Precision Quantification of Vitamin Metabolism Using Targeted LC-MS/MS Analysis
Accurate vitamin analysis is essential for nutritional research, food safety, and pharmaceutical applications. Leveraging liquid chromatography-tandem mass spectrometry (LC-MS/MS) with isotope-labeled internal standards, we offer an ultra-sensitive and high-precision quantification service for over 30 water- and fat-soluble vitamins.
By overcoming the limitations of conventional methods, our vitamin-targeted metabolomics platform provides researchers with unparalleled accuracy and depth in metabolic analysis.
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- Methods
- Metabolist Coverage
- Advantages
- Platform
- Workflow
- Sample Requirement
- Report
- FAQ
- Publication
Methods for Vitamin Analysis
Vitamins play a crucial role in metabolic regulation, influencing disease progression, nutritional interventions, aging, and immune function. As research into vitamin metabolism advances, the demand for accurate quantification continues to grow. However, traditional detection methods such as HPLC and ELISA present significant limitations:
Limited Sensitivity: Struggle to detect trace levels of vitamins, including serum vitamin D3 and K2.
Low Specificity: Inability to distinguish structurally similar isomers, such as α- and γ-tocopherol (vitamin E).
Low Throughput: Conventional assays analyze only a few markers per experiment, making it challenging to map complex metabolic networks.
Several analytical methods are available, each with unique advantages and limitations. Below is a comparative overview of four common techniques:
| Method | Advantages | Limitations | Suitable for |
|---|---|---|---|
| UV-Vis Spectrophotometry | Cost-effective, easy to use | Prone to interference, lengthy process | Vitamin A, Vitamin D |
| Fluorescence Spectrophotometry | High sensitivity for fluorescent compounds | Limited to naturally fluorescent vitamins | Vitamin B1, B2, C |
| Gas Chromatography (GC) | Fast, highly sensitive | Poor reproducibility | Vitamin E |
| High-Performance Liquid Chromatography (HPLC) | High sensitivity, low detection limits, robust for complex samples | Higher cost | Most water- and fat-soluble vitamins |
Why HPLC is the Gold Standard in Vitamin Analysis
Among these methods, high-performance liquid chromatography (HPLC) has become the preferred technique for vitamin quantification. It offers:
High Sensitivity & Accuracy – Capable of detecting minute vitamin concentrations, even in complex biological matrices.
Versatile Applications – Suitable for both water-soluble (e.g., vitamin C, B-complex) and fat-soluble vitamins (e.g., vitamins A, D, E, K).
Structural Differentiation – Efficiently separates vitamin isomers, providing precise quantification of individual compounds.
Broad Industry Adoption – Widely used in pharmaceuticals, clinical diagnostics, and food quality testing due to its reliability.
Sample Requirements & Testing Parameters
HPLC-based vitamin analysis requires specific sample volumes depending on the vitamin type:
| Vitamin Category | Vitamin Type | Detection Method | Sample Requirement |
|---|---|---|---|
| Water-Soluble Vitamins | Vitamin C | HPLC | 1–2 g / 1–2 mL |
| Vitamin B1, B2, B3, B6, B9, B12 | HPLC | 1–2 g / 1–2 mL | |
| Fat-Soluble Vitamins | Vitamin A, D3, E | HPLC | 1–2 g / 1–2 mL |
With HPLC's superior precision and broad applicability, it remains the go-to method for vitamin detection, ensuring accurate and reproducible results across diverse sample types.
Complete Coverage: Water-Soluble & Fat-Soluble Vitamins
Understanding vitamin metabolism is essential for research in nutrition, disease mechanisms, and metabolic regulation. Our LC-MS/MS-based vitamin analysis service offers precise quantification of both water-soluble and fat-soluble vitamins in various biological and food samples, ensuring high sensitivity, accuracy, and reproducibility.
| Category | Vitamin Type | Compounds |
|---|---|---|
| Water-Soluble Vitamins | Vitamin B Complex | B1 (Thiamine), B2 (Riboflavin), B3 (Niacin), B5 (Pantothenic acid), B6 (Pyridoxine), B7 (Biotin), B9 (Folate), B12 (Cobalamin) |
| Vitamin C | Ascorbic acid, Dehydroascorbic acid (oxidized form) | |
| Fat-Soluble Vitamins | Vitamin A | Retinol, Beta-carotene, Retinoic acid |
| Vitamin D | 25(OH)D2, 25(OH)D3, 1,25(OH)₂D3 (active metabolite) | |
| Vitamin E | α/β/γ/δ-Tocopherols, Tocotrienols | |
| Vitamin K | K1 (Phylloquinone), MK-4 to MK-13 (Menaquinones) |
Technical parameters:
- Ultra-High Sensitivity: Detects as low as 0.1 pg/mL (e.g., Vitamin D3), suitable for micro-volume samples (10 μL serum).
- Exceptional Accuracy: Isotope-labeled internal standards ensure >90% recovery, with intra- and inter-batch CVs<5%.
- Broad Sample Compatibility: Supports serum, plasma, tissues, cells, food, and plant extracts.
Cutting-Edge Platform: Thermo Q Exactive UPLC-MS/MS system (≥70,000 FWHM resolution) for high-throughput, high-precision analysis.
Why Choose Our Service?
- Leading-Edge Technology for Reliable Results
High-Resolution LC-MS/MS: Sciex Triple Quad™ 6500+ Mass Spectrometer & Agilent 1290 Infinity II UPLC deliver superior sensitivity and reproducibility.
