Branched-Chain Amino Acids (BCAAs) Analysis Service

What Are Branched-Chain Amino Acids (BCAAs)?

BCAAs—L-leucine, L-isoleucine, L-valine—are essential amino acids with branched aliphatic side chains. They are central to nitrogen handling, carbon routing, and anabolic signaling. In applied settings (food, feed, fermentation, and discovery research), accurate BCAA quantification supports formulation control, process stability, and mechanistic insight.

What Problems We Solve for Our Clients

Quantify BCAA levels with confidence

Research-grade LC–MS/MS for precise leucine, isoleucine, valine concentrations to answer "how much is present?"

Compare groups, time points, and interventions

Detect meaningful differences across control vs. treatment, baseline vs. post-intervention, or longitudinal designs.

Gain pathway insight

Integrate BCAAs with branched-chain keto acids (BCKAs) and acylcarnitines to map transamination and downstream catabolism.

Evaluate formulations and prototypes (R&D)

Check free BCAA levels in development samples to guide composition tuning toward target ranges.

Assess stability and handling effects

Determine how storage, processing, or protocol changes impact BCAA retention to refine SOPs.

Support longitudinal and trend analysis

Track BCAA changes over time to reveal dynamic patterns, assess intervention effects, and strengthen data interpretation.

What We Offer — Branched-Chain Amino Acids Analysis from Creative Proteomics

• Targeted BCAA quantification (primary panel): Absolute concentrations of leucine, isoleucine, valine using stable-isotope internal standards.
• BCKA add-on panel: α-ketoisocaproate (KIC), α-keto-β-methylvalerate (KMV), α-ketoisovalerate (KIV).
• Acylcarnitine add-on: Isovaleryl-, isobutyryl-, and 2-methylbutyrylcarnitine for catabolic read-through.
• Extended amino-acid bundle: ≥20 amino acids for nutritional and process context.
• Isotope-tracing (research): ^13C/^15N-BCAA tracers to examine pathway flux in cells, tissues, or fermenters.
• Cross-platform confirmation (as needed): GC–MS on derivatized analytes for challenging matrices.
• Data analysis options: Time-course trend plots, PCA across groups, correlation with your process variables (e.g., pH, DO, glucose).

Full Panel of Branched-Chain Amino Acids and Related Metabolites

Why Choose Our Branched‑Chain Amino Acids Analysis: Key Advantages

• Validated Sensitivity for Multiple Matrices: Matrix-qualified LOQs as low as 5–50 nM in biofluids and 0.5–5 µM in food/feed/tissue samples, ensuring detection even in low-abundance scenarios.
• Wide Quantification Range: Linear response across 3–4 orders of magnitude (r² ≥ 0.995) to accommodate both trace-level and high-concentration samples without re-analysis.
• High Precision and Accuracy: Intra- and inter-batch precision typically RSD ≤ 10%; spike-recovery results within 85–115%, delivering reproducible data for confident comparisons.
• Matrix Effect Control: Isotope-dilution quantification combined with matrix-matched calibration minimizes ion suppression/enhancement for true concentration values.
• Batch-to-Batch Consistency: Calibrator bracketing and QC monitoring throughout the run safeguard consistency across large sample sets and multi-batch studies.
• Traceable, Auditable Data: Each reported value is linked to its chromatogram, integration file, and QC record, enabling transparent verification for reports, publications, or audits.

Technical Specifications You Can Rely On

Primary platform: UHPLC–MS/MS (MRM)

• UHPLC systems (typical): Thermo Vanquish or Waters ACQUITY UPLC I-Class.
• Triple-quad MS (typical): Thermo TSQ Altis, SCIEX QTRAP 6500+, or Waters Xevo TQ-XS.
• Ionization: ESI (+), MRM transitions optimized per analyte; dwell times adjusted for peak fidelity.
• Chromatography options:
  • HILIC: BEH Amide, 2.1 × 100 mm, 1.7 µm; high-organic starts for sharp early peaks.
  • RP with derivatization: C18, 2.1 × 100 mm, 1.6–1.8 µm using AQC (AccQ-Tag) or PITC chemistry to improve retention and response.
• Mobile phases: Volatile buffers (ammonium formate/acetate) with formic acid; aqueous/organic gradients tuned per matrix.
• Calibration & QC: ≥7 levels matrix-matched; low/mid/high QCs within-batch; blank and solvent checks every sequence segment.
• Internal standards: ¹³C6-Leu, ¹³C6-Ile, ¹³C5-Val matched to each native BCAA.

Complementary confirmation (as needed): GC–MS

• System (typical): Agilent 7890 GC with 5977B MSD.
• Column: Low-bleed phenyl-arylene (e.g., DB-5ms, 30 m × 0.25 mm × 0.25 µm).
• Derivatization: TBDMS (MTBSTFA + catalyst) or TMS (BSTFA + catalyst), selected per matrix.
• Acquisition: EI mode; temperature gradient optimized for full baseline resolution of BCAA derivatives.

Waters ACQUITY UPLC System (Figure from Waters)

SCIEX Triple Quad™ 6500+ (Figure from Sciex)

TSQ Altis Triple Quadrupole MS (Figure from Thermo Scientific)

Agilent 7890B-5977B (Figure from Agilent)

How Our BCAA Analysis Works — Step-by-Step Process

Accepted Sample Types and Collection Instructions for BCAA Quantification

Deliverables: What You Get from Our Branched‑Chain Amino Acids Assay

• BCAA concentration data（L-Leucine, L-Isoleucine, L-Valine）
• Optional related metabolites（BCKAs, acylcarnitines, other amino acids, if selected）
• QC summary（calibration curve, accuracy, precision）
• Representative chromatograms
• Data files：Excel/CSV & PDF report
• Optional charts：trend plots, group comparisons

LC–MS/MS chromatograms showing baseline-separated peaks for L-Leucine, L-Isoleucine, and L-Valine with internal standard control.

Representative mass spectrum of a branched-chain amino acid, highlighting characteristic fragment ions and isotope-dilution quantification.