A Novel LC-MS/MS Assay for Vitamin B1, B2 and B6 Determination in Driedblood Spots and Its Application in Children. (29th May 2020)

• Record Type: Journal Article
• Authors: Wang, Fang; Zhang, Min; Huang, Xiaolan; Yang, Jian; Pei, Pei; Zhang, Ting; Cui, Xiaodai
• Is Part Of: Current developments in nutrition. Volume 4(2020)Supplement 2
• Page Start: 1849
• Page End: 1849
• Publication Date: 2020-05-29
• Subjects: Nutrition -- Periodicals; Nutritional Physiological Phenomena; Nutrition; Periodicals; Periodicals; Fulltext; Internet Resources; Periodicals; 612.3
• DOI: 10.1093/cdn/nzaa067_076
• Languages: English
• ISSNs: 2475-2991

Abstract

Abstract: Objectives: Several HPLC assays can be used to determine vitamin B1 (thiamin), B2 (riboflavin) and B6 (PLP) in biological matrix. However, no existing LC-MS/MS methods have been developed for quantifying thiamin, riboflavin and PLP in dried blood spots (DBSs), which are often used as a sampling/storage vessel for finger blood from infants and children. This study firstly used vacuum package in DBS cards storage, and evaluated the validity, reliability and stability of an LC-MS/MS assay for measuring thiamin, riboflavin and PLP in DBS cards. Methods: DBS samples were deproteinized by adding trichloroacetic acid containing thiamine-[13C4], riboflavin-[13C4, 15N2] and pyridoxal-d3 as internal standards. Thiamin, riboflavin and PLP were separated on a C8 column with a 5-min run time. Results: Thiamin, riboflavin and PLP were stable on the DBS card during at least 15 days of room-temperature storage under vacuum in the dark. Both the between-run and within-run variable coefficients (CV% values) were <8.56%. The accuracies (relative errors RE% values) were from −7.40% to 3.12%. Lower limits of quantifications (LLOQs) ranged from 0.2 to 0.5 ng/mL, and the recoveries were from 81.49% to 112.23% for all 3 analytes. Measurements of thiamin, riboflavin and PLP in the DBSs showed good agreement with the corresponding concentrations determined from liquid blood (R2 values >0.96). Conclusions: This validated method can be applied to the nutritional assessment of vitamins B1, B2, and B6. Funding Sources: This work was supported by the National Natural Science Foundation of China (Grant No. 81, 400, 848), Beijing Municipal