Methylmalonic Acid Analysis using Urine Filter Paper Samples to Screen for Metabolic Vitamin B ₁₂ Deficiency in Older Adults

Abstract

Aim: Methylmalonic acid (MMA) analysis in urine represents a noninvasive approach to screening for vitamin B12 deficiency in older adults. A method allowing the analysis of MMA/creatinine in fasting urine collected on filter paper was developed/validated. Method: Dry urine specimens were eluted using a solution containing internal standards, filtrated and analyzed by ultra-performance LC-MS/MS. Results: The method allowed the chromatographic separation of MMA from succinic acid. Dried urine samples were stable for 86 days at room temperature. The MMA/creatinine ratios measured in urine collected on filter paper were highly correlated with values derived from the corresponding liquid specimens. Conclusion: This robust filter paper method might greatly improve the accessibility and cost–effectiveness of vitamin B12 deficiency screening in older adults.

A Rapid, Simple, Trace, Cost-Effective, and High-Throughput Stable Isotope-Dilution Liquid Chromatography–Tandem Mass Spectrometry Method for Serum Methylmalonic Acid Quantification and Its Clinical Applications

“…The analytical performances of the previous LC-MS/MS methods have been studied [ 14 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 ]. The LOD and LOQ of previous methods have ranged from 0.03–1.95 μmol/L and 0.03–4.20 μmol/L, respectively.…”

“…However, GC-MS-based MMA testing has several drawbacks (including high costs, cumbersome and laborious procedures, large sample volume, and long analytical time) [ 19 , 20 ]. Liquid chromatography–tandem mass spectroscopy (LC-MS/MS) methods are currently preferred for MMA measurements [ 14 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 ].…”

“…There are many published papers on MMA measurements by LC-MS/MS, using derivatized, non-derivatized, more or less complicated sample extraction, etc. Table 1 summarizes the reported analytical procedures of MMA measurement based on LC-MS/MS (from 2000 to 2022 [ 14 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 ]). Given a comprehensive consideration of all the method procedures as well as the aspects of cost-effectiveness, testing speeding, convenience, method performance, purpose, degree of difficulty for introducing the testing, robustness, accuracy, safety, and environmental protection, it is evident that all of these methods have their advantages and disadvantages.…”

“…Given a comprehensive consideration of all the method procedures as well as the aspects of cost-effectiveness, testing speeding, convenience, method performance, purpose, degree of difficulty for introducing the testing, robustness, accuracy, safety, and environmental protection, it is evident that all of these methods have their advantages and disadvantages. The improvement history of MMA testing shows that the method procedures basically consist of one or more of the following steps: multistep solid-phase extraction (SPE), derivations, protein precipitation, and ultrafiltration, while to complete the above step, at least one of the processes of evaporations, incubations, heating, dryings, reconsititutions, or centrifugations is required [ 14 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 ]. Improvements in procedures have usually focused on reducing these steps by changing different reagents or materials or by applying new materials [ 14 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 ].…”

“…The improvement history of MMA testing shows that the method procedures basically consist of one or more of the following steps: multistep solid-phase extraction (SPE), derivations, protein precipitation, and ultrafiltration, while to complete the above step, at least one of the processes of evaporations, incubations, heating, dryings, reconsititutions, or centrifugations is required [ 14 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 ]. Improvements in procedures have usually focused on reducing these steps by changing different reagents or materials or by applying new materials [ 14 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 ]. These improvements have reduced time and cost.…”