| HS Code | 523708 |
| Product Name | Glycyl-L-Histidyl-L-Lysine |
| Chemical Name | L-Glycyl-L-histidyl-L-lysine |
| Cas Number | 49557-75-7 |
| Molecular Formula | C14H24N6O4 |
| Molecular Weight | 340.38 g/mol |
| Sequence | Gly-His-Lys (GHK) |
| Iupac Name | (2S)-6-amino-2-[[(2S)-2-[[2-aminoacetyl]amino]-3-(1H-imidazol-4-yl)propanoyl]amino]hexanoic acid |
| Synonyms | GHK; Gly-His-Lys; L-Glycyl-L-histidyl-L-lysine |
| Appearance | White to off-white crystalline powder |
| Solubility | Soluble in water, DMSO, and aqueous buffers |
| Purity | ≥98% by HPLC |
| Storage Conditions | Store at -20°C, desiccated, protected from light |
| Smiles | C1=NC(=CN1)CC(C(=O)NC(CCCCN)C(=O)O)NC(=O)CN |
As an accredited Glycyl-L-Histidyl-L-Lysine factory, we enforce strict quality protocols—every batch undergoes rigorous testing to ensure consistent efficacy and safety standards.
| Packing | Glycyl-L-Histidyl-L-Lysine supplied as lyophilized powder in a sealed glass vial, 10 mg per vial. |
| Container Loading (20′ FCL) | Glycyl-L-Histidyl-L-Lysine is loaded as a 20-foot full container in sealed, moisture-proof drums, secured on pallets, with temperature control to ensure stability. |
| Shipping | Glycyl-L-Histidyl-L-Lysine (GHK) is shipped as a lyophilized peptide powder to ensure stability. Shipments are dispatched in insulated containers with ice packs, maintaining cool conditions during transit. Handle with care, store at -20°C upon receipt, and keep away from moisture, heat, and prolonged exposure to light. |
| Storage | Store Glycyl-L-Histidyl-L-Lysine (GHK) as a lyophilized powder at -20°C, protected from light and moisture. Keep the container tightly sealed and desiccated. Avoid repeated freeze-thaw cycles; aliquot solutions before freezing. Under these conditions, the peptide remains stable for extended periods. |
| Shelf Life | Shelf life is approximately 2 years when stored lyophilized at -20°C; after reconstitution, it remains stable for up to 1 month refrigerated. |
Post-emulsification addition of Glycyl-L-Histidyl-L-Lysine into high-water-phase o/w emulsions is performed at 38–42 °C with 150–250 rpm paddle agitation rather than rotor-stator shear, because rotor-stator dispersers operating at 3,000–8,000 rpm generate localized fluid temperatures above 60 °C near the rotor gap and reduce peptide recovery as measured by HPLC at 214 nm. The final leave-on emulsion typically carries 0.01–0.10 wt% GHK, prepared from a 1.0 wt% aqueous stock solution pre-filtered through 0.22 µm PVDF membrane. The formulation is buffered to pH 5.0–6.0 with 10–20 mM citrate or lactate buffer; phosphate buffer levels above 20 mM are avoided in lamellar gel networks containing fatty alcohols above 3.0 wt% because gel structure collapse reduces batch-to-batch viscosity repeatability. Terminal finished goods in this application class include o/w creams, lotions, gel-creams, and airless pump emulsions. Regulatory compliance for these finished goods falls under Regulation (EC) No 1223/2009 with a product information file prepared according to Article 10, manufacturing under ISO 22716, and microbial quality anchored to ISO 17516:2014 with preservation challenge testing according to ISO 11930:2019 criterion A. Production-scale failure modes observed on filling lines include viscosity drift when the peptide stock is dosed before full cooling of the batch, and air entrapment in formulations above 40,000 mPa·s when airless pumps are filled without vacuum deaeration.
Stability protocols for leave-on emulsions are typically supported by storage at 25 °C/60% RH for 24 months and accelerated storage at 40 °C/75% RH for 12 weeks, with pH, HPLC purity, and visual phase separation measured at defined intervals. Opaque laminated tubes or airless packages with limonene-free fragrances are preferred because the histidine residue is oxidation-prone under high oxygen headspace. The tripeptide is not introduced into the hot oil phase above 80 °C, and addition during the neutralization step of a soap-based emulsion is specifically avoided because rapid pH swing through 8.0–9.0 causes peptide degradation and uncontrolled viscosity reduction.
