Alpha-Ketoglutaric Acid Analysis Service

Alpha-Ketoglutaric Acid (α-KG), a vital TCA cycle metabolite, is crucial for cancer, mitochondrial, and aging research. Creative Proteomics offers precise α-KG detection using LC-MS/MS, UHPLC, and GC-MS, with sensitivity down to 0.01 ng/mL. Services include quantification, metabolic profiling, and isotope tracing, supporting IDH mutation studies, microbial optimization, and dietary research. Integrated multi-omics and regulatory-compliant reporting deliver reliable data for research and industry.

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What is α-Ketoglutaric Acid?

α-Ketoglutaric acid (α-KG) is a pivotal intermediate in the tricarboxylic acid (TCA) cycle, playing essential roles in cellular energy production, amino acid synthesis, and epigenetic regulation. As a key metabolic hub, it serves as a substrate for multiple enzymes (e.g., α-KG-dependent dioxygenases) and participates in redox balance maintenance. Dysregulation of α-KG levels is linked to mitochondrial disorders, cancer progression, and aging-related pathologies. Accurate quantification of α-KG is critical for understanding metabolic flux dynamics in research areas ranging from oncology to nutraceutical development. Creative Proteomics can quantify and characterize α-ketoglutaric acid in various biological samples

α-Ketoglutaric Acid Analysis Services Provided by Creative Proteomics

Quantitative Measurement: Accurate determination of α-Ketoglutaric acid concentrations in biological samples such as serum, plasma, urine, and tissues.

Metabolomics Profiling: Comprehensive analysis of α-Ketoglutaric acid alongside other metabolites to assess metabolic pathways and alterations.

Pathway Analysis: Detailed insights into the role of α-Ketoglutaric acid in metabolic and disease pathways, including cancer and neurodegeneration.

Isoform Detection: Identification and quantification of α-Ketoglutaric acid isoforms and related derivatives.

Isotopic Labeling: Tracking the fate of α-Ketoglutaric acid in metabolic studies using stable isotopes.

Custom Solutions: Tailored analysis services designed to meet specific research or clinical needs.

List of Detected α-Ketoglutaric Acid and Related Compounds

Compound	Detection Method	Application Area
α-Ketoglutaric Acid	LC-MS, GC-MS, NMR	Metabolomics, Biochemistry
Glutamate	LC-MS, HPLC	Neurochemistry, Neurodegeneration
Glutamine	LC-MS, HPLC	Metabolism, Clinical Research
Succinic Acid	GC-MS, LC-MS	Energy Metabolism, Cancer Studies
Citrate	HPLC, LC-MS	TCA Cycle, Metabolic Studies
Malate	LC-MS, GC-MS	Biochemistry, Metabolism Studies

Methods for α-Ketoglutaric Acid Detection

High-Performance Liquid Chromatography (HPLC)

Instrument: Agilent 1260 Infinity

Key Parameters:

Column: A C18 column (e.g., Waters BEH C18, 1.7 µm) is recommended for baseline separation of α-KG from isobaric metabolites 4.
Detection: UV detection at 210 nm or diode array detectors (DAD) are commonly employed.

Liquid Chromatography-Mass Spectrometry (LC-MS)

Instrument: Thermo Scientific Q Exactive HF-X Orbitrap.

Key Parameters:

Ionization Mode: Electrospray ionization (ESI) in negative mode, targeting the [M-H]⁻ ion of α-KG at m/z 145.01.
Quantification: A dynamic range of 0.1–500 µM is achievable with isotope-labeled internal standards (e.g., ¹³C-α-KG) for calibration.

Gas Chromatography-Mass Spectrometry (GC-MS)

Instrument: Agilent 7890B GC with Agilent 5977A MSD

Key Parameters:

Derivatization: Trimethylsilyl (TMS) derivatives improve detection sensitivity.
Separation: A DB-5MS column (30 m × 0.25 mm) with a temperature gradient (50°C to 300°C) achieves baseline separation of α-KG from organic acids like fumarate or malate.

