Isovaleric Acid Analysis Service

Creative Proteomics provides isovaleric acid analysis using advanced LC-MS/MS and GC-MS platforms. We offer comprehensive metabolite profiling, enabling researchers and industries to study metabolic pathways, monitor fermentation processes, ensure product quality, and assess microbial activity. Our solutions deliver actionable insights for pharmaceutical, food and beverage, biotechnology, and environmental applications.

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What We Provide
Advantages
Technology Platform
Sample Requirements
FAQ
Publications

What is Isovaleric Acid?

Isovaleric acid (3-methylbutanoic acid) is a branched-chain short-chain fatty acid (BCFA) produced during leucine catabolism. This metabolite is integral to energy metabolism and microbial fermentation processes. Beyond its biochemical significance, isovaleric acid is a key flavor compound in fermented foods (e.g., cheese, wine) and a critical indicator in microbiome studies, where it reflects microbial activity and substrate utilization. Its quantification is essential for optimizing industrial processes and advancing research in metabolic pathways and microbial ecology.

Structure of isovaleric acid

Isovaleric Acid Analysis Service Offered by Creative Proteomics

Isovaleric Acid Quantification: Accurate measurement of isovaleric acid in biological samples (serum, plasma, tissues) and industrial samples (fermented foods, beverages).
Metabolic Pathway Mapping: Analysis of leucine degradation intermediates, including β-hydroxyisovaleric acid and isovaleryl-CoA derivatives, to study metabolic pathways.
Microbiome Profiling: Quantification of microbial fermentation products to study gut, soil, or industrial microbiome dynamics.
Food & Beverage Quality Control: Monitoring of isovaleric acid and related compounds to ensure product consistency and optimize fermentation protocols.
Pharmaceutical Research: Tracking metabolic shifts in drug development and toxicology studies.
Environmental Monitoring: Analysis of isovaleric acid levels as a biomarker for organic compound degradation in ecosystems.

List of Isovaleric Acid and Related Metabolites

List of Isovaleric Acid and Related Metabolites
Isovaleric Acid (IVA)	Isovaleryl-CoA	3-Methylcrotonyl-CoA	3-Hydroxyisovaleric Acid	2-Methylbutyryl-CoA
Methylmalonic Acid	Acetyl-CoA	Isovalerylglycine	3-Methylbutyrate	2-Methylbutyrate
2-Hydroxyisovaleric Acid	2-Ketoisovaleric Acid	2-Methylbutanoyl-CoA	Valeric Acid

Advantages of Isovaleric Acid Assay

High Sensitivity and Precision: Detection limit as low as 1 ng/mL with coefficients of variation (CV) below 5% using LC-MS/MS or GC-MS.
Comprehensive Metabolite Coverage: Simultaneous quantification of isovaleric acid and over 20 related metabolites with isotope-labeled standards.
Advanced Analytical Platforms: High-resolution analysis using Triple Quadrupole and Orbitrap mass analyzers with ppm-level accuracy.
Customized Solutions: Tailored assay designs and expert support for data interpretation and metabolic pathway analysis.
High Throughput Capabilities: Capable of analyzing up to 500 samples per batch for large-scale studies.
Low Matrix Interference: Advanced sample preparation techniques minimize matrix effects, ensuring accurate results.
Stable and Reproducible Results: Strict quality control procedures ensure inter-batch variability below 10%.
Method Validation Support: Full method development and validation services available, including linearity, accuracy, precision, and recovery assessments.
Data Visualization and Interpretation: Comprehensive data analysis with graphical representations for clear interpretation of metabolic changes.

Technology Platforms for Isovaleric Acid Analysis Service

HPLC-UV (High-Performance Liquid Chromatography with UV Detection)

Agilent 1260 Infinity II HPLC (Figure from Agilent)

GC-MS (Gas Chromatography-Mass Spectrometry)

7890B Gas Chromatograph + 5977 Single Quadrupole

Agilent 7890B-5977B (Figure from Agilent)

LC-MS/MS (Liquid Chromatography-Tandem Mass Spectrometry)

SCIEX Triple Quad™ 6500+ (Figure from Sciex)

LC-MS

Q Exactive™ Plus Hybrid Quadrupole-Orbitrap™ Mass Spectrometer (Figure from Thermo)