In transparent aqueous serums filled into 10–30 mL dropper bottles or 2–5 mL single-dose ampoules, haze formation is not a simple solubility failure of GHK in water but a buffer-induced aggregation event when multivalent electrolytes interact with the histidine side chain. Formulators using deionized water with conductivity below 2 µS/cm and no added sodium chloride can hold haze below 5 NTU after 12 weeks at 40 °C, while the same formula with 50 mM sodium chloride or magnesium sulfate shows visible particulates within 14 days. The addition ratio for this format is 0.02–0.08 wt%, with the GHK pre-dissolved as a 1.0 wt% stock in 10% propanediol/water and filtered through 0.22 µm PVDF before final blending. Production under nitrogen blanketing is standard, with residual dissolved oxygen maintained below 1.0 mg/L, and non-preserved ampoules are terminal-filtered through 0.22 µm membrane at 18–25 °C before aseptic filling. Terminal finished product forms include single-dose ampoules, dropper serums, and sheet mask essences. Microbial quality is controlled to ISO 17516:2014 limits, with a total aerobic mesophilic count not exceeding 100 CFU/g for leave-on facial products, and preservative efficacy validated by ISO 11930:2019. The operational boundary is explicit: free copper salts such as copper sulfate are not combined with GHK at pH values above 5.5 because the resulting copper-GHK complex develops a blue tint and can precipitate in low-ionic-strength serums. If a copper-coordinated peptide is required, preformed GHK-Cu should be selected and confirmed by HPLC at 210–220 nm before scale-up.
Filling-line audits reveal that transparent serum clarity failures frequently originate not from the peptide itself but from incompatible rubber plunger lubricants in pre-sterilized syringes, where silicone-oil droplets nucleate peptide aggregation at the droplet-water interface. For this reason, barrier-coated plungers and glass ampoules with low silicone transfer are specified for contract manufacturing batches exceeding 10,000 units. The process window around pH 6.2 is narrow: at pH 6.5, accelerated studies show a measurable reduction in HPLC purity after 4 weeks at 40 °C, while at pH 4.5 the imidazole ring becomes protonated and clarity remains stable but sensory assessed stickiness increases in glycerin-heavy bases above 15 wt%.
A contract manufacturer handling post-procedure aftercare products must first determine whether the finished good is notified as a cosmetic under Regulation (EC) No 1223/2009 or placed on the market as a medical device under Regulation (EU) 2017/745. This single classification changes the addition ratio, cleaning validation, sterility expectation, and terminal test panel. In cosmetic-positioned post-laser aftercare serums, GHK is added at 0.05–0.10 wt% after the bulk has cooled to 18–25 °C, and the process includes 0.22 µm cold filtration, peristaltic pump filling at 50–200 mL/min, and stainless-steel transfer lines passivated to reduce peptide adsorption. Gamma irradiation at 25 kGy is not recommended for aqueous GHK-containing aftercare products because peptide backbone degradation under ionizing radiation is not fully characterized; terminal moist-heat sterilization at 121 °C is also avoided because aqueous GHK degrades measurably above 60 °C during extended hold times. Terminal formats for this use context include hydrogel masks, anhydrous silicone gels, and post-procedure aftercare serums. Cosmetic-grade production is anchored to ISO 22716, ISO 17516:2014, and ISO 11930:2019; medical-device positioning instead invokes ISO 10993-5:2009 for cytotoxicity and ISO 10993-10:2021 for irritation and skin sensitization, with cleanroom filling under ISO 14644-1 class 7 or better. The limitation is regulatory rather than chemical: cosmetic aftercare products cannot claim wound healing, tissue repair, or post-procedure recovery if the stratum corneum is breached; such claims move the finished good into a medical device or drug classification in key export markets.
Published data for this specific post-procedure configuration is limited, and batch release should therefore rely on conservative in-house stability protocols rather than assumed clinical performance. When the aftercare format is an anhydrous silicone gel, the tripeptide is first dissolved in a small amount of propanediol and then dispersed under vacuum; direct powder addition into cyclopentasiloxane leads to agglomerates visible as white specks after 24 h of storage. pH is controlled only in the aqueous phase; anhydrous systems without a buffered water phase require a water activity below 0.60 and hermetic packaging to prevent moisture-driven phase separation.