- Rigorous Quality Control
Optimized Sample Processing: Proprietary solid-phase extraction (SPE) eliminates phospholipid interference, improving data integrity.
Comprehensive Data Reporting: Includes raw data, concentration values, pathway enrichment analysis, and publication-ready visualizations.
- All-in-One Detection for Maximum Efficiency
Simultaneous Analysis: Measures both water-soluble and fat-soluble vitamins in a single assay, reducing batch-to-batch variability and costs.
Metabolite Co-Analysis: Detects key vitamin derivatives (e.g., FAD, 5-MTHF) to identify metabolic pathway disruptions.
- Advanced Quantification Strategies
Stable Isotope Labeling: 13C/15N-labeled internal standards ensure quantification accuracy within ±10%.
Matrix-Matched Calibration: Uses real biological matrices for standard curve generation, eliminating matrix effects.
- In-Depth Data Mining & Biomarker Discovery
Multi-Omics Integration: Optional analysis with lipidomics and metabolomics to reveal vitamin-related metabolic interactions.
Biomarker Identification: Uses OPLS-DA, ROC curve analysis for differential metabolite screening and validation.
Detection Platform
Agilent 1290-6470 (image source: agilent.com)
Sciex QTRAP 6500+ (image source: Sciex QTRAP 6500+)
Streamlined Service Workflow: Fast & Reliable Delivery
Comprehensive Data Outputs
Basic Analysis: Raw data, concentration tables, and quality control reports.
Advanced Insights: PCA, hierarchical clustering, OPLS-DA, KEGG/Reactome pathway enrichment.
Visual Reports: Volcano plots, heatmaps, metabolic network diagrams, publication-ready figures (Word/PDF format).
With state-of-the-art technology, rigorous validation, and expert support, our targeted vitamin metabolomics service empowers researchers with unparalleled accuracy and depth in metabolic profiling.
Sample Requirements
| Sample type | Recommended sample size |
|---|---|
| Tissue | >200 μL |
| Urine | 200-500 μL |
| Serum/plasma | >100 μL |
| Cerebrospinal fluid, amniotic fluid, bile and other body fluids | >200 μL |
| Suspension cells | >1*107 |
| Walled cells | >1*107 |
| Cell supernatant | >1 mL |
Report
- A detailed technical report will be provided at the end of the whole project, including the experiment procedure, MS/MS instrument parameters
- Analytes are reported as uM, while CV's are generally 10%.
- The name of the analytes, abbreviation, formula, molecular weight and CAS# would also be included in the report.
Comprehensive Research Support
Customizable Assays: Expand analysis to include amino acids, fatty acids, and other metabolites for network-based insights.
Expert Consultation: Personalized support on study design, statistical analysis, and manuscript-ready data visualization.
Preliminary Feasibility Assessment: Free sample size evaluation and collection strategy recommendations.
Data Interpretation Assistance: One-on-one consultations with metabolomics experts for optimized experimental planning.
Frequently Asked Questions (FAQ): Vitamin Analysis Services
Are detection methods consistent across different sample types (e.g., serum vs. tissue)?
No. Sample preparation varies based on the sample type. For instance, tissue samples require homogenization before extracting fat-soluble vitamins. We provide optimized standard operating procedures (SOPs) tailored to each sample type.
Can you measure the oxidized and reduced forms of vitamins (e.g., Vitamin C)?
Yes! We offer simultaneous quantification of both ascorbic acid (reduced form) and dehydroascorbic acid (oxidized form). To preserve oxidation-sensitive compounds, we recommend adding stabilizers during sample collection.
What analytical insights are included in the data report?
Beyond concentration measurements, our reports feature:
Metabolic pathway mapping to visualize vitamin-related biochemical interactions.
Differential metabolite screening (VIP >1, p < 0.05) to highlight statistically significant variations.
Correlation heatmaps for advanced data interpretation.
How should samples be stored after collection?
Serum/plasma: Centrifuge immediately (4°C, 3000 rpm, 10 min), aliquot, and store at -80°C.
Urine: Add antioxidants (e.g., ascorbic acid) before freezing at -80°C to prevent degradation. Avoid repeated freeze-thaw cycles.
Do you support special sample types?
Yes! We accommodate cell culture media, plant extracts, and food matrices. Please contact us in advance to discuss customized preprocessing protocols for your specific sample type.
Learn about other Q&A about other technologies.
Our Vitamins Analysis Review
- THE ROLE OF INTRA-TUMORAL MICROBIOME IN MODULATING MUCOSAL-ASSOCIATED INVARIANT T CELL (MAIT) FUNCTION IN LUNG CANCER (Doctoral dissertation, Johns Hopkins University). Yang, X. 2024. https://jscholarship.library.jhu.edu/items/f7b260ea-038e-4719-90e1-0da47a14711c
- Nutritional analysis of commercially available, complete plant-and meat-based dry dog foods in the UK. Brociek, R. A., Li, D., Broughton, R., & Gardner, D. S. 2024. https://doi.org/10.1101/2024.09.11.612409
- Fatty Acid and Antioxidant Profile of Eggs from Pasture-Raised Hens Fed a Corn- and Soy-Free Diet and Supplemented with Grass-Fed Beef Suet and Liver. Sergin, S., Jambunathan, V., et al. 2022. https://doi.org/10.3390/foods11213404
How to place an order:
*If your organization requires signing of a confidentiality agreement, please contact us by email
With integrated set of separation, characterization, identification and quantification systems featured with excellent robustness & reproducibility, high and ultra-sensitivity, Creative Proteomics provides reliable, rapid and cost-effective fat-soluble vitamins targeted metabolomics services.
