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Why Spermidine Matter

Spermidine is a small, highly polar polyamine. In real samples, that combination often makes measurement harder than expected: co-extractives can interfere, ionization can shift between matrices, and results may look "different" simply because the sample background is different. That's why many teams choose targeted LC-MS/MS for spermidine—especially when the readout will drive a decision.

Spermidine is also frequently interpreted in the context of related polyamines such as putrescine and spermine. Depending on your goal, a single-analyte measurement may be enough, or you may benefit from a polyamine panel to understand whether an intervention changes overall polyamine balance rather than spermidine alone.

What Problems This Service Helps You Solve

Confirm whether spermidine truly changes across experimental groups
Support treatment/control comparisons, dose–response studies, and time-course designs.
Reduce matrix-driven uncertainty
Apply matrix-aware preparation and QC so results remain comparable across sample types.
Choose the right level of interpretation
Start with spermidine-only for a focused question, or use a panel for pathway context.
Get outputs you can immediately use
Receive structured tables and QC documentation suitable for internal decision-making and reporting.

Precision Spermidine Quantification and Profiling Solutions

Creative Proteomics provides high-sensitivity, targeted analysis designed to overcome the specific chemical challenges of polyamine detection—such as high polarity and low molecular weight—ensuring your study yields definitive, high-integrity data.

Flexible Service Modules for Targeted Research

We offer adaptable testing protocols designed to align with your project's regulatory requirements and data depth needs:

Targeted Spermidine Absolute Quantification: Precise molar concentration determination using isotope-labeled internal standards.
Spermidine/Spermine Ratio Reporting: A critical assessment tool for monitoring cellular growth, senescence, and metabolic shifts.
Acetylated Flux Profiling: Simultaneous detection of N1-Acetylspermidine and N8-Acetylspermidine to track metabolic degradation and enzymatic activity.
Method Validation Services: Bespoke protocol development and validation following strict analytical guidelines for proprietary formulations.

Decision Matrix: Selecting the Right Analytical Scope

Use this guide to select the most informative service level for your study—ensuring you obtain the necessary data resolution without unnecessary complexity.

Your Research Goal	Recommended Choice	Why This Works	Typical Add-Ons
Confirm if spermidine changes between groups	Single-Analyte Spermidine Quantification	Focused, high-throughput answer when spermidine is your primary decision metric.	Absolute quantification; matrix-fit QC
Interpret pathway direction or conversion balance	Polyamine Panel	Providing context for Putrescine/Spermidine/Spermine reduces risk of misinterpretation.	Spermidine/Spermine ratio reporting
Track turnover and metabolic handling	Polyamine Panel + Derivatives	Acetylated forms add essential context when primary analyte changes are subtle.	N1 / N8-Acetylspermidine profiling
Work with complex or multiple matrices	Panel + Enhanced QC	Improves comparability and data confidence across diverse sample types.	Spike/recovery; matrix-effect checks
Link to functional readouts (e.g., autophagy)	Either (Scope dependent)	Single-analyte for focused correlation; panels to interpret broader systemic shifts.	Correlation-ready reporting format

Analytes We Can Quantify

Tier	Analytes	Notes
Default (Core)	Spermidine	Single-analyte service includes this only
Panel Core	Putrescine, Spermine	Included in polyamine panel
Optional Derivatives	N1-Acetylspermidine, N8-Acetylspermidine, N1-Acetylspermine	Added when turnover/routing context is needed
Custom Add-Ons	DAP and other pathway-related amines	Availability depends on matrix and study design

Advantages of Our Analysis Platform

High specificity in complex matrices using targeted LC-MS/MS with MRM transitions
Precision (repeatability): QC replicates typically achieve ≤10% CV (matrix-dependent)
Linearity: calibration curves typically achieve R² ≥ 0.99 across the working range
Sensitivity tuned to your matrix: LOD/LOQ optimized per sample type and prep strategy
Matrix-control options: spike/recovery and matrix-effect checks available when required

Our Spermidine Analysis Workflow

Spermidine LC-MS/MS analysis workflow

LC-MS/MS Platform for Spermidine Quantification

For reliable spermidine analysis in complex research matrices, we use a UHPLC–triple quadrupole LC-MS/MS setup optimized for targeted analysis.