Rinse-off scalp systems impose a contact time of 2–5 min under normal use, so a higher addition of 0.05–0.20 wt% is necessary to maintain a technically relevant deposited amount after dilution and rinse. In shampoo production, GHK is not added during neutralization because the sodium chloride content and pH rebound across 5.5–7.0 can destabilize the lamellar surfactant phase; the peptide stock is instead added at ≤45 °C after final viscosity adjustment. Cationic polymers such as polyquaternium-10 at 0.3–0.8 wt% form coacervates that bind GHK; in clear scalp tonics, this binding lowers the free peptide concentration in the bulk phase and can require a co-solvent system of 5–10 wt% ethanol and 0.5–1.0 wt% propanediol to maintain clarity at 5 NTU or below. Terminal finished goods in this category include scalp tonics, hair masks, conditioners, and pre-shampoo treatments. Compliance follows Regulation (EC) No 1223/2009 for cosmetic positioning and ISO 22716 for manufacturing; microbial limits reference ISO 17516:2014. The boundary condition is strict: cosmetic scalp products containing GHK cannot claim androgenetic alopecia treatment, hair regrowth, or dermal papilla stimulation in most regulated markets; such language triggers drug or medical device review.
Production-scale records show that clear scalp tonic formulations above 0.10 wt% GHK can develop a faint opalescence when stored at 5 °C for 72 h, especially in formulations containing zinc PCA or magnesium sulfate at levels above 0.2 wt%. This phenomenon is reversible upon warming to 25 °C, but it creates visual rejection complaints if cold-chain shipping is not controlled. Preservative selection also changes peptide recovery: phenoxyethanol at 0.5–0.8 wt% is generally compatible, whereas high levels of certain organic acids below their pKa can protonate the histidine residue and reduce HPLC-detectable GHK in long-term stability samples.
When GHK is loaded into preclinical dermal matrices with water activity above 0.85, peptide stability becomes the primary process variable because unbound water accelerates histidine side-chain oxidation and reduces recovery from the polymer network. Loading at 0.01–0.10% w/w relative to dry polymer mass is achieved by dissolving the tripeptide in the aqueous phase of a collagen, chitosan, or gelatin dope before lyophilization or electrospinning. In electrospinning, high-voltage fields of 15–25 kV and a flow rate of 0.5–1.5 mL/h can expose the peptide to shear and charge-induced orientation; process recovery must be verified by HPLC at 214 nm before and after spinning to detect peptide loss. Terminal formats are research hydrogel dressings, nanofibrous scaffolds, and freeze-dried sponges intended for in vitro or preclinical dermal evaluation. Compliance for non-clinical material follows ISO 10993-5:2009 for cytotoxicity and ISO 10993-12:2021 for sample preparation where the sponsor intends eventual medical device testing; materials supplied only for in vitro studies are typically released under a certificate of analysis within an ISO 9001:2015 quality management system. Published data for specific scaffold configurations is limited, and no universal sterilization compatibility should be assumed for ethylene oxide, gamma irradiation, or electron beam treatment of GHK-loaded dermal matrices.
The operational boundary in this segment is the absence of clinical claims for raw-material lots sold into research use. A dermal matrix containing GHK that is intended for human wound contact must meet the full medical device evaluation program under Regulation (EU) 2017/745, including biological evaluation planning according to ISO 10993-1:2018. In lyophilized matrices, residual moisture above 3.0% after freeze-drying is a common cause of peptide degradation during storage; Karl Fischer titration and differential scanning calorimetry on the cake are therefore used as in-process controls rather than visual inspection alone.
Waterless delivery formats with total water activity below 0.60 reject direct powder addition of GHK because undissolved particles survive filling and create visible particulates in clear formulas. Where a waterless serum or balm is manufactured, the tripeptide is pre-dispersed as a 1.0 wt% solution in 70:30 propanediol/butylene glycol, then incorporated at 0.01–0.05 wt% final concentration under vacuum at 25–35 °C with 50–100 rpm anchor mixing. The production environment requires humidity control below 60% RH; high-humidity exposure above this threshold introduces water into the glycol stock and can shift the water activity upward, compromising the anhydrous positioning. Filling is conducted under nitrogen, and airless packaging with low headspace is used to limit oxygen ingress. Terminal finished product types include waterless serums, touch pens, anhydrous balms, and powder-to-emulsion kits where GHK is kept in a separate low-water phase until the consumer reconstitutes the product. Compliance for these anhydrous cosmetics remains under Regulation (EC) No 1223/2009 and ISO 22716; preservation strategy must be re-evaluated if the final water activity rises above 0.60, at which point ISO 11930:2019 challenge testing becomes relevant even if the base formula is nominally water-free.
