Cat. No.: HY-W013699

Chlorhexidine diacetate

13 Cited Publications

Customer Review

Based on 13 publication(s) in Google Scholar

Chlorhexidine diacetate is a orally active cationic antimicrobial agent that targets microbial cell membranes. Chlorhexidine diacetate binds to cell membrane phospholipids non-specifically, destroys membrane structure and induces leakage of cell contents. Chlorhexidine diacetate has broad-spectrum bactericidal activity against both Gram-positive and Gram-negative bacteria. Chlorhexidine diacetate can interfere with membrane permeability, cause protein precipitation and energy metabolism disorders, such as rapid inhibition of microbial growth and induction of cell death (necrosis or apoptosis).

For research use only. We do not sell to patients.

Purity: 99.68%
CAS No.: 56-95-1
Formula: C₂₆H₃₈Cl₂N₁₀O₄
Molecular Weight:625.55

Storage:

4°C, stored under nitrogen

* In solvent : -80°C, 6 months; -20°C, 1 month (stored under nitrogen)

Chlorhexidine diacetate Chemical Structure

CAS No.: 56-95-1

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Biological Activity
- Description
- In Vitro
- In Vivo
Chemical Information
Publications (13)
Solvent & Solubility
- In Vitro
- In Vivo
Purity & Documentation
References
Help & FAQs
Classifications
Bioz

Biological Activity

Description

All Antibiotic Isoforms :

In Vitro

1. Cell death assay:
Chlorhexidine diacetate (0.000125%-0.016%; 24 h) induces cell death in L929 fibroblasts, inducing a pattern of cell necrosis and/or apoptosis, as well as cell stress. Furthermore, Chlorhexidine diacetate induces apoptosis at lower concentrations and necrosis at higher concentrations, and increases the expression of heat shock protein 70 (a marker of cell stress). Chlorhexidine diacetate may have an adverse effect on the resolution of apical periodontitis^[1].
2. Cell viability experiment:
Chlorhexidine diacetate (0.0005%-0.5%; 30 min) causes the death of canine embryonic fibroblasts with concentration of ≥0.013%, while allows the cells to survive with concentration of ≤0.006%^[2].
3. Bacterial survival experiment:
Chlorhexidine diacetate (0.0005%-0.5%; 30 min) has a concentration-dependent bactericidal effect on Staphylococcus aureus, and kills Staphylococcus aureus with concentration of ≥0.05%, while allows the bacteria to survive with concentration of ≤0.03%^[2].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Cell Viability Assay^[3]

Cell Line:

Canine embryonic fibroblasts
Concentration:

0.5%, 0.05%, 0.03%, 0.013%, 0.006%, 0.005%, 0.0005%
Incubation Time:

30 minutes
Result:

Fibroblasts showed 0% survival at concentrations ≥0.013%, while concentrations ≤0.006% allowed significant survival (e.g., 74% survival at 0.0005%), indicating a concentration-dependent cytotoxic effect.

In Vivo

1. Paw injection toxicity test:
Chlorhexidine (0.125%-1.0%; subcutaneous injection; single dose) diacetate causes toxic effects in the plantar space of the hind paw of Balb/c mice. Concentrations ≥0.5% induces coagulative necrosis of the epidermis, dermis and subcutaneous tissues and neutrophil-dominated inflammatory responses, while concentrations ≤0.25% induces moderate inflammation and interstitial edema^[1].
2. Lung inhalation toxicity test:
Chlorhexidine (0.125%-1%; intratracheal instillation; single dose; 8-week observation) diacetate causes collagen fiber deposition and inflammatory cell infiltration in lung tissue in the C57BL/6J mouse model, leading to restrictive ventilation dysfunction. Transcriptome analysis shows that genes related to extracellular matrix deposition and mucus secretion (such as Muc5b and Muc5ac) are upregulated^[3].
3. Genetic toxicity test:
0.12% Chlorhexidine dihydrochloride (0.5 mL; oral, twice a day for 8 days) causes DNA damage in peripheral blood and oral mucosal cells of rats, without causing chromosome breakage or loss in erythrocytes^[4].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Animal Model:

Male Balb/c mice (4 weeks old, 20 g), paw subplantar injection model^[1]
Dosage:

0.125%, 0.25%, 0.5%, 1.0% chlorhexidine in distilled water
Administration:

Subplantar injection into the right hind paw, single dose, observed at 24 h, 48 h, 7 days, and 14 days
Result:

At 0.5% and 1.0%, caused extensive coagulative necrosis in the epidermis, dermis, and subcutaneous tissue, associated with marked neutrophilic inflammation and edema at 24-48 h; 0.25% induced focal necrosis in few specimens and moderate inflammation in all; 0.125% caused no necrosis but moderate neutrophilic inflammation and edema.
Epidermal healing was complete by 7-14 days, with residual mononuclear cell infiltration and fibrous tissue thickening in high-dose groups.

Animal Model:

C57BL/6J mice (6-8 weeks old), intratracheal instillation-induced lung injury model^[2]
Dosage:

0.125% (2.5 mg/kg), 0.25% (5 mg/kg), 0.5% (10 mg/kg), 1% (20 mg/kg) chlorhexidine gluconate in RO water
Administration:

Single intratracheal instillation via MicroSprayer Aerosolizer, observed for 8 weeks
Result:

Led to dose-dependent lung tissue damage, including inflammatory cell infiltration, alveolar protein deposition, and airway epithelial cell exfoliation.
Increased collagen fiber deposition in the alveolar septum and bronchial stroma, with significantly higher lung organ coefficients in high-dose groups.
Decreased respiratory system compliance (Crs) and forced vital capacity (FVC), indicating restrictive ventilatory dysfunction.
Upregulated genes related to extracellular matrix organization (e.g., P4ha3) and mucus secretion (e.g., Muc5b, Muc5ac).

Chemical Information

CAS No. 56-95-1
Appearance Solid
Molecular Weight 625.55
Formula C₂₆H₃₈Cl₂N₁₀O₄
Color White to off-white
SMILES

N=C(NC1=CC=C(Cl)C=C1)NC(NCCCCCCNC(NC(NC2=CC=C(Cl)C=C2)=N)=N)=N.CC(O)=O.CC(O)=O
Shipping

Room temperature in continental US; may vary elsewhere.
Storage

4°C, stored under nitrogen

* In solvent : -80°C, 6 months; -20°C, 1 month (stored under nitrogen)

Publications (13)

All WB

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Journal Impact Factor

Nat Commun

The folate cycle enzyme MTHFD2 induces cancer immune evasion through PD-L1 up-regulation. [Abstract]

2021 Mar 29;12(1):1940. PMID: 33782411
Cell Death Dis

HJURP regulates cell proliferation and chemo-resistance via YAP1/NDRG1 transcriptional axis in triple-negative breast cancer. [Abstract]

2022 Apr 22;13(4):396. PMID: 35459269
Antioxidants (Basel)

Pitavastatin Calcium Confers Fungicidal Properties to Fluconazole by Inhibiting Ubiquinone Biosynthesis and Generating Reactive Oxygen Species. [Abstract]

2024 May 29;13(6):667. PMID: 38929106
Dent Mater

Triple-modified PEEK surface via plasma treatment, polydopamine coating and chlorhexidine: Assessment of biocompatibility and antibacterial properties. [Abstract]

2025 Apr 17:S0109-5641(25)00607-4. PMID: 40251086
Cell Oncol (Dordr)

UBE2Q2 promotes tumor progression and glycolysis of hepatocellular carcinoma through NF-κB/HIF1α signal pathway. [Abstract]

2025 Jun;48(3):637-654. PMID: 39833608
Mol Oncol

Loss of RDM1 enhances hepatocellular carcinoma progression via p53 and Ras/Raf/ERK pathways. [Abstract]

2020 Feb;14(2):373-386. PMID: 31670863
Front Cell Dev Biol

SYVN1-MTR4-MAT2A Signaling Axis Regulates Methionine Metabolism in Glioma Cells. [Abstract]

