Acylcarnitine Analysis Service

Creative Proteomics offers high-throughput, ultra-sensitive acylcarnitine analysis via LC-MS/MS to support lipid metabolism research, mitochondrial function studies, and metabolic flux profiling. Our service delivers absolute quantification of 30+ acylcarnitines across diverse sample types—plasma, tissue, cells—helping researchers uncover metabolic alterations with precision and speed.

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  • Service We Provide
  • List of Acylcarnitine
  • Advantages
  • Workflow
  • Methods
  • Results and Data Analysis
  • Sample Requirements
  • FAQ

What are Acylcarnitine?

Acylcarnitines are a class of esters formed from carnitine and fatty acids, playing a pivotal role in mitochondrial β-oxidation and energy homeostasis. These metabolites act as shuttles, enabling the transport of fatty acyl groups across the inner mitochondrial membrane for subsequent oxidation. Variations in acylcarnitine levels are directly linked to disruptions in lipid metabolism, mitochondrial function, and amino acid catabolism.

Their unique structural diversity—ranging from short-chain (e.g., acetylcarnitine) to long-chain forms (e.g., palmitoylcarnitine)—makes acylcarnitines reliable indicators of metabolic flux across multiple physiological and pathological states.

Why Analyze Acylcarnitines?

Accurate and comprehensive acylcarnitine analysis enables researchers to:

Creative Proteomics empowers you with a full-spectrum acylcarnitine analysis service, generating reproducible and quantitative insights with high analytical depth.

Acylcarnitine Analysis in Creative Proteomics

Targeted Quantification of Acylcarnitines

Quantify over 30 acylcarnitines (C2–C26) including short-, medium-, long-chain, and unsaturated species.

Hydroxy- and Dicarboxylic Acylcarnitine Profiling

Detect hydroxylated and dicarboxylated acylcarnitines for oxidative and mitochondrial metabolism studies.

Branched-Chain Acylcarnitine Profiling

Analyze acylcarnitines from BCAA metabolism, such as isovaleryl- and methylbutyryl-carnitine.

Multi-Matrix Sample Support

Compatible with plasma, serum, urine, tissues, and cultured cells for flexible experimental designs.

Statistical & Differential Analysis

Includes fold change, CV%, PCA, HCA, and PLS-DA for comprehensive data interpretation.

Metabolic Pathway Integration

Map acylcarnitine data to β-oxidation, carnitine shuttle, and amino acid degradation pathways.

List of Detected Acylcarnitine

ClassRepresentative CompoundsTypical Carbon ChainsRelated Metabolic Pathways
Free CarnitineCarnitine (C0)C0Carnitine shuttle, fatty acid transport
Short-Chain AcylcarnitinesAcetylcarnitine (C2), Propionylcarnitine (C3), Butyrylcarnitine (C4), Isobutyrylcarnitine (C4-i), Valerylcarnitine (C5), Isovalerylcarnitine (C5-i), Tiglylcarnitine (C5:1)C2–C5Fatty acid β-oxidation, branched-chain amino acid catabolism
Medium-Chain AcylcarnitinesHexanoylcarnitine (C6), Octanoylcarnitine (C8), Decanoylcarnitine (C10), Dodecanoylcarnitine (C12)C6–C12Mitochondrial FAO, peroxisomal oxidation
Long-Chain AcylcarnitinesMyristoylcarnitine (C14), Palmitoylcarnitine (C16), Stearoylcarnitine (C18), Oleoylcarnitine (C18:1), Linoleoylcarnitine (C18:2), Arachidoylcarnitine (C20)C14–C20+Carnitine–acylcarnitine translocase activity
Hydroxy-Acylcarnitines3-Hydroxybutyrylcarnitine (C4-OH), 3-Hydroxyhexanoylcarnitine (C6-OH), 3-Hydroxyoctanoylcarnitine (C8-OH), 3-Hydroxypalmitoylcarnitine (C16-OH)C4–C16-OHKetone body metabolism, oxidative stress marker
Unsaturated AcylcarnitinesHexenoylcarnitine (C6:1), Octenoylcarnitine (C8:1), Decenoylcarnitine (C10:1), Palmitoleylcarnitine (C16:1), Oleylcarnitine (C18:1), Linoleylcarnitine (C18:2)C6:1–C18:2FA desaturation and elongation
Dicarboxylic AcylcarnitinesSuccinylcarnitine (C4-DC), Glutarylcarnitine (C5-DC), Adipoylcarnitine (C6-DC), Malonylcarnitine (C3-DC), MethylmalonylcarnitineC3–C6-DCOrganic acid metabolism, amino acid degradation
Branched-Chain AcylcarnitinesIsobutyrylcarnitine (C4-i), Isovalerylcarnitine (C5-i), 2-Methylbutyrylcarnitine (C5-m), 3-Methylglutarylcarnitine (C6-m)C4–C6-branchedLeucine, isoleucine, valine metabolism
Others / Rare SpeciesLaurylcarnitine (C12), Behenoylcarnitine (C22), Cerotoylcarnitine (C26), Hydroxydodecanoylcarnitine (C12-OH), Nonanoylcarnitine (C9)C9–C26, modifiedExploratory biomarker research

Why Choose Our Acylcarnitine Assay?

