Vitamin Metabolomics

Vitamin metabolites are intermediates and end products formed as vitamins are absorbed, transformed, and degraded in vivo—including precursors, active metabolites, and degradation products such as the active vitamin D form 25-hydroxyvitamin D3, the folate metabolite 5-methyltetrahydrofolate, and the active vitamin B6 form pyridoxal phosphate. As a core component of the nutritional metabolic network, vitamin metabolites reflect vitamin absorption and utilization efficiency and participate in energy metabolism, gene expression regulation, and immune function. Their dysregulation is linked to chronic diseases (diabetes, cardiovascular disease), neurodegenerative disorders, and birth defects. Compared with single-vitamin assays, panoramic vitamin-metabolite analysis captures the full 'intake–conversion–utilization' pathway, providing deeper molecular evidence for nutritional assessment, early disease warning, and mechanistic research. It is a research hotspot in nutrition, clinical medicine, and translational medicine, applied in personalized nutrition, chronic-disease risk assessment, therapeutic drug monitoring, and special-population health management.

Principle Introduction

Vitamin-metabolite detection follows a 'panoramic extraction–efficient separation–precise qualitative/quantitative' workflow: 1. Extraction: mixed solvents (methanol–acetonitrile) or buffer co-extract fat- and water-soluble metabolites, with SPE removing protein/lipid impurities. 2. Separation: UHPLC separates metabolites by polarity, hydrophobicity, and molecular weight. 3. Identification & quantification: QQQ-MS or HRMS enable precise identification via characteristic precursor–product ion pairs, retention time, and library matching; stable-isotope internal standards (e.g., 25-hydroxyvitamin D3-d6, 5-methyltetrahydrofolate-d3) correct matrix effects and extraction loss for accurate quantification.

Literature

Published in Randomized Controlled Trial, 'Effect of Vitamin D and ω-3 Fatty Acid Supplementation on Risk of Age-Related Macular Degeneration: An Ancillary Study of the VITAL Randomized Clinical Trial' used vitamin-metabolite profiling to link vitamin D supplementation with age-related macular degeneration (AMD) risk. Subjects were randomized to vitamin D, ω-3, combined, or placebo groups over 5 years; serum vitamin D metabolites (25-hydroxyvitamin D3, 1,25-dihydroxyvitamin D3) were measured by UHPLC-QQQ-MS. Supplemented subjects showed significantly elevated 25-hydroxyvitamin D3 and 23% lower AMD risk (31% in those with baseline <20 ng/mL); combined supplementation offered no additional benefit over vitamin D alone. The study provides key metabolic evidence that vitamin D reduces AMD risk via its active metabolic pathway.

Sample Requirements

Serum / plasma

≥500 μL/sample, EDTA or heparin anticoagulated, -80°C sealed, dry-ice shipping; note fasting/postprandial time and supplement use.

Whole blood

≥1 mL/sample, EDTA anticoagulated, -80°C, dry-ice shipping; for folate, vitamin B12 metabolites; avoid hemolysis.

Urine

≥2 mL/sample, -80°C, dry-ice shipping; 0.1% sodium azide optional; note type (24 h / morning).

Tissue (animal / plant)

animal ≥1 g, plant ≥2 g, snap-frozen -80°C, dry-ice shipping; specify region and conditions.

Food / feed

≥5 g/sample, sealed -20°C, ice-pack shipping; specify type and processing state.

Deliverables

Sample QC report

integrity, extraction/internal-standard recovery, precision (RSD≤10%), stability.

Raw data

UHPLC-MS chromatograms, retention times, ion peaks, peak areas, library matching.

Data report

qualitative identification of 30+ vitamin metabolites, quantification table (ng/mL, μg/g), differential volcano plots, pathway enrichment, comparison with reference ranges, nutritional status assessment.

Technical support

pretreatment optimization, UHPLC-MS parameters, pathway guide, personalized nutrition recommendations.

FAQ

① very low endogenous concentration; ② improper pretreatment (temperature, solvent ratio); ③ poor storage (freeze-thaw, light-sensitive lipids); ④ suboptimal MS parameters. Optimize and re-run.

① add SPE/liquid-liquid extraction; ② adjust chromatography; ③ use MRM targeted scanning; ④ high-purity reagents and IS.

① inconsistent diet/sampling/dose; ② inconsistent handling; ③ instrument drift; ④ metabolic fluctuation. Standardize and calibrate; include replicates (RSD≤10%).

① spike recovery 85%–115%; ② multiple isotope IS; ③ certified reference material (CRM); ④ blanks; ⑤ replicate RSD≤10%.

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