ArticlePloS one2015
Simultaneous Determination of Underivatized Vitamin B1 and B6 in Whole Blood by Reversed Phase Ultra High Performance Liquid Chromatography Tandem Mass Spectrometry.
Article in PloS one, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 43 citations in OpenAlex.
- Trial
- Seminal Homocysteine Levels in Men With Asthenozoospermia: Correlation With Sperm Parameters and Vitamin BTheScientificWorldJournal · 2026Article
- A Simultaneous Determination of the BInternational journal of molecular sciences · 2025Article
- Effect of Smoking on Seminal Plasma Vitamin BJournal of nutrition and metabolism · 2024Article
- A Novel Two-Dimensional Liquid Chromatography Combined with Ultraviolet Detection Method for Quantitative Determination of Pyridoxal 5'-Phosphate, 4-Pyridoxine Acid and Pyridoxal in Animal Plasma.Animals : an open access journal from MDPI · 2023Article
- Thiamine deficiency in pregnancy and lactation: implications and present perspectives.Frontiers in nutrition · 2023Review
- Men with oligozoospermia had lower level of seminal plasma pyridoxine compared to normozoospermic men.Heliyon · 2022Article
- Feasibility of Mass-Spectrometry to Lower Cost and Blood Volume Requirements for Assessment of B Vitamins in Patients Undergoing Bariatric Surgery.Metabolites · 2020Article
- Comprehensive and quantitative profiling of B vitamins and related compounds in the mammalian liver.Journal of chromatography. B, Analytical technologies in the biomedical and life sciences · 2020Article
- Supramolecular Tandem Assay for Pyridoxal-5'-phosphate by the Reporter Pair of Guanidinocalix[5]Arene and Fluorescein.ChemistryOpen · 2019Article
- Thiamine deficiency disorders: diagnosis, prevalence, and a roadmap for global control programs.Annals of the New York Academy of Sciences · 2018Article
- Pyridoxal 5'-phosphate assay based on lucigenin chemiluminescence.Mikrochimica acta · 2018Article
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Authors and funding
4 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundVitamin B1 (thiamine-diphosphate) and B6 (pyridoxal-5'phosphate) are micronutrients. Analysis of these micronutrients is important to diagnose potential deficiency which often occurs in elderly people due to malnutrition, in severe alcoholism and in gastrointestinal compromise due to bypass surgery or disease. Existing High Performance Liquid Chromatography (HPLC) based methods include the need for derivatization and long analysis time. We developed an Ultra High Performance Liquid Chromatography Tandem Mass spectrometry (UHPLC-MS/MS) assay with internal standards for simultaneous measurement of underivatized thiamine-diphosphate and pyridoxal-5'phosphate without use of ion pairing reagent.
methodsWhole blood, deproteinized with perchloric acid, containing deuterium labelled internal standards thiamine-diphosphate(thiazole-methyl-D3) and pyridoxal-5'phosphate(methyl-D3), was analyzed by UHPLC-MS/MS. The method was validated for imprecision, linearity, recovery and limit of quantification. Alternate (quantitative) method comparisons of the new versus currently used routine HPLC methods were established with Deming regression.
resultsThiamine-diphosphate and pyridoxal-5'phosphate were measured within 2.5 minutes instrumental run time. Limits of detection were 2.8 nmol/L and 7.8 nmol/L for thiamine-diphosphate and pyridoxal-5'phosphate respectively. Limit of quantification was 9.4 nmol/L for thiamine-diphosphate and 25.9 nmol/L for pyridoxal-5'phosphate. The total imprecision ranged from 3.5-7.7% for thiamine-diphosphate (44-157 nmol/L) and 6.0-10.4% for pyridoxal-5'phosphate (30-130 nmol/L). Extraction recoveries were 101-102% ± 2.5% (thiamine-diphosphate) and 98-100% ± 5% (pyridoxal-5'phosphate). Deming regression yielded slopes of 0.926 and 0.990 in patient samples (n = 282) and national proficiency testing samples (n = 12) respectively, intercepts of +3.5 and +3 for thiamine-diphosphate (n = 282 and n = 12) and slopes of 1.04 and 0.84, intercepts of -2.9 and +20 for pyridoxal-5'phosphate (n = 376 and n = 12).
conclusionThe described UHPLC-MS/MS method allows simultaneous determination of underivatized thiamine-diphosphate and pyridoxal-5'phosphate in whole blood without intensive sample preparation.
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