Core Instruments (Most Common Setup)

Agilent 1290 UHPLC System
Agilent 6495C Triple Quadrupole LC/MS

Key Parameters For Spermidine Analysis

Parameter	TypicalSetting / Approach	Why ItMatters
Ionization	ESI (+)	Strong fit for polyamines
Acquisition	MRM (Multiple Reaction Monitoring)	High specificity and quantification confidence incomplex matrices
Target Mass Range	Optimized for low m/z analytes	Matches spermidine's small-molecule profile
LC Strategy	UHPLC separation tuned for polar compounds	Improves retention and reduces interferences
Optional Enhancement	Derivatization when appropriate	Can improve retention/response for challenging samples

Agilent 6495C Triple Quadrupole (Figure from Agilent)

Agilent 1290 UHPLC System (Figure from Agilent)

Sample Submission & Preparation Guidelines

Proper sample handling is critical to preserving the stability of biogenic amines. Use the following table as a baseline for your submission. If you are working with limited material, mixed matrices, or proprietary solvents, please consult our technical team to adapt the preparation and QC plan.

Sample type	Recommended amount	Container	Storage & shipping (general)	Notes
Cell pellet	≥ 1–5 million cells	Screw-cap tube	Freeze; ship cold/frozen	Avoid repeated freeze–thaw
Cell lysate	≥ 100–200 µL	Low-bind tube	Freeze; ship cold/frozen	Share protein conc. if available
Tissue	≥ 20–50 mg	Cryovial	Freeze; ship cold/frozen	Record wet weight
Culture media / supernatant	≥ 200–500 µL	Screw-cap tube	Freeze preferred	Note serum/medium type
Fermentation broth / microbial culture	≥ 0.5–1 mL	Screw-cap tube	Freeze preferred	Feasibility step recommended for challenging matrices
Plant tissue / extract	≥ 50 mg or ≥ 200 µL extract	Cryovial	Freeze; ship cold/frozen	Note solvent and extraction steps

Expert Handling Note: Polyamines like spermidine are sensitive to enzymatic degradation. We recommend immediate acidification or flash-freezing in liquid nitrogen post-collection to ensure the most accurate metabolic snapshot.

What You Receive: Actionable Deliverables

Executive Summary: A structured overview of analyte concentrations and significant findings.
Quantitative Data Suite: Detailed Excel/CSV tables featuring absolute concentrations, standard curves, and R2 values.
QC Documentation: Matrix spike recoveries, blank analysis, and internal standard performance to verify data integrity.
Technical Methodology: A clear description of instrumentation and preparation steps for internal reporting or regulatory dossiers.

LC-MS/MS MRM traces showing spermidine peak in sample and spike, absent in blank.

MRM chromatogram overlay showing spermidine peak specificity in blank, spike, and sample.

Calibration plot and residuals for spermidine LC-MS/MS quantification.

Spermidine calibration curve with residuals supporting linear, accurate absolute quantification.

Bar chart of spermidine recovery and matrix effects across sample types.

Recovery and matrix-effect summary across common matrices for robust spermidine quantification.

QC CV distribution for spermidine quantification with pass indicators.

QC precision plot showing reproducible spermidine quantification with acceptance threshold reference.

Research Applications for Spermidine and Metabolite Analysis

Autophagy & Cellular Stress Research

Relate intracellular spermidine shifts to stress-response readouts in non-clinical models.

Aging & Senescence Mechanism Work

Track spermidine dynamics across perturbations to support senescence-related hypotheses.

Immunometabolism & Immune Cell Function

Compare spermidine levels across activation states, treatments, or engineered conditions.

Microbiome–Host Metabolite Research

Measure spermidine in complex matrices to support microbe-derived metabolite studies.