2021 Mar 30:9:633259. PMID: 33859984
Transl Oncol

20-hydroxyecdysone suppresses bladder cancer progression via inhibiting USP21: A mechanism associated with deubiquitination and degradation of p65. [Abstract]

2024 Jul:45:101958. PMID: 38663220
J Funct Foods

2024 Jun.
Cell Signal

pVHL promotes lysosomal degradation of YAP in lung adenocarcinoma. [Abstract]

2021 Jul:83:110002. PMID: 33823241
Genes (Basel)

Autosomal Recessive Retinitis Pigmentosa Associated with Three Novel REEP6 Variants in Chinese Population. [Abstract]

2021 Apr 7;12(4):537. PMID: 33917198
Biochem Biophys Res Commun

Mechanisms of NMDA receptor inhibition by chlorhexidine and alexidine. [Abstract]

2025 Dec 17:796:153158. PMID: 41418346
bioRxiv

2024 Nov 06.

Solvent & Solubility

In Vitro:

DMSO : ≥ 100 mg/mL (159.86 mM; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Ethanol : 55 mg/mL (87.92 mM; Need ultrasonic)

H₂O : 20 mg/mL (31.97 mM; ultrasonic and warming and heat to 60°C)

* "≥" means soluble, but saturation unknown.

View the Complete Stock Solution Preparation Table

Preparing Stock Solutions

View the Complete Stock Solution Preparation Table

Concentration Solvent Mass

1 mg

5 mg

10 mg

Concentration

Solvent

Mass

1 mM

1.5986 mL

7.9930 mL

15.9859 mL

5 mM

0.3197 mL

1.5986 mL

3.1972 mL

10 mM

0.1599 mL

0.7993 mL

1.5986 mL

Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.

Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (stored under nitrogen). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

Molarity Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

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Molecular Weight Molecular Weight

Dilution Calculator

Concentration _(start) × Volume _(start) = Concentration _(final) × Volume _(final)

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In Vivo:

Select the appropriate dissolution method based on your experimental animal and administration route.

Stock Solution Working Solution

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for In Vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

Protocol 1

Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline

Solubility: ≥ 2.5 mg/mL (4.00 mM); Clear solution

This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

Protocol 2

Add each solvent one by one: 10% DMSO 90% (20% SBE-β-CD in Saline)

Solubility: ≥ 2.5 mg/mL (4.00 mM); Clear solution

This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.

Protocol 3

Add each solvent one by one: 10% DMSO 90% Corn Oil

Solubility: 2.5 mg/mL (4.00 mM); Suspended solution; Need ultrasonic

This protocol yields a suspended solution of 2.5 mg/mL. Suspended solution can be used for oral and intraperitoneal injection.

Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL Corn oil, and mix evenly.

Protocol 4

Add each solvent one by one: 10% EtOH 40% PEG300 5% Tween-80 45% Saline

Solubility: ≥ 2.5 mg/mL (4.00 mM); Clear solution

This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

Taking 1 mL working solution as an example, add 100 μL EtOH stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

Protocol 5

Add each solvent one by one: 10% EtOH 90% (20% SBE-β-CD in Saline)

Solubility: ≥ 2.5 mg/mL (4.00 mM); Clear solution

This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

Taking 1 mL working solution as an example, add 100 μL EtOH stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.

Protocol 6

Add each solvent one by one: 10% EtOH 90% Corn Oil

Solubility: ≥ 2.5 mg/mL (4.00 mM); Clear solution

This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown). If the continuous dosing period exceeds half a month, please choose this protocol carefully.

Taking 1 mL working solution as an example, add 100 μL EtOH stock solution (25.0 mg/mL) to 900 μL Corn oil, and mix evenly.

In Vivo Dissolution Calculator

Please enter the basic information of animal experiments:

Dosage Dosage
mg/kg
Animal Weight (Per Animal) Animal Weight (Per Animal)
g
Dosing Volume (Per Animal) Dosing Volume (Per Animal)
μL
Number of Animals Number of Animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.

Working solution concentration: 0.22 mg/mL

This product has good water solubility, please refer to the measured solubility data in water/PBS/Saline for details.

