Nicotinate and Nicotinamide LC–MS/MS Analysis Service

Why Analyze Nicotinate and Nicotinamide?

Nicotinate and nicotinamide are central building blocks for the nicotinamide adenine dinucleotide pool. Through de novo synthesis from tryptophan and salvage pathways that recycle nicotinamide and nicotinamide riboside, cells maintain NAD and NADP levels to support glycolysis, the TCA cycle, oxidative phosphorylation, fatty acid oxidation, and many other redox-dependent reactions.

When precursor levels are perturbed, you may see:

• Imbalanced NAD⁺/NADH or NADP⁺/NADPH ratios.
• Compensatory changes in de novo versus salvage pathways.
• Accumulation of downstream catabolites such as trigonelline or nicotinurate.

By profiling nicotinate, nicotinamide, and their derivatives alongside broader metabolomics data, you can:

• Clarify whether changes in NAD metabolism are driven by precursor limitation, pathway bottlenecks, or increased consumption.
• Link redox changes to nutrient status, cofactor metabolism, and one-carbon pathways.
• Support mechanism-of-action studies for compounds that modulate NAD biosynthesis or salvage.

What Our Nicotinate and Nicotinamide Analysis Service Includes

Our nicotinate and nicotinamide analysis is built as a targeted metabolomics panel that can stand alone or be integrated into wider cofactor and vitamin profiling.

Typical service modules include:

• Absolute quantification of nicotinate and nicotinamide in biofluids, tissues, cells, and other research samples.
• Extended panel of related metabolites, including key intermediates and catabolites for pathway interpretation.
• Custom panel design to add or remove analytes that are relevant to your model or intervention.
• Optional integration with NAD species (NAD⁺, NADH, NADP⁺, NADPH) and one-carbon metabolites via linked services.
• Data analysis and visualization, including group comparisons, ratios (for example nicotinate/nicotinamide), and basic pathway mapping.

Targeted Metabolite Panel: Nicotinate and Nicotinamide Pathway

Advantages of Our Nicotinate and Nicotinamide Analysis Service

• Sensitive LC–MS/MS quantification: Targeted methods capture low-abundance nicotinate/nicotinamide in small-volume samples.
• High calibration linearity: Calibration curves are tuned for strong linearity, typically with R² ≥ 0.99 across working ranges.
• QC-anchored reproducibility: Isotope-labeled internal standards and pooled QCs help keep batch-to-batch drift under tight control.
• Easy extension to cofactor networks: The same platform can be expanded to include NAD species, vitamins, one-carbon or CoA-related metabolites when you need broader pathway coverage.

Analytical Platform for Nicotinate and Nicotinamide Analysis

Core Instruments

Our nicotinate and nicotinamide panel is run on targeted LC–MS/MS platforms commonly used in small-molecule metabolomics.

UHPLC systems: High-performance UHPLC with binary pumps and autosamplers for low-volume injections and stable gradients.

Triple quadrupole mass spectrometers (primary quantification)

Representative platforms used in targeted metabolomics, such as:

• Thermo Scientific TSQ series triple quadrupole systems.
• AB Sciex QTRAP or API series triple quadrupole instruments.

These are operated in electrospray ionization (ESI), typically in positive mode, using MRM for nicotinate, nicotinamide, and related metabolites.

High-resolution MS (method development and confirmation, when needed)

Orbitrap or TOF-based instruments (for example Thermo Q Exactive or similar HRMS systems) can be used to confirm identities and refine transitions before locking the routine LC–MS/MS method.

Method Performance Parameters

Triple Quad™ 6500+ (Figure from Sciex)

TSQ Altis Triple Quadrupole MS (Figure from Thermo Scientific)

Thermo Fisher Q Exactive (Figure from Thermo Fisher)

Agilent 1260 Infinity II HPLC (Fig from Agilent)

From Sample to Data: Nicotinate and Nicotinamide Analysis Workflow

1

Project consultation and panel design

Clarify your research questions, sample types, target analytes, and any co-panels (for example NAD, one-carbon metabolites).

2

Sample receipt and pre-analytical QC

Verify sample identity, volume or mass, container type, storage conditions, and freeze–thaw history; document any deviations.

3

Metabolite extraction and preparation

Perform protein precipitation and extraction under cold conditions, add internal standards, and, where needed, adjust pH or use stabilizing additives to protect labile metabolites.

4

LC–MS/MS acquisition

Separate pyridine metabolites under reversed-phase or mixed-mode chromatography and detect them by MRM in positive ion mode.

5

Data processing and quality control

Integrate peaks, review retention times, check calibration curves and QC samples, and flag any outliers or issues.

6

Quantitative reporting and optional interpretation

Generate concentration tables, ratios, and basic visualizations, and—if requested—summarize key findings in the context of nicotinate/nicotinamide and NAD pathway biology.

Sample Requirements for Nicotinate and Nicotinamide LC–MS/MS Analysis

For other matrices such as culture media, formulated products, or specialized biological fluids, tailored requirements can be defined during project setup.

What You Receive: Nicotinate and Nicotinamide Analysis Deliverables

Method & run summary: PDF with sample list and groups, run order, and a short method description (prep outline, LC gradient, MS mode, key MRM transitions).

Raw LC–MS/MS files: Vendor-format raw data for standards, QCs, and all study samples.

Processed result tables: Editable spreadsheets (e.g., .xlsx) with peak areas and calculated concentrations for each analyte, linked to sample IDs.

Calibration & QC report: Brief report summarizing calibration curves, QC performance, and any excluded injections or notes.

Optional figures: Simple charts (bar/box plots, group comparisons) for key nicotinate/nicotinamide levels and ratios, if requested.

Representative LC–MS/MS MRM chromatograms of nicotinate, nicotinamide, NAD⁺, and NMN in blank, QC, and experimental samples, showing specific and well-resolved peaks.

NAD⁺ LC–MS/MS calibration curve with seven standards, linear regression and R², and LLOQ/ULOQ markers defining the quantitative range.

Research Applications of Nicotinate and Nicotinamide Profiling

Redox and energy metabolism research

Track NAD precursor pools under metabolic stress or nutrient perturbation.

Drug mechanism and target engagement

Evaluate how small molecules influence nicotinate/nicotinamide utilization and salvage pathways.

Mitochondrial and oxidative stress studies

Relate precursor availability to changes in mitochondrial function and reactive oxygen species handling.

Nutrition and functional ingredient evaluation

Characterize how diets, supplements, or fortification strategies affect vitamin B3–related metabolites.

One-carbon and cofactor network mapping

Integrate nicotinate/nicotinamide data with folate, SAM/SAH, and other cofactor pathways.

Microbiome–host interaction research

Explore how microbial metabolism shapes host pyridine metabolites such as trigonelline or nicotinurate.