Pantothenate & CoA Metabolism Analysis

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Why Analyze the Pantothenate & CoA Metabolic Pathway and Biosynthesis?

Pantothenate (vitamin B5) is the precursor of coenzyme A (CoA), a central cofactor that carries acyl groups in hundreds of biochemical reactions. The pantothenate & CoA metabolic pathway converts pantothenate through a series of phosphorylated intermediates into CoA, linking carbohydrate, lipid and amino acid metabolism.

When this pantothenate & CoA biosynthesis pathway is perturbed, cells rapidly adjust energy production, fatty acid synthesis, and even epigenetic regulation through changes in acetyl-CoA and other acyl-CoA species. Subtle shifts in pathway intermediates can therefore signal:

metabolic stress or nutrient imbalance
drug target engagement in microbes and parasites
functional changes in immune cells and tumors

Targeted quantitative analysis of the pantothenate & CoA metabolic pathway provides pathway-level information that goes far beyond single-metabolite readouts. Our service is designed to capture these changes with high sensitivity and reproducibility, so you can connect experimental interventions to clear biochemical mechanisms.

Pantothenate & CoA Biosynthesis Targeted Metabolomics Solutions

Standard pantothenate & CoA biosynthesis targeted assay: Ready-to-use LC–MS/MS method covering the core pantothenate & CoA biosynthesis pathway.

Custom panel design and expansion: Tailored panel setup or extension, including combination with other targeted metabolomics panels (e.g. central carbon metabolism, amino acids, organic acids, lipids).

Pathway visualization and biological interpretation: Mapping quantitative changes onto the pantothenate & CoA metabolic pathway, with expert comments on affected steps and potential mechanisms.

Integration into broader metabolomics and multi-omics projects: Incorporation of pantothenate & CoA biosynthesis data into larger metabolomics studies or multi-omics workflows.

Pantothenate & CoA Biosynthesis Metabolite Panel and Pathway Coverage

Our pantothenate & CoA biosynthesis targeted metabolomics panel focuses on key intermediates along the vitamin B5–to–CoA route. Representative analytes include, but are not limited to, the following metabolites:

Pantothenate & CoA Biosynthesis Pathway Metabolites

Metabolite	Role in Pantothenate & CoA Biosynthesis
Pantothenate (Vitamin B5)	Precursor for CoA biosynthesis, essential for energy metabolism and acyl transfer reactions.
Pantoic acid	Intermediate in pantothenate biosynthesis. Involved in the conversion of pantoate to pantothenate.
β-Alanine	Combines with pantoate to form pantothenate, crucial in amino acid and energy metabolism.
4'-Phosphopantothenate	First phosphorylated intermediate in the biosynthesis of CoA. Essential for further CoA production.
4'-Phosphopantothenoyl-L-cysteine	Cysteine conjugate intermediate, connecting pantothenate to CoA in the biosynthetic pathway.
Phosphopantetheine	Central intermediate linking pantothenate to CoA, required for CoA activation.
Dephospho-CoA	Key intermediate in CoA biosynthesis, necessary for enzymatic reactions in metabolic pathways.
Coenzyme A (CoA)	Final active cofactor in the biosynthesis of fatty acids, cholesterol, and steroid hormones.

Acyl-CoA Species and Related Metabolites

Metabolite	Role in Pantothenate & CoA Biosynthesis
Acetyl-CoA	The most common acyl-CoA, central to the TCA cycle, fatty acid synthesis, and protein acetylation.
Malonyl-CoA	Precursor for fatty acid synthesis, involved in lipid metabolism and regulation of mitochondrial function.
Palmitoyl-CoA	Intermediate in fatty acid metabolism and mitochondrial function. Used in acylation reactions.
Butyryl-CoA	Involved in fatty acid metabolism, particularly in the catabolism of branched-chain fatty acids.

Upstream Precursors and Related Intermediates

Metabolite	Role in Pantothenate & CoA Biosynthesis
Pantoate	The precursor for pantothenate, essential in the first step of CoA biosynthesis.
Dehydropantoate	Intermediate in the conversion of pantoate to pantothenate, part of the initial biosynthetic steps.

Optional Custom Metabolites

Additional metabolites can be added to the panel based on your research needs:

CoA derivatives (e.g., Acyl-CoA esters) involved in lipid and amino acid metabolism.
Other cofactor-related metabolites such as CoQ10, NAD+, NADH, and FAD, for more comprehensive metabolic pathway analysis.

Why Choose Our Pantothenate & CoA Targeted Metabolomics

Pathway-focused targeted LC–MS/MS
Optimized MRM methods for pantothenate (vitamin B5), CoA and key biosynthetic intermediates, with typical run times of 10–20 minutes per sample for efficient batch analysis.
High sensitivity and wide dynamic range
LLOQ typically in the low-nanomolar range for most pantothenate & CoA biosynthesis intermediates, with 3–4 orders of magnitude linear dynamic range (R² ≥ 0.99).
Excellent precision and reproducibility
Intra-batch QC CVs typically < 10%, inter-batch CVs typically < 15%, supporting robust comparisons across groups and time points.
Quantitative accuracy with isotope-labeled standards
Optional isotope-labeled internal standards and matrix-matched calibration curves, with typical analyte recoveries in the 80–120% range to enhance absolute quantification accuracy.
Low missing-value rates in targeted data
Targeted acquisition and strict QC help keep missing-value rates below 5–10% in well-prepared sample sets, reducing the need for imputation.