  • LOD as low as 0.05 ng/mL: Ultra-sensitive detection using Agilent 6495C ensures reliable quantification of low-abundance metabolites.
  • Quantitative Range >10⁵-fold: Enables simultaneous profiling of trace and high-abundance acylcarnitines without signal saturation.
  • >30 Acylcarnitines Covered: Includes short-, medium-, long-chain, hydroxy-, dicarboxylic, branched, and unsaturated species.
  • <10% CV Across Batches: Stable isotope-labeled internal standards ensure high reproducibility and accuracy.
  • Sample-Efficient Workflow: Requires only 100 µL plasma or 20 mg tissue, ideal for limited or precious samples.
  • High Throughput: Supports 80–100 samples per day, enabling efficient large-scale or multi-condition studies.

How Creative Proteomics Provides Acylcarnitine Assay?

Workflow for Acylcarnitine Analysis

What Methods are Used for Our Acylcarnitine Analysis?

Agilent 6495C Triple Quadrupole LC-MS/MS: Used for targeted, ultra-sensitive quantification of short-, medium-, and long-chain acylcarnitines. Ideal for both high- and low-abundance analytes.

Agilent 1260 Infinity II HPLC: Serves as the front-end chromatographic separation system, enabling excellent peak resolution across diverse acylcarnitine classes.

Agilent 6495C Triple Quadrupole

Agilent 6495C Triple Quadrupole (Figure from Agilent)

Agilent 1260 Infinity II HPLC

Agilent 1260 Infinity II HPLC (Fig from Agilent)

Acylcarnitine Analysis Service: Results and Data Analysis

Quantitative Data Output

Absolute Quantification: Each acylcarnitine species—short-, medium-, and long-chain—is reported with precise concentration values (e.g., nmol/mL or μmol/g), based on calibration with stable isotope-labeled internal standards.

High Sensitivity and Reproducibility: Limits of detection reach femtomolar levels. Coefficient of variation (CV) is maintained below 15%, ensuring consistency across biological replicates.

Comprehensive Output Format: All quantitative results are provided in raw and processed forms, including:

  • LC-MS/MS raw chromatograms
  • MRM transition tables
  • Standard curve data
  • Normalized data matrices
High-throughput acylcarnitine profiling using UHPLC-MS/MS within 10 minutes.

Rapid quantification of hundreds of acylcarnitines across diverse biological matrices using UHPLC-MS/MS in under 10 minutes (Zhang, Jingxian, et al., 2024).

Product ion spectra and proposed fragmentation patterns for valeryl-carnitine (C5:0-CN) (A) and 3-methylglutaryl-carnitine (C5-M-DC-CN) (B).

Product ion spectra and proposed fragmentation patterns for valeryl-carnitine (C5:0-CN) (A) and 3-methylglutaryl-carnitine (C5-M-DC-CN) (B) (Giesbertz, Pieter, et al., 2015).

Four-panel image showing mass spectrum, heatmap, volcano plot, and PCA biplot from acylcarnitine analysis.

Representative Results of Acylcarnitine Analysis. Includes a mass spectrum, heatmap, volcano plot, and PCA biplot showing acylcarnitine distribution and group differences.

Statistical and Multivariate Analysis

Descriptive Statistics: Summary tables include mean, standard deviation, and CV% for each compound across all samples.

Comparative Analysis: Fold change and significance testing (p-value, t-test, ANOVA) are included to support differential expression assessments.

Advanced Pattern Recognition: Optional multivariate methods such as:

  • Principal Component Analysis (PCA)
  • Hierarchical Clustering (HCA)
  • PLS-DA for sample classification
    These analyses provide insight into metabolic patterns and group-specific trends.

Pathway Mapping and Biological Interpretation

Pathway-Level Integration: Detected acylcarnitines are mapped to relevant biological pathways including:

  • Mitochondrial β-oxidation
  • Carnitine shuttle system
  • Branched-chain fatty acid and amino acid metabolism

Metabolic Visualization: Results are presented with annotated pathway diagrams, heatmaps, and volcano plots to support biological interpretation.

Bioinformatics Support (Optional): Advanced analyses such as pathway enrichment, correlation network analysis, and multi-omics integration (with transcriptomics or proteomics data) are available upon request.

Custom Report Delivery: Final reports include all data files (raw and processed), visual figures, and interpretation summaries in a publication-ready format.

Explore our Lipidomics Solutions brochure to learn more about our comprehensive lipidomics analysis platform.

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What Our Acylcarnitine Analysis Used For

Metabolic Research

Quantitative insight into lipid metabolism and mitochondrial activity.

Nutritional Science

Assessing the metabolic effects of dietary fats or supplements.

Sports Physiology

Monitoring energy metabolism during endurance or high-intensity exercise.

Toxicology Studies

Evaluating metabolic disruption due to xenobiotics or stressors.

Aging Research

Investigating age-associated shifts in mitochondrial substrate utilization.