Agilent 1260 Infinity II HPLC (Fig from Agilent)

Thermo Fisher Scientific Q Exactive™ HF-X Hybrid Quadrupole-Orbitrap™ Mass Spectrometer

Q Exactive™ HF-X Hybrid Quadrupole-Orbitrap™ Mass Spectrometer (Fig from Thermo Fisher)

Agilent 7890B Gas chromatography system (Fig from Agilent)

Advantages of Our α-Ketoglutaric Acid Detection

Ultra-High Sensitivity: LC-MS/MS (SCIEX Triple Quad 6500+) achieves detection limits as low as 0.01 ng/mL, ideal for trace-level α- ketoglutaric acid quantification in complex matrices like CSF or microbial lysates.
Multi-Platform Validation: Cross-verified results using UHPLC (Waters ACQUITY) and GC-MS (Agilent 8890/5977B) ensure method robustness and data reproducibility.
Matrix Flexibility: Pre-validated protocols for diverse samples—serum, tissue, cell cultures, urine, and soil extracts—with recovery rates >95%.
Isotope-Resolved Precision: ¹³C/²H-labeled α- ketoglutaric acid tracing via GC-MS enables accurate metabolic flux analysis in dynamic systems (e.g., microbial fermentation).

Sample Requirements for α-Ketoglutaric Acid Analysis

Sample Type	Minimum Sample Volume	Recommended Sample Storage	Sample Preparation Requirements
Serum	100 µL	-80°C (long-term storage)	Centrifuge to separate plasma from cells.
Plasma	100 µL	-80°C	Centrifuge and separate from cells.
Urine	1 mL	-80°C	No special preparation needed, avoid contamination.
Tissue (Frozen)	50 mg	-80°C	Homogenize tissue for uniform analysis.
Cultured Cells	1-5 million cells	-80°C	Wash cells in cold PBS before freezing.
Saliva	1 mL	-80°C	Collect in sterile containers.

Applications of α-Ketoglutaric Acid Assay

Metabolic Disorder Diagnosis

Detect disruptions in the TCA cycle, aiding in the diagnosis of mitochondrial diseases.

Cancer Metabolism

Study metabolic reprogramming and the Warburg effect in cancer cells.

Neurodegenerative Diseases

Investigate metabolic dysfunctions in Alzheimer's and Parkinson's.

Nutritional Assessments

Evaluate dietary impacts on amino acid metabolism and nitrogen balance.

Aging Research

Analyze α-Ketoglutaric acid's role in cellular senescence and mitochondrial function.

Disease Models

Track metabolic changes in cellular models for various diseases.

Demo Result of Targeted Metabolomics Service

Figures come from (Li, Y.et.al, Sci Rep,2023)

Principal Component Analysis (PCA) chart showing the distribution of samples across principal components

Quickly show the distribution of samples across principal components.

Orthogonal Partial Least Squares Method-Discriminant Analysis (PLS-DA) point cloud diagram illustrating the separation of sample groups in a multidimensional space

OPLS-DA point cloud diagram

Illustrate the separation of sample groups in a multidimensional space.

Plot of multiplicative change volcanoes

Volcano plot depicting multiplicative changes in metabolite levels.

Dendrogram representing hierarchical clustering of samples.

Sample Hierarchical Clustering

Each column in the figure represents a sample

FAQ of α-Ketoglutaric Acid Analysis

Can you distinguish α-KG from its isomers or derivatives?

Yes. Our LC-MS/MS method achieves baseline separation of α-KG from structurally similar metabolites (e.g., D/L-2-hydroxyglutarate) using high-resolution mass spectrometry (HRMS, 140,000 FWHM)

What bioinformatics support is provided?

Reports include:

α-KG/pyruvate ratios and TCA cycle flux maps.
Pathway enrichment analysis via MetaBridge™, integrating transcriptomic/proteomic datasets

What sample types are compatible with α-KG analysis?

We support a wide range of biological matrices, including serum/plasma, cell lysates, tissue homogenates, urine, and microbial cultures. For specialized matrices like cerebrospinal fluid (CSF) or soil extracts, customized protocols are available to ensure >95% recovery rates

Learn about other Q&A.

α-Ketoglutaric Acid Analysis Case Study

Article

Improved Method for Quantifying Sugars and Organic Acids in Tomato Fruits

Publications

Here are some publications in proteomics research from our clients:

The Suppression of the KRAS G12D-Nrf2 Axis Shifts Arginine into the Phosphocreatine Energy System in Pancreatic Cancer Cells. 2023. http://dx.doi.org/10.2139/ssrn.4318051
Resting natural killer cell homeostasis relies on tryptophan/NAD+ metabolism and HIF‐1α. 2023. https://doi.org/10.15252/embr.202256156
Mechanisms underlying neonate-specific metabolic effects of volatile anesthetics. 2021. https://doi.org/10.7554/eLife.65400
Choleoeimeria pogonae Alters the Bile Acid Composition of the Central Bearded Dragon (Pogona vitticeps). 2021. https://doi.org/10.5818/JHMS-D-20-00009
Anti-inflammatory activity of black soldier fly oil associated with modulation of tlr signaling: A metabolomic approach. 2023. https://doi.org/10.3390/ijms241310634

More Publications

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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