The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only.If necessary, please contact

MedChemExpress (MCE) .

Purity & Documentation

Select Batch:

Purity: 99.98%

Assay: 99.98%

ee.: 99.98%

Data Sheet (278 KB)
SDS (623 KB)
- English - EN (623 KB)
- Français - FR (623 KB)
- Deutsch - DE (623 KB)
- Norwegian - NO (623 KB)
- Español - ES (623 KB)
- Swedish - SV (623 KB)
- Italian - IT (623 KB)
- Korean - KR (623 KB)
- Portuguese - PT (623 KB)
COA (2659 KB)
Elemental Analysis Report (2659 KB)
HNMR (2659 KB)
CNMR (2659 KB)
RP-HPLC (2659 KB)
NP-HPLC (2659 KB)
MS (2659 KB)
LCMS (2659 KB)
IR (2659 KB)
GC (2659 KB)
OR (2659 KB)
SDS-PAGE (2659 KB)
SEC-HPLC (2659 KB)
Other Report (2659 KB)
Handling Instructions (2659 KB)

References

[1]. Faria G, et al. Evaluation of chlorhexidine toxicity injected in the paw of mice and added to cultured l929 fibroblasts. J Endod. 2007 Jun;33(6):715-22. [Content Brief]

[2]. Sanchez IR, et al. Chlorhexidine diacetate and povidone-iodine cytotoxicity to canine embryonic fibroblasts and Staphylococcus aureus. Vet Surg. 1988;17(4):182-185. [Content Brief]

[3]. Zhang J, et al. Pulmonary Toxicity Assessment after a Single Intratracheal Inhalation of Chlorhexidine Aerosol in Mice. Toxics. 2023 Nov 7;11(11):910. [Content Brief]

[4]. Ribeiro DA, et al. Chlorhexidine induces DNA damage in rat peripheral leukocytes and oral mucosal cells. J Periodontal Res. 2004 Oct;39(5):358-61. [Content Brief]

Help & FAQs

How should lyophilized recombinant proteins be reconstituted and stored?

1. Before opening the cap, centrifuge the vial at 13000 rpm for 20-30 seconds. This step will ensure that any lyophilized powder that may have adhered to the cap or walls is collected at the bottom of the vial, minimizing the risk of product loss.

Complete Stock Solution Preparation Table

Optional Solvent	Concentration Solvent Mass	1 mg	5 mg	10 mg	25 mg
H₂O / Ethanol / DMSO	1 mM	1.5986 mL	7.9930 mL	15.9859 mL	39.9648 mL
	5 mM	0.3197 mL	1.5986 mL	3.1972 mL	7.9930 mL
	10 mM	0.1599 mL	0.7993 mL	1.5986 mL	3.9965 mL
	15 mM	0.1066 mL	0.5329 mL	1.0657 mL	2.6643 mL
	20 mM	0.0799 mL	0.3996 mL	0.7993 mL	1.9982 mL
	25 mM	0.0639 mL	0.3197 mL	0.6394 mL	1.5986 mL
	30 mM	0.0533 mL	0.2664 mL	0.5329 mL	1.3322 mL
Ethanol / DMSO	40 mM	0.0400 mL	0.1998 mL	0.3996 mL	0.9991 mL
	50 mM	0.0320 mL	0.1599 mL	0.3197 mL	0.7993 mL
	60 mM	0.0266 mL	0.1332 mL	0.2664 mL	0.6661 mL
	80 mM	0.0200 mL	0.0999 mL	0.1998 mL	0.4996 mL
DMSO	100 mM	0.0160 mL	0.0799 mL	0.1599 mL	0.3996 mL

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

Keywords

Chlorhexidine diacetate

13 Cited Publications

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Chlorhexidine diacetate

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Publications Citing Use of MedChemExpress (MCE) Chlorhexidine diacetate

All Antibiotic Isoforms

Biological Activity

Chemical Information

Publications (13)

Solvent & Solubility

Purity & Documentation

References

Help & FAQs

Complete Stock Solution Preparation Table

Chlorhexidine diacetate Related Classifications

HY-W013699 Related Classifications