2. Anti-infection Apoptosis
  3. Antibiotic Bacterial Necroptosis Apoptosis
  4. Chlorhexidine

Chlorhexidine is a orally active cationic antimicrobial agent that targets microbial cell membranes. Chlorhexidine binds to cell membrane phospholipids non-specifically, destroys membrane structure and induces leakage of cell contents. Chlorhexidine has broad-spectrum bactericidal activity against both Gram-positive and Gram-negative bacteria. Chlorhexidine 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).

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Chlorhexidine Chemical Structure

Chlorhexidine Chemical Structure

CAS No. : 55-56-1

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Based on 11 publication(s) in Google Scholar

Other Forms of Chlorhexidine:

Chlorhexidine Related Antibodies

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Description

Chlorhexidine is a orally active cationic antimicrobial agent that targets microbial cell membranes. Chlorhexidine binds to cell membrane phospholipids non-specifically, destroys membrane structure and induces leakage of cell contents. Chlorhexidine has broad-spectrum bactericidal activity against both Gram-positive and Gram-negative bacteria. Chlorhexidine 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)[1][2][3].

In Vitro

1. Cell death assay:
Chlorhexidine (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 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 may have an adverse effect on the resolution of apical periodontitis[1].
2. Cell viability experiment:
Chlorhexidine (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 (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.

Chlorhexidine Related Antibodies

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 min
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) 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) 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:
Chlorhexidine (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).
Clinical Trial
Molecular Weight

505.45

Formula

C22H30Cl2N10
CAS No.
Appearance

Solid

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
Shipping

Room temperature in continental US; may vary elsewhere.
Storage

4°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

Solvent & Solubility
In Vitro: 

DMSO : 25 mg/mL (49.46 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 1.9784 mL 9.8922 mL 19.7843 mL
5 mM 0.3957 mL 1.9784 mL 3.9569 mL
View the Complete Stock Solution Preparation Table

* 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 (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

  • Molarity Calculator

  • Dilution Calculator

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

Mass
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Concentration
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Volume
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Molecular Weight *

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

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start)

C1

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Volume (start)

V1

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C2

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Volume (final)

V2

In Vivo:

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

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 (4.95 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.

    Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
  • Protocol 2

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

    Solubility: ≥ 2.5 mg/mL (4.95 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.

    Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

g

Dosing volume
(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
%
DMSO +
% 
+
%
Tween-80 +
%
Saline
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Calculation results:
Working solution concentration: mg/mL
Method for preparing stock solution: mg drug dissolved in μL  DMSO (Stock solution concentration: mg/mL).

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

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).
Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.
 If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Purity & Documentation
References

Complete Stock Solution Preparation Table

* 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 (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
DMSO 1 mM 1.9784 mL 9.8922 mL 19.7844 mL 49.4609 mL
5 mM 0.3957 mL 1.9784 mL 3.9569 mL 9.8922 mL
10 mM 0.1978 mL 0.9892 mL 1.9784 mL 4.9461 mL
15 mM 0.1319 mL 0.6595 mL 1.3190 mL 3.2974 mL
20 mM 0.0989 mL 0.4946 mL 0.9892 mL 2.4730 mL
25 mM 0.0791 mL 0.3957 mL 0.7914 mL 1.9784 mL
30 mM 0.0659 mL 0.3297 mL 0.6595 mL 1.6487 mL
40 mM 0.0495 mL 0.2473 mL 0.4946 mL 1.2365 mL
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Chlorhexidine Related Classifications

Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

Keywords:

Chlorhexidine55-56-1AntibioticBacterialNecroptosisApoptosisInhibitorinhibitorinhibit

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