Thiols Analysis Service

Get reliable thiol quantification without oxidation artifacts.

Creative Proteomics stabilizes, derivatizes, and quantifies biothiols and volatile thiols across complex matrices—so you can track redox status, process consistency, and product quality with confidence.

  • We solve: unstable thiols, matrix interferences, low-abundance detection
  • You get: verified GSH/GSSG ratios, comprehensive biothiol panels, volatile thiol fingerprints
  • Proof in numbers: LLOQ down to 0.5–10 nM (biothiols) and sub-ng/L (volatile thiols); linearity r² ≥ 0.995 over ≥4 orders; precision CV ≤ 10%
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What You Will Receive

  • Quantified Thiol Levels: Concentrations of GSH, Cys, 3MH, and others, reported in nM or ng/L.
  • Redox Ratios & Metrics: GSH/GSSG, Cys/Cystine, Hcy ratios, and optional redox potential (Eₕ).
  • QC-Verified Results: Includes calibration curves, recovery, CVs, blanks, and QC plots.
  • Raw & Processed Data: Delivered in .raw / .wiff, plus .mzML and .csv summary files.
  • What We Provide
  • Advantages
  • Technology Platform
  • Sample Requirement
  • Demo
  • FAQs

What Are Thiols and Why Analyze Them?

Thiols (–SH compounds) are reactive sulfur-containing molecules critical for redox balance, antioxidant defense, and aroma profiles in biological and industrial systems. Key examples include glutathione (GSH) and cysteine in cellular redox buffering, as well as 3MH and 2-furfurylthiol in fermented beverages.

However, their high reactivity also makes them analytically unstable. Without rapid stabilization, thiols oxidize or degrade, leading to inaccurate quantification and misleading data. Conventional workflows often capture artifacts rather than true in-sample concentrations.

Targeted thiols analysis—with appropriate derivatization, internal standards, and matrix-specific methods—is essential for reliable results. Whether assessing GSH/GSSG ratios, profiling volatile sulfur compounds, or tracking low-abundance thiols in bioprocesses, accurate thiol data provides actionable insights into product quality, biological state, and process performance.

Thiols Analysis Services Offered by Creative Proteomics

  • Targeted Biothiols Panel (quantitative): Cys, Cys-Gly, Hcy, γ-Glu-Cys, GSH/GSSG, CoA-SH, NAC, Cystine, Homocystine
  • Extended Sulfur Metabolites (semi-quant/quant): Cysteamine, 3-mercaptopropionic acid, 2-mercaptoethanol, penicillamine
  • Volatile Thiols Fingerprinting (targeted GC–MS/MS): 2-furfurylthiol, 4MMP, 3MH, 3MHA, MFT, benzyl mercaptan, ethanethiol
  • Protein Thiol/Disulfide State (mapping): Differential alkylation and peptide-level LC–MS for relative thiol occupancy
  • Redox Metrics: GSH/GSSG, Cys/Cystine, Hcy/Homocystine ratios and Nernst-based redox potential (optional)
  • Stability & Stress Studies: Thiol loss kinetics under process conditions; antioxidant/additive screening

Thiols Analysis Targets & Compound Panels

PanelRepresentative AnalytesTypical ApplicationTypical LLOQ*
Core Biothiols (LC–MS/MS)- Glutathione (GSH)
- Oxidized Glutathione (GSSG)
- Cysteine (Cys)
- Cystine
- Homocysteine (Hcy)
- Homocystine
Redox balance, oxidative stress, metabolic studies0.5–10 nM
Extended Biothiols (LC–MS/MS)- γ-Glutamylcysteine
- Cysteinylglycine
- N-acetylcysteine (NAC
- Coenzyme A (CoA-SH)
- Cysteamine
- 3-Mercaptopropionic acid
- 2-Mercaptoethanol
- Penicillamine
Drug metabolism, cell signaling, thiol scavenger profiling1–20 nM
Volatile Thiols (GC–MS/MS)- 3-Mercaptohexanol (3MH)
- 3-Mercaptohexyl acetate (3MHA)
- 4-Mercapto-4-methylpentan-2-one (4MMP)
- 2-Furfurylthiol (FFT)
- 2-Methyl-3-furanthiol (MFT)
- Benzyl mercaptan
- Methanethiol
- Ethanethiol
Flavor/aroma profiling in wine, beer, fermented products0.2–2 ng/L
Thiol Ratios & Redox Metrics- GSH/GSSG ratio
- Cys/Cystine ratio
- Hcy/Homocystine ratio
- Redox potential (Eₕ, optional)
Redox state monitoring, stress response profiling

