were 1.46 and 1.04,respectively;the linearity correlation coefficients were all>0.995,with LODs of 0.16 and 0.33 ng/mL,and LOQs of 0.5 and 1 ng/mL,respectively.The relative recoveries of the isotopic surrogate analytes 25(OH)D2-d6 and 25(OH)D3-d3 were within the range of 96.05%-107.07%,with intra-batch/inter-batch precisions all less than 10%,matrix factors be-tween 0.882-1.121,There was no obvious carry-over contamination.The samples could be stored at room temperature away from light for 24 hours,or at 2-8℃ for 7 days.Conclusion:A method for the quantitative determination of 25(OH)D2 and 25(OH)D3 in children's capillary blood using LC-MS/MS with isotopic surrogate analytes has been established.This method is straightfor-ward,practical,and reliable,and is anticipated to be a candidate method for children or other populations whose blood collection is challenging.关键词
液相色谱法/串联质谱法/25-羟维生素D/末梢血/替代分析物Key words
Liquid chromatography/Tandem mass spectrometry/25-Hydroxyvitamin D/Capillary blood/Isotopic Surro-gate analytes分类
医药卫生引用本文复制引用
刘菲,宋晶晶,曹明鹏,祖琴琴,周曙光,张爱喜,王丽,李明..基于替代分析物的LC-MS/MS检测儿童末梢血中25-羟维生素D方法建立及评价[J].海南医科大学学报,2025,31(15):1163-1171,9.基金项目
This study was supported by Open Project of the Key Laboratory of Birth Population Health Education and Anhui Maternal and Child Health Association(JK202213) （JK202213）
Anhui Medical University Affiliated Hospital Youth Science and Technology Talent Training Project"Eagle Plan"(2023cy010) 出生人口健康教育部重点实验室联合安徽省妇幼保健协会开放课题(JK202213) （2023cy010）
安徽医科大学附属医院青年科技人才培养项目"雏鹰计划"(2023cy010) （2023cy010）
