Ss31 Coa
Evaluating an SS-31 Certificate of Analysis (COA) is essential for validating chemical purity, identity, and batch consistency in mitochondrial research applications. PX1 Research provides fully documented, third-party verified SS-31 for high-precision laboratory and in vitro investigations.
Quick answer
Evaluating an SS-31 Certificate of Analysis (COA) is essential for validating chemical purity, identity, and batch consistency in mitochondrial research applications. PX1 Research provides fully documented, third-party verified SS-31 for high-precision laboratory and in vitro investigations.
Key takeaways
- An [SS-31](/research-peptides/ss-31) [Certificate of Analysis](/research-peptides/what-is-a-coa-for-peptides) (COA) is an official analytical document provided by an independent ISO 17025 accredited laboratory that verifies the precise purity, exact molecular weight, sequence identity, and endotoxin thresholds of a specific lot of synthesized SS-31 peptide.
- [SS-31](/research-peptides/ss-31), also designated as Elamipretide or Szeto-Schiller 31, is a tetrapeptide with the chemical sequence D-Arg-2',6'-Dmt-Lys-Phe-NH2.
- Preclinical studies indicate that the primary mechanism of action for [SS-31](/research-peptides/ss-31) centers on its high-affinity electrostatic and hydrophobic interactions with cardiolipin, a unique tetra-acyl phospholipid localized exclusively to the inner mitochondrial membrane.
- Laboratory investigations using isolated mitochondria, cultured cell lines, and animal models have widely utilized [SS-31](/research-peptides/ss-31) to explore cardiolipin protection and mitochondrial repair mechanisms.
Understanding the SS-31 Certificate of Analysis (COA)
An SS-31 Certificate of Analysis (COA) is an official analytical document provided by an independent ISO 17025 accredited laboratory that verifies the precise purity, exact molecular weight, sequence identity, and endotoxin thresholds of a specific lot of synthesized SS-31 peptide. It serves as the primary standard for quality assurance in quantitative in vitro and preclinical research.
For investigators studying bioenergetics, mitochondrial membrane integrity, and oxidative stress pathways, obtaining a lot-specific COA is critical. Variations in peptide synthesis, residual trifluoroacetic acid (TFA) salts, micro-aggregates, or bacterial endotoxins can drastically alter cellular assays, skew mitochondrial respiration assays, or induce non-specific cytotoxic responses. A comprehensive COA eliminates these experimental variables by confirming that the supplied SS-31 research compound meets rigorous chemical standards prior to reconstitution.
Chemical Identity and Structural Overview of SS-31
SS-31, also designated as Elamipretide or Szeto-Schiller 31, is a tetrapeptide with the chemical sequence D-Arg-2',6'-Dmt-Lys-Phe-NH2. It possesses a molecular formula of C32H49N9O5 and a theoretical monoisotopic mass of approximately 639.39 g/mol. Characterized by an alternating aromatic-cationic structural motif, SS-31 was specifically designed to readily penetrate cell membranes and selectively accumulate within the inner mitochondrial membrane (IMM).
Unlike conventional lipophilic cations that rely on the mitochondrial membrane potential (ΔΨm) for accumulation, the cellular uptake of SS-31 is independent of membrane potential. This property allows researchers to investigate dysfunctional or depolarized mitochondria in preclinical models without compromising peptide delivery. High-grade synthetic SS-31 offered across our full catalog of research peptides undergoes stringent solid-phase peptide synthesis (SPPS) to ensure correct stereochemistry and structural fidelity.
Mitochondrial Targeting Mechanism: Cardiolipin Interactions
Preclinical studies indicate that the primary mechanism of action for SS-31 centers on its high-affinity electrostatic and hydrophobic interactions with cardiolipin, a unique tetra-acyl phospholipid localized exclusively to the inner mitochondrial membrane. Cardiolipin is integral to organizing electron transport chain (ETC) complexes into functional supercomplexes (respirasomes) and maintaining optimal cristae curvature.