Advantages of Our Thiols Analysis Services

  • Sensitivity & Range: LLOQ 0.5–10 nM (biothiols) and 0.2–2 ng/L (volatiles); dynamic range ≥ 10⁴ with weighted calibration (r² ≥ 0.995).
  • Accuracy & Precision: Isotope dilution recovery 85–115%; intra-run precision CV ≤ 10%, inter-run ≤ 15% across matrices.
  • True-state Capture: Immediate NEM/IAM capping prevents >90% of artifactual GSH oxidation observed without stabilization.
  • Carryover Control: Post-run and needle wash validated to <0.1% for high-abundance thiols.
  • Trace Volatiles: SPME enrichment + MS/MS achieves signal-to-noise ≥ 10 at sub-ng/L with matrix-matched standards.
  • QC Transparency: Blanks, duplicates, matrix spikes, and low/medium/high QCs bracket every batch; full QA plots provided.

Workflow for Thiols Analysis Service

1. Consult & Panel Design: Define targets, matrices, expected ranges, stabilization strategy (NEM/IAM/mBBr).

2. Method Setup: Choose LC–MS/MS, GC–MS/MS, or hybrid; assign internal standards and calibration range.

3. Sample Stabilization: Provide pre-loaded tubes or on-receipt alkylation; confirm pH/temperature controls.

4. Extraction & Cleanup: Protein precipitation or SPE; HILIC/RP selection to minimize matrix effects.

5. Acquisition: MRM-based quant with bracketed QCs, blanks, and matrix spikes; volatility-optimized SPME for aroma thiols.

6. Quantification & QA: Weighted regression, back-calculation checks, recovery and precision acceptance, carryover review.

7. Reporting: Concentrations (with units), key ratios (e.g., GSH/GSSG), method metadata, calibration/QC plots, and raw files.

Thiols Analysis Workflow

Technology Platform for Thiols Analysis Service

LC–MS/MS for Biothiols

Widely used for quantifying GSH, GSSG, cysteine, homocysteine and related thiols in biological matrices (plasma, tissues, cell lysates).

Instrumentation:

UHPLC: Agilent 1290 Infinity II or Waters ACQUITY UPLC

MS/MS: AB Sciex Triple Quad 6500+ or Waters Xevo TQ-S micro

Chromatography:

Column: 2.1 × 100 mm, 1.7 μm C18 or HILIC

Derivatization & Stabilization:

N-ethylmaleimide (NEM) or iodoacetamide (IAM) to prevent oxidation prior to extraction

MS Parameters:

Ion source: ESI positive mode

Quantification:

Internal standards: ¹³C₂-Gly, d₅-GSH, d₄-Cys, etc.

Calibration: 1/x or 1/x² weighting

LOQ: Typically 0.5–10 nM

GC–MS/MS for Volatile Thiols

Used to detect ultra-trace levels of aroma-active thiols in beverages, fermentation broths, and flavor samples.

Instrumentation:

GC: Agilent 7890B

MS/MS: Agilent 7010B Triple Quadrupole

Ionization: EI or NCI (depending on derivatization)

Sample Preparation:

Preconcentration via SPME (DVB/CAR/PDMS fiber)

Salt and pH adjustment to optimize recovery

Derivatization (if applicable):

Pentafluorobenzyl bromide (PFB-Br) for enhanced volatility and MS response

Quantification:

Stable-isotope dilution (e.g., d₄-3MH, d₂-4MMP)