Fermentation & Bioprocess Development

Monitor spermidine trends to compare strains, media, and process conditions.

Strain Engineering & Metabolic Optimization

Verify pathway edits by quantifying spermidine changes and optional derivatives.

Plant Physiology & Stress Response

Profile spermidine in tissues/extracts to support stress adaptation and regulation studies.

Nutritional Biochemistry & Food Research

Quantify spermidine in complex food-like matrices for formulation or process comparisons.

Q: What is the typical sensitivity for targeted spermidine LC-MS/MS analysis?

Our targeted assay achieves ultra-trace sensitivity, typically reaching Lower Limits of Quantitation (LLOQ) in the nanomolar (nM) or sub-ng/mL range. By utilizing the Agilent 6495C's iFunnel technology, we maximize ion transmission for small, polar molecules. This high sensitivity is essential for detecting spermidine in limited sample volumes, such as primary cell cultures, biopsies, or rare clinical specimens, where endogenous concentrations may be low.

Q: How do you address matrix suppression in complex biological samples?

To mitigate matrix effects—a common challenge for highly polar polyamines—we employ Isotope-Dilution Mass Spectrometry (IDMS). By adding stable isotope-labeled internal standards (¹³C or ¹⁵N-Spermidine) to every sample during the extraction phase, we can precisely correct for ionization suppression or enhancement caused by the sample background. This ensures that the reported concentrations are absolute and comparable across diverse matrices, such as serum, brain tissue, and fermentation broths.

Q: What is the best way to normalize spermidine concentration data?

Data normalization is critical for biological interpretation. We provide multiple normalization strategies depending on your matrix:

Cell Pellets/Lysates: Normalized by total protein concentration (BCA/Bradford) or total cell count.
Tissues: Normalized by milligrams of wet weight or dry weight (lyophilized).
Biofluids: Normalized by volume or creatinine levels (for urine).

Spermidine/Spermine ratios are also reported as a dimensionless biomarker for metabolic homeostasis.

Q: How does sample degradation affect spermidine quantification?

Spermidine is highly susceptible to enzymatic degradation by endogenous polyamine oxidases (PAO). If samples are not stabilized immediately, spermidine can rapidly convert into putrescine or degrade further, leading to underestimation. We recommend immediate flash-freezing in liquid nitrogen or stabilization with acidic buffers (e.g., Perchloric acid or HCl) post-collection to "lock" the metabolic state and ensure data integrity.

Q: Is spermidine a reliable biomarker for monitoring autophagy in vivo?

Spermidine is considered a key endogenous inducer and biomarker of autophagy. Research shows that intracellular spermidine levels decrease with age, and its replenishment correlates with the upregulation of autophagic flux (e.g., LC3-II/LC3-I ratios). Quantifying spermidine via LC-MS/MS provides a direct metabolic readout of the autophagic capacity in longevity studies, drug efficacy trials, and disease-state monitoring.

Q: Why choose targeted LC-MS/MS over untargeted metabolomics for polyamines?

Untargeted metabolomics often fails to detect spermidine due to its high polarity and poor retention on standard C18 columns. Our targeted LC-MS/MS service utilizes optimized stationary phases and MRM transitions specifically tuned for small polyamines. This provides superior specificity, a wider dynamic range, and a much lower LLOQ than discovery-based methods, making it the gold standard for clinical and regulatory-grade quantification.

Polyamine metabolism impacts T cell dysfunction in the oral mucosa of people living with HIV

Mahalingam, S. S., Jayaraman, S., et al.

Journal: Nature Communications

Year: 2023

DOI: https://doi.org/10.1038/s41467-023-36163-2

Nutritional supplement containing a nuclear fraction of bovine thymus gland increases the circulating levels of spermidine

Surzenko, N., et al.

Journal: PLoS One

Year: 2025

DOI: https://doi.org/10.1371/journal.pone.0331813

Spermidine Quantification Service: Targeted LC-MS/MS