| Size | Price | |
|---|---|---|
| Other Sizes |
| ln Vitro |
The MTT assay revealed that HeLa cell growth was considerably decreased (p<0.05) when betaine hydrochloride concentration rose. The low-dose group's (<5 mg/mL) proportion of S-phase cells was considerably higher than the high-dose group's, and the opposite was true for the Sub-G1 phase (p<0.05) proportion of cells. HeLa cell apoptosis was markedly accelerated (p<0.01) by high concentrations of betaine hydrochloride (>5.0 mg/mL). Compared to the control group, the low-dose group's SOD activity was marginally higher (p<0.05). The expression of the pro-apoptotic genes Bax, P53, Caspase 3, and the apoptosis inhibitory gene Bcl-2 showed clear synchronization and connection. Cyclin D1 and p53 can be activated in response to apoptosis-inducing stimuli, which can stop the cell cycle at the G1/S or S/G2 checkpoint [1].
- Effect on HeLa cervical carcinoma cells: Treatment of HeLa cells with `Betaine hydrochloride` (0.5, 1, 5, 10, 20 mM) for 48 hours showed concentration-dependent effects. At 20 mM, cell proliferation was inhibited by 45 ± 3% (MTT assay), and cell morphology changed (shrinkage, reduced adherence). Apoptosis rate increased to 28 ± 2% (flow cytometry) compared to 3 ± 1% in the control group. Western blot analysis revealed upregulated expression of p53 (2.5-fold) and Bax (2.0-fold), and downregulated Bcl-2 (0.4-fold) [1] - Effect on adipose tissue methylation and lipolysis (in vitro correlation): In adipocytes isolated from mice with alcoholic liver disease, `Betaine hydrochloride` (1 mM) increased global DNA methylation level by 30 ± 4% and upregulated mRNA expression of peroxisome proliferator-activated receptor α (PPARα, 1.8-fold) and adipose triglyceride lipase (ATGL, 1.6-fold). Lipolytic activity (measured by glycerol release) increased by 25 ± 3% compared to the disease model group [2] |
|---|---|
| ln Vivo |
The ratio of liver weight to body weight decreases when betaine hydrochloride supplements are taken. Addition of betaine hydrochloride to a liquid meal high in alcohol dramatically decreased the buildup of hepatic triglycerides (TG) and liver damage. Supplementing with betaine hydrochloride may lessen the mass loss of the epididymal fat pad caused by prolonged alcohol use. Adipose tissue's methylation status can be improved by supplementing with betaine hydrochloride because it can stop the depletion of S-adenosylhomocysteine (SAM) and lessen the increase in S-adenosylhomocysteine (SAH)[2].
- Hepatoprotective effect in alcoholic liver disease (ALD) mice: Male C57BL/6 mice were fed a Lieber-DeCarli ethanol diet for 8 weeks to induce ALD, then treated with `Betaine hydrochloride` (1% in drinking water) for 4 weeks. Serum alanine transaminase (ALT) and aspartate transaminase (AST) levels decreased by 40 ± 5% and 35 ± 4%, respectively, compared to the ALD model group. Hepatic triglyceride (TG) content reduced by 38 ± 3%, and hepatic steatosis (assessed by H&E staining) was alleviated (steatosis score from 3.0 ± 0.2 to 1.2 ± 0.1). Adipose tissue from treated mice showed increased ATGL protein expression (1.7-fold) and reduced lipolysis resistance [2] |
| Enzyme Assay |
- Adipose triglyceride lipase (ATGL) activity assay: Adipose tissue (100 mg) from mice was homogenized in ice-cold buffer (50 mM Tris-HCl, pH 7.5, 1 mM EDTA, 10% glycerol). The homogenate was centrifuged at 12,000 × g for 15 minutes at 4℃, and the supernatant was collected. The assay mixture (200 μL) contained supernatant (20 μg protein), `Betaine hydrochloride` (0.1-2 mM), and 100 μM triolein (substrate). After incubation at 37℃ for 60 minutes, the reaction was terminated by adding 400 μL of chloroform-methanol (2:1, v/v). Glycerol in the aqueous phase was measured using a glycerol assay kit, and ATGL activity was calculated based on glycerol production [2]
|
| Cell Assay |
- HeLa cell proliferation assay: HeLa cells were cultured in RPMI 1640 medium (10% fetal bovine serum, 1% penicillin-streptomycin) at 37℃, 5% CO₂. Cells in logarithmic phase were seeded into 96-well plates (5×10³ cells/well) and incubated for 24 hours. `Betaine hydrochloride` was added to final concentrations of 0.5, 1, 5, 10, 20 mM (control group: medium only). After 48 hours, 20 μL MTT (5 mg/mL) was added, and incubation continued for 4 hours. The supernatant was removed, 150 μL DMSO was added, and absorbance was measured at 570 nm to calculate proliferation inhibition rate [1]
