Cofactors and Vitamins Analysis Service

Why Cofactors and Vitamin-Derived Metabolites Matter

Many vitamins become biologically active only after conversion into cofactor forms. These forms participate in core processes such as redox control, mitochondrial energy production, and biosynthetic regulation.

In pathway-focused research, changes in cofactor pools or redox ratios can reveal enzyme constraints and metabolic bottlenecks. These signals may be missed when relying on broad, untargeted metabolomics alone.

Targeted measurement helps reduce ambiguity and supports more direct mechanistic interpretation.

Research Questions This Service Helps Address

This analysis is commonly used to support questions such as:

• Are enzyme-dependent pathways limited by cofactor availability?
• Is cellular redox balance shifted under disease or treatment conditions?
• Do nutrition or intervention strategies change vitamin-dependent metabolism?
• Which pathway nodes show the strongest functional disruption?

Service Overview: Vitamins and Cofactors Profiling

Our platform offers modular analytical suites designed to match specific research intents. Each portfolio is optimized for absolute quantification using validated isotope dilution methods.

Redox & Energy Homeostasis

Designed for studies on mitochondrial function and cellular "fuel" states.

• NAD+/NADH & NADP+/NADPH
• FAD / FMN
• ATP / ADP / AMP
• Coenzyme A (CoA) derivatives

One-Carbon & Epigenetics

Focused on methyl donor availability and the Folate-B₁₂ axis.

• 5-MTHF & Active Folates
• SAM / SAH Flux
• Vitamin B12 (Cobalamins)
• B6 Vitamers (PLP/PMP)

Comprehensive Profiling

Wide-spectrum screening for unbiased discovery and nutritional research.

• 50+ Essential Vitamins
• Bioactive Metabolites
• Fat-soluble (A/D/E/K)
• Water-soluble (B/C)

Custom Pathway Panel

Flexible integration of specific precursors and downstream products.

• Tailorable Analyte List
• Matrix-Specific Optimization
• Pathway-Focused Validation
• Multi-Omics Integration

Comprehensive Analyte Coverage: 50+ Vitamins and Coenzymes

Below is a consolidated list that merges the earlier table with your expanded target set. Final panel composition can be adapted to the study question, sample matrix, and analyte chemistry.

Note: Some lipid-soluble vitamins and hydrophobic cofactors may require dedicated extraction and LC conditions. Final target coverage is confirmed during panel design based on sample type and study goals.

Advantages of Our Analysis Platform

• Unrivaled Sensitivity: Detection limits reach the low picogram (pg/mL) level, enabling high-fidelity profiling in limited or rare samples such as biopsies, primary cells, and exosomes.
• Exceptional Quantitative Linearity: We achieve R2 > 0.99 linearity across a 4-6 order of magnitude dynamic range.
• Exceptional Reproducibility: Intraday and interday CV (Coefficient of Variation) < 15%, ensuring reliable data for longitudinal studies.
• Advanced Isotope Dilution: We utilize 30+ stable isotope-labeled internal standards to correct for matrix effects and extraction losses.

Workflow for Cofactors and Vitamins Analysis

Analytical Platforms for Vitamins and Cofactor Analysis

We utilize industry-leading LC-MS/MS and Orbitrap technologies to deliver sub-picogram sensitivity and superior isomeric resolution for complex biological matrices.

Platform Core Advantages:

• Isomeric Resolution: Superior separation of complex B-vitamin vitamers and cofactor derivatives.
• Matrix Mitigation: Advanced iFunnel technology and isotope dilution to minimize ion suppression.
• Data Integrity: High-speed MRM monitoring and HRAM confirmation ensure zero-compromise accuracy.

Agilent 6495C Triple Quadrupole (Figure from Agilent)

Orbitrap Exploris 480 (Figure from Thermo)

Waters ACQUITY UPLC System (Figure from Waters)

Sample Submission & Preparation Guidelines

To ensure the highest data integrity, samples must be flash-frozen immediately upon collection to preserve labile cofactors and vitamins. Please refer to the following requirements for different sample matrices:

Important Technical Notes for Cofactor Analysis:

• Flash Freezing: For NAD(H), NADP(H), and Coenzyme A derivatives, immediate freezing in liquid nitrogen is mandatory to prevent enzymatic interconversion.
• Freeze-Thaw Cycles: Strictly avoid repeated freeze-thaw cycles, as this leads to the rapid degradation of B-group vitamins and unstable coenzymes.
• Light Sensitivity: Vitamins A, B12, and Folate are highly light-sensitive. Please use light-protected (amber) tubes or wrap samples in aluminum foil if possible.
• Custom Matrices: For unique matrices (e.g., milk, exosomes, or culture media), please contact our technical team for specialized extraction protocols.

Comprehensive Deliverables: Accurate Data for Your Research

• Standardized Raw Data: Full access to raw LC-MS files (.raw or .d).
• Quantification Report: An Excel/CSV table containing absolute concentrations (nmol/g or ng/mL) for all analytes.
• Methodology Section: A detailed description of the SOPs, instrument parameters, and chemicals used (ready for your "Materials and Methods" section).
• Statistical Analysis: PCA, Heatmaps, and Fold-Change analysis between experimental groups.
• Pathway Mapping: High-resolution visualization of cofactor changes within central carbon metabolism.

UPLC-MS/MS MRM data showing baseline separation of B6 isomers (e.g., PLP), validating superior resolution for active forms.

Standard curve (R²=0.9998) spanning 10 pg/mL to 10 μg/mL. Inset highlights high sensitivity and precision at the LLOQ.

Heatmap of 50 cofactors distinguishing groups. Asterisks indicate significant (p<0.05) shifts in key metabolites like NAD+.

Integrated pathway map showing B-vitamin drive. Node color indicates concentration; red highlights key enzymatic bottlenecks.

Research Applications for Vitamins and Cofactor Analysis

Agricultural Biofortification & Stress Resilience

Quantifying vitamin-to-cofactor conversion rates to enhance the nutritional density of biofortified crops and improve plant resistance to abiotic stresses (e.g., drought, salinity).

Metabolic Engineering & Synthetic Biology

Monitoring NAD(P)H and CoA availability in microbial cell factories to optimize metabolic flux for high-yield biosynthesis of high-value chemicals and biofuels.

Industrial Fermentation & Bioprocess Optimization

Tracking real-time vitamin depletion and coenzyme turnover in bioreactors to refine media formulations, prevent metabolic bottlenecks, and maximize product consistency.

Functional Food Stability & Bioaccessibility

Assessing the thermal and oxidative stability of bioactive vitamins in fortified foods through simulated digestion studies to determine true bioavailability and shelf-life.

Environmental Ecotoxicology & Biomarker Discovery

Evaluating how environmental pollutants disrupt mitochondrial energy cofactors, utilizing changes in redox-active species as sensitive bioindicators of ecosystem health.

Biopharmaceutical Cell Culture Development

Ensuring the metabolic vigor and consistency of mammalian cell lines (e.g., CHO cells) in large-scale production by monitoring essential cofactor pools required for therapeutic protein synthesis.