Technical Parameters for Pantothenate & CoA Biosynthesis LC–MS/MS Analysis

We utilize a robust, validated LC–MS/MS setup tailored for quantitative analysis of pantothenate, CoA, and related biosynthetic intermediates.

Instrumentation Overview

Component	Model	Function
HPLC System	Agilent 1260 Infinity II	High-performance liquid chromatography for efficient separation of polar metabolites. Stable gradient flow and reproducible retention times support high-throughput analysis.
Mass Spectrometer	Agilent 6495C Triple Quadrupole	Ultra-sensitive LC–MS/MS system optimized for MRM-based targeted metabolomics, capable of detecting analytes at low-nanomolar concentrations with wide dynamic range.

Technical Specifications

Parameter	Typical Performance
LOD (Limit of Detection)	Low nanomolar (nM) range for most analytes
LOQ (Limit of Quantification)	Typically 1–5 nM, depending on matrix and analyte
Linear Dynamic Range	3–4 orders of magnitude (e.g. 0.5–5,000 nM), R² ≥ 0.99
Run Time per Sample	10–20 minutes
Intra-batch Precision	CV < 10% (based on QC replicates)
Inter-batch Precision	CV < 15% (in controlled multi-batch runs)
Quantification Method	External or matrix-matched calibration with internal standards
Calibration Levels	Typically 6–8 levels per compound
Recovery Accuracy	80–120% (when using isotope-labeled standards)

Agilent 6495C Triple Quadrupole (Figure from Agilent)

Agilent 1260 Infinity II HPLC (Fig from Agilent)

Workflow for Pantothenate & CoA Biosynthesis Analysis

Project Consultation

Clarify study goals, models, matrices and target questions for pantothenate & CoA biosynthesis.

Panel Design & Method Setup

Select a standard pantothenate & CoA biosynthesis panel or define a custom panel, then configure LC–MS/MS MRM methods and calibration strategy.

Sample Preparation & QC Setup

Extract polar metabolites with validated protocols and prepare internal standards, calibration standards, blanks and pooled QC samples.

LC–MS/MS Acquisition

Run targeted MRM analysis on triple quadrupole LC–MS/MS systems, injecting QC samples at regular intervals to monitor batch and instrument performance.

Data Processing & Quantification

Integrate peaks, apply calibration and internal standard normalization, and generate curated concentration tables with key QC metrics for pantothenate & CoA pathway metabolites.

Pantothenate & CoA biosynthesis analysis 5-step workflow diagram.

Sample Requirements for Pantothenate & CoA Biosynthesis Projects

Sample type	Recommended amount per sample	Container & labeling	Storage & stability	Special notes
Plasma / Serum	≥ 100–200 µL	Pre-labelled low-bind microcentrifuge tubes	Snap-freeze in liquid N₂; store at −80 °C; avoid >2 freeze–thaw cycles	Collect using EDTA or heparin tubes; centrifuge promptly; separate plasma/serum within 1–2 h.
Whole Blood (by agreement)	≥ 200–300 µL	Pre-labelled cryovials	Snap-freeze and store at −80 °C	Requires prior consultation; anticoagulant type and protocol must be aligned with project goals.
Tissues (animal / plant)	≥ 20–30 mg wet weight	Pre-cooled cryovials or foil-wrapped tissue	Snap-freeze immediately after collection; store at −80 °C	Minimize ischemia time; avoid buffer immersion; record tissue type, location and treatment.
Cultured Cells	≥ 1–5 × 10⁶ cells (pellet, washed)	Pre-labelled low-bind tubes	Snap-freeze cell pellets; store at −80 °C	Wash quickly with ice-cold PBS or saline; remove supernatant completely before freezing.
Biofluids (urine, CSF, others)	≥ 200–500 µL (depending on matrix)	Pre-labelled microcentrifuge tubes	Aliquot, snap-freeze, store at −80 °C	Clarify whether samples are spot, timed, or pooled collections; note any preservatives used.
Microbial Cultures / Pellets	Equivalent to ≥ 20 mg wet biomass	Pre-labelled tubes or cryovials	Snap-freeze and store at −80 °C	Indicate medium, growth phase, OD and treatment; pellet and remove medium quickly on ice.
Extracts (by prior agreement)	≥ 50–100 µL of pre-extracted metabolite solution	LC–MS compatible tubes (e.g., PP or glass)	Store at −80 °C, protected from light if applicable	Provide full extraction solvent composition and protocol; avoid detergents and high salt.
General Shipping Instructions	—	—	Ship on dry ice (minimum 5 kg) to maintain −80 °C conditions	Include a detailed sample list (ID, matrix, treatment, time point); avoid shipment on weekends/holidays.