Process audits identify the glycol pre-dispersion step as the primary batch bottleneck because the tripeptide dissolves slowly in propanediol-rich media below 30 °C; preheating the glycol blend to 40–45 °C before peptide addition reduces dissolution time but must be followed by cooling to 25–35 °C before compounding with heat-sensitive oils. In waterless bases containing more than 50 wt% volatile silicones, GHK stock separation can occur at the filling valve if the recirculation loop is not maintained above 50 rpm, producing inconsistent peptide content across fill heads. Limitations in this segment are not regulatory but physical: the tripeptide remains a water-loving molecule, and its stability in anhydrous systems depends entirely on preventing localized moisture pockets during compounding and storage.
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Glycyl-L-Histidyl-L-Lysine (GHK; CAS 49557-75-7) is a synthetic tripeptide of sequence Gly-His-Lys with molecular formula C14H24N6O4 and monoisotopic mass 340.1859 Da. The product is usually supplied as a lyophilized acetate salt, although hydrochloride and free-base presentations are encountered. Supplier catalog model designations typically encode salt form, net peptide content, fill mass, and packaging format; these designations are not structural identifiers and must be cross-referenced with the lot-specific certificate of analysis. Because the acetate counterion and residual water are not part of the net peptide mass, formulators must convert total powder mass to net peptide mass using amino acid analysis or equivalent HPLC-derived data before preparing high-precision solutions. The material is a white to off-white powder, hygroscopic after opening, and should be equilibrated to ambient temperature before exposure to avoid condensation-driven hydration. Standard storage is −20 °C ± 5 °C in a desiccated atmosphere; lyophilized powder should be protected from repeated temperature excursions that can produce amorphous-to-crystalline shifts and change reconstitution behavior.
Release criteria for GHK are based on orthogonal methods because lyophilized peptide salts contain counterions and residual water that are not part of the peptide mass. A typical release specification uses reversed-phase HPLC per USP 621 for purity, electrospray ionization mass spectrometry for identity, and Karl Fischer titration per USP 921 for water content. Cell-culture and tissue-engineering grades additionally control endotoxin by the limulus amebocyte lysate method per USP 85. Because acetate is not detected by the HPLC purity method at 214 nm, ion chromatography is used to report the counterion content. The following matrix summarizes typical release attributes.
| Attribute | Method | Typical release limit |
|---|---|---|
| Appearance | Visual inspection | White to off-white lyophilized powder |
| Identity | ESI-MS | [M+H]+ at m/z 341.19 |
| Purity | RP-HPLC (USP 621) | ≥98.0% area at 214 nm |
| Net peptide content | Amino acid analysis or HPLC | 75–90% for acetate salt; lot-dependent |
| Acetate content | Ion chromatography | Report result |
| Water content | Karl Fischer (USP 921) | ≤5.0% |
| Endotoxin | LAL (USP 85) | ≤0.5 EU/mg for research/cell grade |
| Residual solvents | GC (USP 467) | Report result |
Because peptide purity and net peptide content are not identical, a powder with 98.0% HPLC purity may contain only 80% peptide by mass once acetate and water are accounted for. Formulators should calculate GHK mass from net peptide content rather than total powder weight. The acetate salt has a different mass balance than the hydrochloride; interchanging salt forms without recalculation changes final molarity and counterion concentration. A typical HPLC release method uses a C18 column with a water/acetonitrile gradient containing 0.1% trifluoroacetic acid and detection at 214 nm. Published data for specific column dimensions and gradient slopes vary by supplier and should be obtained from the certificate of analysis.