Comparative Biology

Species-specific studies of fatty acid oxidation across models.

Sample Requirements for Acylcarnitine Analysis Solutions

Sample TypeRequired AmountStorage & Transport Notes
Plasma / Serum≥ 100 μLCollect using EDTA or heparin tubes; avoid hemolysis. Store at -80°C and ship on dry ice.
Cell Pellet≥ 1 × 10⁷ cellsWash with PBS, centrifuge, snap-freeze in liquid nitrogen. Store at -80°C.
Animal Tissue≥ 50 mgFreeze immediately after collection. Avoid preservatives. Store at -80°C, ship on dry ice.
Cell Culture Medium≥ 500 μLUse serum-free medium if possible. Pre-clear by centrifugation. Store at -80°C.
Plant Tissue≥ 100 mg (fresh/frozen)Rinse, blot dry, freeze or freeze-dry. Store at -80°C or in vacuum-sealed bags.
Bacterial Biomass≥ 5 × 10⁸ cellsPellet by centrifugation, wash with PBS. Snap-freeze and store at -80°C.
Fungal Mycelia≥ 100 mg (wet weight)Harvest, rinse if needed, freeze immediately. Store at -80°C.

FAQs for Acylcarnitine Analysis Service

What types of biological samples can be analyzed?

We accept a wide range of sample types, including plasma, serum, tissue (e.g., liver, muscle), cell lysates, and cultured media. For tissue samples, a minimum of 20 mg is required, while liquid samples (e.g., plasma) need only 100 µL.

How should samples be prepared and stored prior to shipment?

Samples should be snap-frozen in liquid nitrogen immediately after collection to preserve metabolite integrity. Ship them on dry ice via overnight courier. Avoid repeated freeze-thaw cycles, as this may degrade acylcarnitines.

Can I compare acylcarnitine levels across different sample types (e.g., plasma vs. tissue)?

Yes, our normalization workflows account for matrix-specific differences. Concentrations are reported in standardized units (e.g., nmol/mL for biofluids, μmol/g for tissues), enabling cross-comparison.

How do I interpret elevated levels of specific acylcarnitines (e.g., C16-OH)?

Elevations in hydroxylated (e.g., C16-OH) or dicarboxylic species often indicate mitochondrial oxidative stress or β-oxidation bottlenecks. We provide pathway mapping and biological context in the report, but consultation with a metabolic specialist is recommended for clinical or complex cases.

What if my samples contain very low concentrations of acylcarnitines?

Our LC-MS/MS platform achieves a limit of detection (LOD) as low as 0.05 ng/mL, ensuring reliable quantification even for trace metabolites. If working with extremely low-abundance samples (e.g., cerebrospinal fluid), we can optimize pre-concentration steps upon request.

Can you integrate acylcarnitine data with other omics datasets (e.g., proteomics)?

Yes, we offer optional bioinformatics support for multi-omics integration. This includes correlating acylcarnitine profiles with proteomic pathways (e.g., mitochondrial enzymes) or transcriptomic data to uncover mechanistic insights.

Are there batch effects in large-scale studies?

Our use of stable isotope-labeled internal standards minimizes batch variability (<10% CV). For multi-batch projects, we apply cross-batch normalization algorithms to ensure data consistency.

How are unsaturated acylcarnitines differentiated from saturated ones?

Chromatographic separation combined with MS/MS fragmentation distinguishes isomers (e.g., C18:1 vs. C18:2) based on retention time and unique product ion patterns.

Do you provide guidance on follow-up experiments?

Yes, we offer complimentary consultations to help design validation experiments (e.g., stable isotope tracing for flux analysis) or suggest complementary assays (e.g., free fatty acid profiling) based on your results.

Are results compatible with public metabolomics databases?

All identified acylcarnitines are annotated with HMDB or Lipid Maps IDs, allowing direct integration into tools like MetaboAnalyst or KEGG for pathway enrichment analysis.

What if my samples are from non-model organisms?

Our assays are species-agnostic, as acylcarnitine structures are conserved across mammals. We adapt extraction protocols for unique matrices (e.g., plant or insect samples) upon request.

Can I track changes in even-/odd-chain acylcarnitines separately?

Yes, our quantitative panel differentiates even-chain (e.g., C16) from odd-chain (e.g., C5) species, which may reflect distinct nutrient utilization (e.g., gut microbiota-derived metabolites).

How do you handle samples with hemolysis or lipemia?

Mild hemolysis/lipemia has minimal impact due to our SPE cleanup steps. However, severely compromised samples may require re-collection, as RBC lysis can artificially elevate certain species (e.g., C16:1).

Publications

References

  1. Zhang, Jingxian, et al. "Simultaneously quantifying hundreds of acylcarnitines in multiple biological matrices within ten minutes using ultrahigh-performance liquid-chromatography and tandem mass spectrometry." Journal of Pharmaceutical Analysis 14.1 (2024): 140-148.
  2. Giesbertz, Pieter, et al. "An LC-MS/MS method to quantify acylcarnitine species including isomeric and odd-numbered forms in plasma and tissues." Journal of lipid research 56.10 (2015): 2029-2039.
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