Sample Requirements for Isovaleric Acid Analysis Service

Sample Type	Minimum Volume	Storage Condition	Additional Notes
Plasma	50 µL	-80°C	Use EDTA or heparin as anticoagulant
Serum	50 µL	-80°C	Avoid hemolysis for accurate results
Urine	100 µL	-80°C	First morning urine preferred
Tissue Homogenate	50 mg or 100 µL	-80°C	Flash freeze in liquid nitrogen
Fermentation Broth	1 mL	-20°C to -80°C	Ensure proper mixing before sampling
Cell Culture Medium	1 mL	-80°C	Collect in sterile tubes
Feces	50 mg	-80°C	Store without preservatives
Soil or Environmental Sample	1 g	-20°C	Air-dry or freeze immediately
Microbial Culture	1 mL	-80°C	Centrifuge to separate supernatant if needed

Applications of Isovaleric Acid Assay Service

Metabolic Research

Investigate leucine catabolism, related metabolic pathways, and biochemical regulation for a deeper understanding of metabolism.

Microbiome and Environmental Studies

Analyze microbial fermentation products in gut microbiomes, soil, and water samples to evaluate microbial activity and ecosystem dynamics.

Food and Beverage Quality Control

Monitor flavor compounds and volatile fatty acids to ensure product consistency, detect spoilage, and optimize fermentation processes.

Pharmaceutical and Biomedical Research

Track metabolic changes in drug development, toxicology studies, and biomarker discovery.

Industrial Biotechnology

Assess fermentation performance, monitor byproducts, and optimize bioprocessing efficiency for industrial-scale production.

Agricultural and Nutritional Studies

Evaluate the effects of dietary interventions, livestock health, and agricultural practices on metabolic outputs and microbial activity.

Demo

GC-MS chromatogram showing SCFA separation, quantification of SCFA levels, and corresponding mass spectra with molecular ions for Ac, PA, Iso-BA, BA, and Iso-VA.

(A) GC-MS total ion chromatogram for the separation of SCFAs, including acetic acid (Ac), propionic acid (PA), 2 H7-butyric acid (*) as an internal standard, isobutyric acid (Iso-BA), butyric acid (BA), and isovaleric acid (Iso-VA). (B) Quantification of SCFA levels (μg/mL). (C) Mass spectra showing molecular ions (m/z) for Ac, PA, Iso-BA, BA, and Iso-VA (Kao et al., 2017).

FAQ of Isovaleric Acid Analysis Service

How do you handle complex sample matrices like fermented foods or microbial lysates to ensure accurate quantification of isovaleric acid?

We employ matrix-specific pretreatment protocols to isolate isovaleric acid from interfering substances. For high-fat matrices (e.g., cheese), we use solid-phase extraction (SPE) with C18 cartridges and lipid-absorbent resins to remove triglycerides. For microbial lysates, enzymatic digestion (e.g., lysozyme treatment) followed by centrifugation ensures complete cell lysis and metabolite release. All workflows are validated with isotope-labeled internal standards (d₅-isovaleric acid) to correct for recovery efficiency and matrix effects.

What experimental design recommendations do you have for longitudinal studies tracking isovaleric acid dynamics in microbiome research?

For longitudinal studies, we recommend:

Consistent sampling intervals: Collect samples at fixed timepoints (e.g., 0, 24, 48 hours post-intervention) to capture microbial metabolic shifts.
Parallel negative controls: Include sterile culture media or placebo-treated samples to distinguish background noise.
Triplicate aliquots: Minimize batch variability by splitting samples into triplicates for independent analysis.
Metadata integration: Provide detailed logs of environmental conditions (pH, temperature) and microbial load to correlate with isovaleric acid levels.

Can you quantify trace-level isovaleric acid in air or headspace samples from industrial fermentation reactors?

Yes. We utilize thermal desorption tubes (Tenax TA/Carbograph adsorbents) to capture volatile isovaleric acid from gas-phase samples. Post-collection, analytes are desorbed at 300°C and analyzed via Agilent 7890B GC-MS with a DB-FFAP column, achieving a detection limit of 0.5 ppb. Calibration curves are prepared using NIST-traceable standards to ensure industrial-grade precision.

How do you differentiate isovaleric acid from structurally similar branched-chain fatty acids (e.g., isobutyric acid) in mixed samples?

Our high-resolution LC-MS/MS (Sciex 6500+) method separates isomers using a HILIC column (ZIC-cHILIC, 2.1 × 150 mm) with a 15-minute gradient (0.1% formic acid in acetonitrile/water). Structural confirmation is achieved via MS/MS fragmentation patterns (e.g., m/z 102 → 57 for isovaleric acid vs. m/z 88 → 43 for isobutyric acid). This dual-layer specificity eliminates cross-reactivity.