GHK is freely soluble in water and phosphate-buffered saline. Aqueous stock solutions at 1–10 mg/mL are prepared with low-shear mixing; the powder should be added to the vortex of water for injection or buffer to avoid floating partially hydrated aggregates. High-shear homogenization exceeding 3,000 rpm can increase air entrainment and peptide adsorption at gas-liquid interfaces, reducing recovery; published data for this specific configuration is limited. pH adjustment with dilute sodium hydroxide or hydrochloric acid should keep the solution between pH 5.5 and 7.0. Lyophilization from water or 5 mM ammonium acetate is preferred for long-term storage; phosphate buffers are less suitable for freezing because eutectic crystallization can induce local pH shifts. Solutions for cell-culture use are sterilized by filtration through 0.22 µm polyethersulfone membranes. Autoclaving is not recommended because thermal hydrolysis of the His-Lys peptide bond may occur; published data for this specific configuration is limited.
In cold-process aqueous serum and gel systems, GHK is introduced as a pre-dissolved stock at the final formulation stage. Copper-free systems are preferred because the free base can sequester Cu(II) from equipment surfaces and raw materials, forming a blue GHK-Cu complex that may be visually detectable against a white emulsion background. This chelation behavior means that GHK should not be mixed with copper-containing traces in the same vessel without verifying final color. In vitro fibroblast and keratinocyte models often use GHK concentrations of 0.1 nM to 1 µM; cosmetic dosage is product-specific and requires stability and safety substantiation under Regulation (EC) 1223/2009. Preservation efficacy should be verified by ISO 11930 because peptide-containing formulations can alter preservative partitioning and reduce challenge-test pass rates if the peptide fraction binds preservative molecules. The free tripeptide is not a pharmacologically classified drug substance and should not be assigned drug claims in cosmetic or research documentation.
GHK is the uncomplexed tripeptide and should not be confused with Copper Tripeptide-1 (GHK-Cu). The copper complex carries pre-coordinated Cu(II), has a blue to purplish-blue solid appearance, and is selected when copper-dependent in vitro responses are the object of study. Palmitoyl Tripeptide-1 is the N-palmitoylated derivative of GHK; the palmitoyl chain increases oil solubility and skin-partitioning behavior. The free base is smaller, more water-soluble, and does not introduce the redox or copper-related variables associated with the copper complex. The following comparison distinguishes the three materials as raw-material classes.
| Parameter | GHK free base | GHK-Cu | Palmitoyl Tripeptide-1 |
|---|---|---|---|
| CAS | 49557-75-7 | 89030-95-5 | — |
| Solid appearance | White to off-white powder | Blue to purplish-blue solid | White to off-white waxy solid |
| Molecular mass | 340.19 Da | Approx. 401.9 Da | Approx. 578.8 Da |
| Metal coordination | None as supplied | Pre-coordinated Cu(II) | None as supplied |
| Water solubility | High | Moderate | Low |
| Preferred formulation phase | Aqueous | Aqueous | Oil or emulsion oil phase |
The choice between GHK and its derivatives depends on formulation phase and target. The free base is preferable when copper ion delivery is not intended and when compatibility with chelating agents is required. GHK-Cu is preferred where copper-dependent in vitro responses are the focus. The palmitoylated derivative is used where increased lipophilicity and resistance to wash-off are required. All three forms should be considered distinct raw materials with separate stability data, hazardous substance registration, and formulation limits; substitution without reformulation is not acceptable.
GHK is incompatible with strong oxidizers, amine-reactive crosslinkers, and prolonged exposure to elevated pH. Above pH 8.0, histidine deprotonation can alter copper-binding affinity and increase hydrolysis; below pH 2.0, acid-catalyzed hydrolysis of peptide bonds is the main risk. Avoid combination with carbodiimide coupling agents unless the peptide is intended as a reactant. In emulsion systems, GHK partitions into the aqueous phase; if added to the hot oil phase above 40 °C, recovery loss may occur. Processing equipment should be passivated 316L stainless steel or glass-lined, because copper ions leached from unpassivated copper-containing alloys can form GHK-Cu and change color. After batch mixing, rinse water should be assayed by HPLC to quantify adsorbed peptide; if recovery is below 90%, static adsorption is suspected and a pre-rinse with dilute GHK solution or a pH 5.5 citrate buffer may be used. Batch-to-batch variance in net peptide content is managed by net peptide assay rather than total powder weight, and any formulation record should record net peptide mass, salt form, and water content.