Under conditions of oxidative stress, cardiolipin undergoes peroxidation, which disrupts supercomplex assembly, impairs ATP synthase efficiency, and leads to excessive production of reactive oxygen species (ROS). In vitro data demonstrate that SS-31 selectively binds to cardiolipin via its dimethyltyrosine (Dmt) and basic amino acid residues. By stabilizing cardiolipin structure, SS-31 prevents pathological phase transitions, optimizes electron transfer across Complexes I–IV, inhibits cytochrome c release, and reduces electron leakage without altering baseline respiration rates in healthy control models.
Preclinical Research Findings in Bioenergetic Models
Laboratory investigations using isolated mitochondria, cultured cell lines, and animal models have widely utilized SS-31 to explore cardiolipin protection and mitochondrial repair mechanisms. Research in rodent models of ischemia-reperfusion injury highlights that SS-31 administration prior to or during reperfusion reduces ROS generation, preserves ATP levels, and attenuates apoptotic pathway activation.
Furthermore, preclinical models examining metabolic decline, neurodegeneration, and age-related muscular fatigue indicate that SS-31 targeted therapy preserves mitochondrial structural integrity across diverse tissue types. Researchers utilizing our mitochondrial research peptides frequently analyze SS-31 alongside other mitochondrial-derived peptides to map out overlapping pathways regulating cellular homeostasis and metabolic efficiency.
Decoding the SS-31 COA: Key Analytical Metrics
A reliable SS-31 COA must contain several critical analytical parameters that verify the physical and chemical state of the lyophilized powder. When reviewing documentation, research teams should scrutinize the following sections: Analytical Method (RP-HPLC/LC-MS), Purity Percentage, Mass Spectrometry Peak Confirmation, Counterion Analysis, Moisture Content, and Endotoxin Quantification.
At PX1 Research, every batch of SS-31 is subjected to stringent testing protocols. A pass status requires a minimum purity threshold of ≥98% as determined by high-resolution Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC), with complete structural confirmation via Liquid Chromatography-Mass Spectrometry (LC-MS). Batch records are accessible through our dedicated research library hub to assist laboratory technicians in data validation.
RP-HPLC Analysis and Mass Spectrometry Verification
RP-HPLC is the gold standard method for determining the chemical purity of synthetic peptides. The technique separates the primary peptide sequence from synthesis side-products, deletion sequences, and truncated fragments based on hydrophobicity. The resulting chromatogram displays a single, dominant retention peak corresponding to SS-31, with area-under-the-curve (AUC) integration demonstrating ≥98% relative area.
Complementing HPLC, LC-MS analysis confirms the exact molecular mass of the peptide fragment. For SS-31, mass spectrometry verifies the observed mass-to-charge ratio (m/z) matches the theoretical molecular weight of the protonated ion ([M+H]+). This guarantees that the sample contains the intended sequence rather than an isobaric variant or incorrect sequence orientation.
Endotoxin Content and Sterility Requirements in Research Applications
Bacterial endotoxins (lipopolysaccharides) are common contaminants in peptide synthesis and handling. In cell culture assays and live animal tissue models, elevated endotoxin levels trigger severe inflammatory responses through Toll-like receptor 4 (TLR4) activation, confounding experimental results and causing non-specific cell mortality.
To prevent artifactual data, PX1 Research subjects all lot runs to Limulus Amebocyte Lysate (LAL) testing to ensure endotoxin limits remain strictly below 0.01 EU/mg. Combined with sterile manufacturing controls in GMP-compliant, USA-based facilities, research teams receive compounds suitable for sensitive primary cell cultures and delicate metabolic studies.