Learn about other Q&A.

Isovaleric Acid Analysis Service Case Study

Article

Quantitative Determination of Short-Chain Fatty Acids in Human Serum by LC-MS

Publications

Here are some of the metabolomics-related papers published by our clients:

A non-probiotic fermented soy product reduces total and ldl cholesterol: A randomized controlled crossover trial. 2021. https://doi.org/10.3390/nu13020535
Resting natural killer cell homeostasis relies on tryptophan/NAD+ metabolism and HIF‐1α. 2023. https://doi.org/10.15252/embr.202256156
Enhance trial: effects of NAD3® on hallmarks of aging and clinical endpoints of health in middle aged adults: a subset analysis focused on blood cell NAD+ concentrations and lipid metabolism. 2022. https://doi.org/10.3390/physiologia2010002
Dimethyl fumarate treatment restrains the antioxidative capacity of T cells to control autoimmunity. 2021. https://doi.org/10.1093/brain/awab307
Function and regulation of a steroidogenic CYP450 enzyme in the mitochondrion of Toxoplasma gondii. 2023. https://doi.org/10.1371/journal.ppat.1011566

More Publications

References

Kao, Ming‐Shan, et al. "Microbiome precision editing: Using PEG as a selective fermentation initiator against methicillin‐resistant Staphylococcus aureus." Biotechnology journal 12.4 (2017). https://doi.org/10.1002/biot.201600399
Minkler, Paul E., et al. "Selective and accurate C5 acylcarnitine quantitation by UHPLC–MS/MS: Distinguishing true isovaleric acidemia from pivalate derived interference." Journal of Chromatography B 1061 (2017): 128-133. https://doi.org/10.1016/j.jchromb.2017.07.018

Metabolomics Sample Submission Guidelines

Download our Metabolomics Sample Preparation Guide for essential instructions on proper sample collection, storage, and transport for optimal experimental results. The guide covers various sample types, including tissues, serum, urine, and cells, along with quantity requirements for untargeted and targeted metabolomics.

Download

Metabolomics Sample Submission Guidelines

* For Research Use Only. Not for use in diagnostic procedures.

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From Our Clients

"I recently used their proteomics service for a project analyzing protein interactions in yeast models. The team was very responsive and helped clarify the methodology they employed, which made me feel confident in the results. The data quality was solid, with clear identification of several key proteins involved in our study. Their thorough analysis enabled me to pinpoint specific interactions that I hadn't considered before, which significantly improved the direction of my research. I appreciate their professionalism and support throughout the process."

Sarah Thompson, University of California, Berkeley

"Our lab collaborated with them on a project studying cancer biomarkers. The proteomics analysis provided was detailed and focused, specifically highlighting the differential expression of proteins between healthy and tumor samples. Their clear explanations of the data helped my team understand the biological implications. I also appreciated their willingness to revise the reports based on our feedback, ensuring that we had everything we needed for our publication. This collaborative spirit was invaluable."

Emily Rodriguez, Stanford University

"Our lab worked with them on a project studying the effects of diet on gut microbiota using proteomics. They used a label-free quantification method to analyze proteins in fecal samples before and after dietary intervention. The results showed significant changes in protein expression linked to microbial activity. This was pivotal for our hypothesis about diet-microbiota interactions. The clarity of their data presentation made it easy for our team to integrate these findings into our ongoing research."

Dr. Lisa Wong, University of Toronto

"My experience with Creative Proteomics during the mass spectrometry analysis was excellent. We sent in human saliva and mouse brain tissue samples, which they expertly analyzed using both LC-MS and GC-MS techniques. The results were invaluable, revealing key metabolites in the saliva and identifying biomarkers linked to brain function in the brain tissue."

Dr. Emily Carter, Senior Research Scientist

"The overall service from Creative Proteomics was outstanding. They made the entire process seamless and efficient, allowing us to focus on our research. We worked with leaf and root samples from various Arabidopsis genotypes for targeted metabolomics analysis. Their thorough profiling of primary and secondary metabolites gave us important insights into how the plants respond metabolically to environmental stress."

Dr. Laura Henderson, Plant Physiologist

"We had a pleasant collaboration with Creative Proteomics on mass spectrometry analysis of lipids. They conducted a detailed analysis of lipid species, providing us with important insights into lipid metabolism and its relationship with metabolic syndrome disease states."

Dr. Sarah Mitchell, Research Scientist

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