An LC-MS/MS method for serum methylmalonic acid suitable for monitoring vitamin B12 status in population surveys

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• 作者：Ekaterina M. Mineva ; Mindy Zhang…
• 关键词：MMA ; Method comparison ; GC/MS ; Anticoagulant types ; NHANES ; NIST SRM 1950 ; Vitamin B12
• 刊名：Analytical and Bioanalytical Chemistry
• 出版年：2015
• 出版时间：April 2015
• 年：2015
• 卷：407
• 期：11
• 页码：2955-2964
• 全文大小：546 KB
• 参考文献：Dietary Reference Intakes: Thiamin, riboflavin, niacin, vitamin B6, folate, vitamin B12, pantothenic acid, biotin, and choline. National Academy Press, Washington, DC
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• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Analytical Chemistry
  Food Science
  Inorganic Chemistry
  Physical Chemistry
  Monitoring, Environmental Analysis and Environmental Ecotoxicology
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1618-2650

文摘

Methylmalonic acid (MMA), a functional indicator of vitamin B12 insufficiency, was measured in the US population in the National Health and Nutrition Examination Survey (NHANES) from 1999 to 2004 using a GC/MS procedure that required 275?μL of sample and had a low throughput (36 samples/run). Our objective was to introduce a more efficient yet highly accurate LC-MS/MS method for NHANES 2011-014. We adapted the sample preparation with some modifications from a published isotope-dilution LC-MS/MS procedure. The procedure utilized liquid–liquid extraction and generation of MMA dibutyl ester. Reversed-phase chromatography with isocratic elution allowed baseline resolution of MMA from its naturally occurring structural isomer succinic acid within 4.5?min. Our new method afforded an increased throughput (?60 samples/run) and measured serum MMA with high sensitivity (LOD--2.1?nmol/L) in only 75?μL of sample. Mean (±SD) recovery of MMA spiked into serum (2 d, 4 levels, 2 replicates each) was 94?%?±-.5?%. Total imprecision (41 d, 2 replicates each) for three serum quality control pools was 4.9?%-.9?% (97.1-48?nmol/L). The LC-MS/MS method showed excellent correlation (n--26, r--.99) and no bias (Deming regression, Bland-Altman analysis) compared to the previous GC/MS method. Both methods produced virtually identical mean (±SD) MMA concentrations [LC-MS/MS: 18.47?±-.71?ng/mL (n--7), GC/MS: 18.18?±-.67?ng/mL (n--1)] on a future plasma reference material compared with a GC/MS method procedure from the National Institute of Standards and Technology [18.41?±-.70?ng/mL (n--5)]. No adjustment will be necessary to compare previous (1999-004) to future (2011-014) NHANES MMA data. Graphical Abstract ?/em>