Comparative Analysis: SS-31 vs. Other Mitochondrial Compounds
In mitochondrial bioenergetics research, SS-31 is frequently compared with mitochondrial-derived peptides (MDPs) such as MOTS-c peptide and Humanin research compound. While SS-31 works via direct physical interaction with cardiolipin on the inner mitochondrial membrane to optimize electron flow, MOTS-c operates primarily as a nuclear signaling peptide that regulates metabolic transcription and systemic insulin sensitivity.
Similarly, Humanin functions largely as a cytoprotective factor that binds IGFBP-3 and suppresses Bax-mediated apoptosis under cellular stress. Researchers conducting comparative studies often utilize these peptides in tandem to differentiate direct membrane-level bioenergetic restoration (SS-31) from gene-regulatory metabolic signaling (MOTS-c and Humanin). For bulk exploratory programs, researchers can set up customized orders via our wholesale research portal.
Handling, Reconstitution, and Storage Protocols for Laboratory Use
Lyophilized SS-31 should be stored at -20°C or -80°C upon receipt to maintain long-term stability. The dry powder is highly stable when protected from moisture, light, and ambient temperature fluctuations. Prior to opening the vial, allow it to equilibrate to room temperature to prevent atmospheric moisture condensation on the lyophilized cake.
Reconstitution should be performed under a sterile laminar flow hood using laboratory-grade solvents such as Bacteriostatic Water, Sterile Saline (0.9% NaCl), or Ultra-Pure Water depending on the downstream assay requirements. SS-31 exhibits high solubility in aqueous buffer solutions due to its hydrophilic cationic residues. Reconstituted stock solutions should be aliquoted into single-use polypropylene microcentrifuge tubes and frozen at -80°C to avoid repeated freeze-thaw cycles that can induce peptide degradation.
PX1 Research Quality Assurance and USA Manufacturing
PX1 Research sets the industry standard for peptide quality control by manufacturing all compounds exclusively in USA-based, GMP-compliant facilities. Every product batch is independently tested by accredited third-party ISO 17025 laboratories, ensuring uncompromised purity and transparent analytical reporting.
Orders placed Monday through Friday ship same-day from our primary distribution hubs in California and Arizona. Each shipment includes a comprehensive, lot-matched Certificate of Analysis detailing HPLC retention curves, mass spectrometry readings, and endotoxin assays, giving researchers full confidence in their experimental inputs.
Frequently Asked Questions
What is an SS-31 COA?
An SS-31 COA (Certificate of Analysis) is a third-party analytical document detailing the purity (HPLC), mass identity (LC-MS), sequence verification, and endotoxin levels of a specific lot of SS-31 research peptide.
What purity level should a research-grade SS-31 COA show?
High-precision laboratory research requires an SS-31 purity of ≥98% by RP-HPLC AUC measurement, with mass spectrometry confirming the correct molecular mass.
How does SS-31 differ from MOTS-c in research models?
SS-31 directly binds cardiolipin in the inner mitochondrial membrane to optimize supercomplex structure and reduce ROS, whereas MOTS-c functions as a nuclear-encoded metabolic regulator affecting gene expression.
What is the acceptable endotoxin limit on an SS-31 COA?
For sensitive cell culture and in vivo rodent models, endotoxin levels should ideally measure below 0.01 EU/mg to prevent non-specific immune activation.
How should lyophilized SS-31 be stored upon arrival?
Lyophilized SS-31 powder should be stored desiccated at -20°C or -80°C, protected from light and heat, to ensure chemical stability for extended research timelines.
What solvents are suitable for reconstituting SS-31?
SS-31 is highly water-soluble and can be reconstituted using sterile water for injection, bacteriostatic water, or phosphate-buffered saline (PBS) under sterile laboratory conditions.
Where is PX1 Research SS-31 manufactured and tested?
PX1 Research peptides are manufactured in USA-based, GMP-compliant facilities and tested by independent ISO 17025 accredited laboratories.
Can SS-31 be used for human consumption or therapeutic treatment?
No. SS-31 provided by PX1 Research is strictly designated for laboratory research, in vitro studies, and preclinical scientific investigation only.
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