Method-specific MRM transitions

LOQ: Typically 0.2–2 ng/L, matrix-dependent

SCIEX Triple Quad™ 6500+

SCIEX Triple Quad™ 6500+

Agilent 5977B GC/MSD

Agilent 5977B GC/MSD

Waters ACQUITY UPLC System

Waters ACQUITY UPLC System

Thermo Fisher Q Exactive

Thermo Fisher Q Exactive

Sample Requirements for Thiols Analysis Service

Sample TypeMinimum AmountContainer & PreservativeHandling & StorageNotes
Plasma / Serum≥100 μLEDTA or heparin tube; transfer to amber microtube pre-loaded with NEM (final ~10 mM)Chill on ice immediately; snap-freeze; ship on dry iceAvoid hemolysis; ≤1 freeze-thaw
Whole Blood (for rapid GSH/GSSG)200 μLTube pre-loaded with NEM; gentle inversionProcess or freeze promptly; dry ice shippingRecord anticoagulant type
Urine≥500 μLAmber tube; optional NEM stabilizationFreeze at −80 °C; dry ice shippingProvide creatinine for normalization (optional)
Cell Pellets≥1×10⁶ cells or ≥10 mg wet massPre-cooled tube; add alkylating PCA buffer on receiptKeep cold; snap-freezeSupply cell count/protein if available
Tissue≥20 mgCryovial; no buffer; dry iceSnap-freeze post-collection; store −80 °CDo not thaw before shipment
Fermentation / Media≥1 mLPolypropylene; optional filtration 0.22 μm; NEM if biothiols targetedFreeze; dry ice shippingRecord medium composition
Beverages (volatile thiols)≥10 mLAmber headspace vials filled to minimize headspaceChill; ship cold or on ice packs; avoid lightProvide pH and ethanol (% v/v)
Formulations≥1 mL or ≥100 mgOriginal container plus aliquot in amber tubeProtect from light; ship coldInclude excipient list for matrix match

Demo Results

Calibration curve and QC plot showing linear response and recovery accuracy for thiol analysis using LC–MS/MS.

Calibration curve and QC validation for thiol quantification. The method demonstrates excellent linearity (R² > 0.995) and recovery within ±15% QC limits across low, mid, and high concentrations.

Heatmap displaying volatile thiol concentration differences (in ng/L) among multiple samples, used for flavor profiling.

Heatmap of volatile thiol concentrations across multiple samples. Distinct profiles of 3MH, 4MMP, MFT, and related compounds highlight sample-to-sample variation in aroma-active sulfur compounds.

Chromatogram showing GC–MS peak separation of volatile thiols such as 3MH and 4MMP by retention time and ion intensity.

Simulated GC–MS XIC of six key volatile thiols with distinct retention times, illustrating effective chromatographic separation for 3MH, 3MHA, 4MMP, and others.

FAQ of Thiols Analysis Service

How is thiol stability maintained during sample transport and processing?

Alkylating agents such as NEM or IAM are applied immediately upon sample collection or receipt. This prevents disulfide formation and ensures true-state quantification of reduced thiols.

What controls are included to ensure data quality and comparability across batches?

All runs are bracketed with internal standards, matrix-matched calibration curves, and multi-level QCs (low/mid/high). Recovery, CVs, and back-calculation are reported per analyte.

Can the method distinguish between endogenous thiols and exogenous thiol-containing additives?

Yes. Targeted MRM transitions and isotope dilution quantification enable compound-specific resolution, even in the presence of formulation excipients or derivatized analogs.

Is the method validated for complex matrices like fermentation broth or high-protein samples?

Yes. Matrix-specific cleanup (e.g., SPE or precipitation) and method adjustments are implemented to handle high protein, sugar, or salt content without compromising sensitivity or selectivity.

How are thiol-related redox metrics (e.g., GSH/GSSG) calculated and interpreted?

Molar concentrations are directly measured and used to compute relevant ratios. Optional redox potential (Eₕ) calculations can be provided using the Nernst equation.

Are volatile thiols quantified in absolute or relative terms?

Absolute quantification (ng/L) is performed using matrix-matched standards and deuterated internal references. This enables batch comparability and aroma intensity ranking.

Can this platform support thiol fingerprinting for batch release or raw material QC?

Yes. Volatile thiol profiles or total biothiol content can be used as stability or identity markers for QC applications in manufacturing pipelines.

Is differential alkylation for thiol/disulfide mapping supported at the peptide level?

Yes. Peptide-level LC–MS/MS analysis with differential labeling can identify site-specific thiol modifications or disulfide scrambling in proteins under redox stress.

What's the typical carryover risk in high-abundance thiol samples?

Carryover is routinely tested and controlled below 0.1% using post-run solvent blanks and dedicated needle wash cycles.

Can custom thiol panels be aligned with previous in-house or published workflows?

Yes. Custom panels can replicate legacy methods or published MRM transitions. Cross-platform method harmonization and comparison studies can be provided on request.

Learn about other Q&A about proteomics technology.

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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.

Metabolomics Sample Submission Guidelines
* For Research Use Only. Not for use in diagnostic procedures.
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