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Abstract
Bariatric surgery induces deficiencies in a combination of B vitamins. However, high costs and a large blood volume requirement are barriers to routine screening. We adapted and validated a method coupling tandem mass spectrometry (MS/MS) with high-performance liquid chromatography (HPLC) to facilitate cost-effective analysis for simultaneous detection of B vitamins in low volumes of plasma. Based on existing methods, pooled plasma was extracted using hexane and acetonitrile and seven B vitamin analytes were separated using HPLC. Detection was performed with an Agilent 6460 triple quadrupole tandem mass spectrometer (MS/MS) using electrospray ionization in the positive ion mode. We evaluated linearity, recovery, precision, and limit of detection, as well as costs of the assay. We evaluated seven B vitamins from plasma; five (riboflavin, nicotinamide, pantothenic acid, pyridoxine, and biotin) were detected and quantified with precision and linearity. Recovery ranged from 63 to 81% for each of the vitamins, except for nicotinamide—the recovery of which was suppressed to 40%, due to plasma matrix effects. We demonstrated the feasibility of the HPLC–MS/MS method for use in patients who undergo bariatric surgery by analyzing pooled plasma from patients with a lower cost and blood volume than had we sent the samples to a commercial laboratory. It is advantageous and feasible, in terms of low cost and blood volume requirement, to simultaneously measure plasma concentrations of B vitamins using HPLC–MS/MS. With further improvements, the method may enable personalized nutritional assessment for the nutritionally compromised, bariatric surgery population.
Highlights
B vitamins include thiamine, riboflavin, niacin, pantothenic acid, pyridoxine, biotin, folate and cobalamin, which are essential for humans
To assess the validity of our high-performance liquid chromatography (HPLC)–MS/MS methodology for simultaneous assessment of various B vitamins, we evaluated linearity, limit of detection, recovery, precision, extraction efficiency, and matrix effects
Linearity assessed by R2 found values for riboflavin, biotin, pantothenic acid, nicotinamide, and pyridoxine that were greater than 0.9950, indicating that the regression curves explained most the variability in the Metabolites 2020, 10, x FOR PEER REVIEW
Summary
B vitamins include thiamine (vitamin B1 ), riboflavin (vitamin B2 ), niacin (vitamin B3 ), pantothenic acid (vitamin B5 ), pyridoxine (vitamin B6 ), biotin (vitamin B7 ), folate (vitamin B9 ) and cobalamin (vitamin B12 ), which are essential for humans They are needed in energy and other basic metabolic pathways and red blood cell synthesis, and their deficiency comes with consequences such as anemia and poor nervous system function [1]. B vitamins are quantified using microbiological, immunological or enzymatic assays These require the separate quantification of each B vitamin, requiring a long time for analysis, as well as high costs and large sample volumes. Recent efforts to Metabolites 2020, 10, 240; doi:10.3390/metabo10060240 www.mdpi.com/journal/metabolites
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