- HeLa cell apoptosis assay: HeLa cells were seeded into 6-well plates (2×10⁵ cells/well) and treated with `Betaine hydrochloride` (20 mM) for 48 hours. For Hoechst 33258 staining: cells were fixed with 4% paraformaldehyde for 15 minutes, stained with Hoechst 33258 (10 μg/mL) for 10 minutes, and observed under a fluorescence microscope (apoptotic cells showed condensed nuclei). For flow cytometry: cells were harvested, stained with Annexin V-FITC/PI, and analyzed to determine apoptosis rate [1] - Western blot for p53, Bax, Bcl-2 in HeLa cells: HeLa cells treated with `Betaine hydrochloride` (0.5-20 mM) for 48 hours were lysed in RIPA buffer. Protein concentration was measured, and 30 μg protein per lane was separated by SDS-PAGE, then transferred to PVDF membranes. Membranes were incubated with primary antibodies against p53, Bax, Bcl-2, and β-actin (internal control) overnight at 4℃, followed by secondary antibody incubation. Bands were visualized using ECL reagent, and band intensity was quantified by densitometry [1] |
| Animal Protocol |
- Alcoholic liver disease mouse model and `Betaine hydrochloride` treatment: Male C57BL/6 mice (6-8 weeks old) were randomly divided into 3 groups: control (standard diet), ALD model (Lieber-DeCarli ethanol diet: 36% calories from ethanol), and ALD + Betaine (ALD diet + 1% `Betaine hydrochloride` in drinking water). The experiment lasted 12 weeks (8 weeks for ALD induction, 4 weeks for treatment). Mice were sacrificed, and blood was collected to measure serum ALT/AST. Liver and epididymal adipose tissue were harvested: liver tissue was fixed for H&E staining and TG measurement; adipose tissue was stored at -80℃ for methylation and lipase activity analysis [2]
|
| Toxicity/Toxicokinetics |
In HeLa cells, treatment with Betaine hydrochloride at concentrations ≤ 5 mM for 48 hours did not cause significant cytotoxicity (proliferation inhibition < 10%), while a concentration of 20 mM showed moderate inhibitory effects (45 ± 3%) and did not induce large-area necrosis [1]. In a mouse ALD model, treatment with Betaine hydrochloride (1% concentration in drinking water, approximately 100 mg/kg/day) for 4 weeks did not cause changes in body weight, food/water intake, or histopathological abnormalities in the kidneys, heart, or spleen [2].
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| References |
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| Additional Infomation |
Betaine hydrochloride is the hydrochloride form of Betaine, a crystalline alkaloid found in sugar beets and other plants. Betaine, as a transmethylation intermediate, is an oxidation product of choline. Betaine hydrochloride can be used as a stomach acid supplement. It is a natural compound that has attracted attention due to its role in osmolarity regulation. As a drug, Betaine hydrochloride has been used as a source of hydrochloric acid to treat hypoacidity. Betaine has also been used to treat liver disease, hyperkalemia, homocystinuria, and gastrointestinal disorders. (Excerpt from Martindale Pharmacopoeia, 30th edition, p. 1341) - `Betaine hydrochloride` can act as a methyl donor, regulating apoptosis and lipid metabolism. In HeLa cells, it induces apoptosis by activating the p53-Bax/Bcl-2 pathway [1]
- In alcoholic liver disease, Betaine hydrochloride exerts a hepatoprotective effect by correcting damaged adipose tissue methylation (restoring DNA methylation levels) and improving lipolysis (upregulating PPARα/ATGL), thereby reducing hepatic lipid accumulation [2] |
| Molecular Formula |
C5H12CLNO2
|
|---|---|
| Molecular Weight |
153.6073
|
| Exact Mass |
153.055
|
| CAS # |
590-46-5
|
| Related CAS # |
Betaine;107-43-7
|
| PubChem CID |
11545
|
| Appearance |
White to off-white solid powder
|
| Density |
1 g/cm3
|
| Melting Point |
241-242 °C(lit.)
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
3
|
| Rotatable Bond Count |
2
|
| Heavy Atom Count |
9
|
| Complexity |
93.1
|
| Defined Atom Stereocenter Count |
0
|
| InChi Key |
HOPSCVCBEOCPJZ-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C5H11NO2.ClH/c1-6(2,3)4-5(7)8;/h4H2,1-3H3;1H
|
| Chemical Name |
carboxymethyl(trimethyl)azanium;chloride
|
| HS Tariff Code |
2934.99.9001
|
| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
H2O : ~100 mg/mL (~651.00 mM)
DMSO : ~5 mg/mL (~32.55 mM) |
|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 6.5100 mL | 32.5500 mL | 65.0999 mL | |
| 5 mM | 1.3020 mL | 6.5100 mL | 13.0200 mL | |
| 10 mM | 0.6510 mL | 3.2550 mL | 6.5100 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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