Recommended Replicates

Cell and microbial experiments: ≥ 6 biological replicates per group
Animal studies: ≥ 8–10 biological replicates per group
Human cohorts: as large as feasible; please contact us to discuss power and design

What You Receive from Our Pantothenate & CoA Biosynthesis Analysis Service

Processed LC–MS/MS data tables
Peak lists and quantitative result tables for pantothenate, CoA and covered intermediates (per sample, per group).
QC performance summary
Brief report with calibration results, linear ranges, basic accuracy and precision (CV) metrics.
Method and settings overview
Concise description of sample preparation, LC conditions, MRM settings and data-processing workflow.
Raw data files (on request)
Vendor-format raw LC–MS/MS files and/or export formats agreed at project setup (research use only).

LC-MS/MS MRM chromatograms of pantothenate and CoA pathway intermediates.

LC–MS/MS MRM Chromatograms for Pantothenate & CoA Intermediates

Calibration curves showing linearity for CoA and major intermediates.

Calibration Curves and Linearity for CoA and Key Intermediates

Applications of Pantothenate & CoA Biosynthesis Targeted Metabolomics

Metabolic Disease and Nutrition

Obesity, insulin resistance, diabetes, NAFLD and other cardiometabolic disorders
Evaluation of vitamin B5 / CoA status and CoA-dependent energy and lipid metabolism
Dietary, nutraceutical or lifestyle intervention studies with metabolic endpoints

Antimicrobial and Antiparasitic Drug Discovery

Target validation and mechanism-of-action studies for pantothenate & CoA pathway enzymes
Screening and optimization of small-molecule pathway inhibitors
PK/PD and resistance studies using pathway intermediates as pharmacodynamic biomarkers

Immune Cell and Cancer Metabolism

Metabolic profiling of T cells, NK cells and myeloid cells during activation or exhaustion
Characterizing tumor reliance on CoA-dependent pathways and acyl-CoA pools
Evaluating metabolic, immuno-oncology or epigenetic therapies affecting cofactor metabolism

Plant, Animal and Agricultural Research

Stress response studies (heat, drought, salinity, multi-stress environments)
Feed and diet optimization in livestock, companion animals and aquaculture
Crop and plant metabolomics where vitamin/cofactor metabolism is a regulatory node

Why measure pantothenate and CoA instead of only downstream metabolites?

Pantothenate→CoA forms a central control point for carbon entry into the TCA cycle, fatty acid synthesis and protein acetylation, so profiling pantothenate, CoA and early intermediates helps distinguish substrate limitation from true pathway block or rerouting—something that is not always clear from lactate, free fatty acids or TCA intermediates alone.

What types of projects benefit most from a pantothenate & CoA panel?

This panel is ideal when you need to understand metabolic control rather than just end products, for example when evaluating pantothenate kinase or CoA-synthesis inhibitors, studying nutrient or hypoxia-induced energy stress, characterizing immune cell activation or tumor metabolic rewiring, or probing microbial and parasite viability that depends on pantothenate/CoA biosynthesis.

Which sample types are suitable for CoA and acyl-CoA analysis?

Metabolically active tissues (e.g., liver, muscle, heart, tumors), activated immune cells, microbial or parasite pellets, and well-processed plasma/serum or cell-free supernatants are commonly used; the critical factor is rapid quenching and extraction with cold organic solvents to stabilize labile CoA and acyl-CoA thioesters and minimize post-sampling turnover.

Can this panel be applied to non-mammalian systems such as microbes or plants?

Yes, pantothenate and CoA biosynthesis is highly conserved in bacteria, fungi, parasites and plants, so with matrix-specific optimization and validation, the same LC–MS/MS principles can be used to interrogate CoA-pathway flux in microbial cultures, plant tissues and host–pathogen models.

How is analytical specificity for individual CoA and acyl-CoA species ensured?

Specificity is achieved by combining appropriate chromatographic separation with carefully optimized MRM transitions for the CoA backbone and acyl fragments, retention-time windows, and where available, stable isotope internal standards, allowing CoA, its biosynthetic intermediates and structurally similar acyl-CoAs to be resolved in complex matrices.

The Brain Metabolome Is Modified by Obesity in a Sex-Dependent Manner

Norman, J. E., Milenkovic, D., Nuthikattu, S., & Villablanca, A. C.

Journal: International Journal of Molecular Sciences

Year: 2024

Quantifying forms and functions of intestinal bile acid pools in mice

Sudo, K., Delmas-Eliason, A., Soucy, S., Barrack, K. E., Liu, J., Balasubramanian, A., ... & Sundrud, M. S.

Journal: bioRxiv

Year: 2024

Thermotolerance capabilities, blood metabolomics, and mammary gland hemodynamics and transcriptomic profiles of slick-haired Holstein cattle during mid lactation in Puerto Rico

Contreras-Correa, Z. E., Sánchez-Rodríguez, H. L., Arick II, M. A., Muñiz-Colón, G., & Lemley, C. O.

Journal: Journal of Dairy Science

Year: 2024