| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
Hyodeoxycholic acid targets TGR5 (also known as GPCR19, or G protein-coupled bile acid receptor 1), a cell surface receptor that is activated by various bile acids. In CHO cells, hyodeoxycholic acid acts as a TGR5 agonist with an EC50 of 31.6 microM. Activation of TGR5 leads to increased intracellular cAMP levels, which in turn regulates energy homeostasis, glucose metabolism, and immune function. Hyodeoxycholic acid-d5 retains the same target specificity.
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| ln Vitro |
Drug compounds have included stable heavy isotopes of carbon, hydrogen, and other elements, mostly as tracers for quantification throughout the drug development process. Due to its potential to alter the pharmacokinetic and metabolic characteristics of medications, deuteration has drawn attention[1].
Hyodeoxycholic acid-d5 is not typically used in cell-free enzyme assays because it is an analytical standard rather than a test compound. However, the unlabeled hyodeoxycholic acid can be evaluated for TGR5 binding using cell-free membrane-based binding assays with radiolabeled ligands or TR-FRET technology. The d5-labeled version serves as an internal standard for quantifying HDCA in such assays by LC-MS/MS. |
| ln Vivo |
In cellular assays, the biological activity of hyodeoxycholic acid is assessed in CHO cells stably expressing human TGR5. Cells are treated with varying concentrations of hyodeoxycholic acid (1-1000 microM), and intracellular cAMP levels are measured using a cAMP ELISA or a luminescence-based assay (e.g., LANCE Ultra cAMP kit). The EC50 for TGR5 activation is 31.6 microM. Hyodeoxycholic acid-d5 is used as an internal standard for quantifying drug exposure in these cells.
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| Enzyme Assay |
TGR5 activation assays are conducted using a cell-free membrane-based assay or a cellular cAMP assay. For cell-free receptor binding assays, membranes from TGR5-expressing CHO cells are incubated with a fluorescent or radiolabeled bile acid tracer in the presence of varying concentrations of hyodeoxycholic acid. Bound tracer is measured after filtration. For cellular cAMP assays, CHO-TGR5 cells are treated with hyodeoxycholic acid, and cAMP is quantified by ELISA. Hyodeoxycholic acid-d5 is used as an internal standard for mass spec validation.
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| Cell Assay |
CHO cells stably transfected with human TGR5 are seeded in 96-well white plates at 50,000 cells/well in Ham‘s F-12 medium. After 24 hours, cells are treated with hyodeoxycholic acid (0.1-1000 microM) in the presence of IBMX (phosphodiesterase inhibitor) for 30 minutes. Cells are lysed, and cAMP levels are measured using a homogeneous time-resolved fluorescence (HTRF) or luminescence-based cAMP assay kit. The EC50 is calculated by non-linear regression analysis. Parallel samples are analyzed by LC-MS/MS using HDCA-d5 as internal standard.
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| Animal Protocol |
Hyodeoxycholic acid-d5 is not commonly used in animal studies as a tracer. However, studies of unlabeled hyodeoxycholic acid can be conducted in LDLR-knockout mice to assess its effect on HDL function and atherosclerotic lesion formation. Hyodeoxycholic acid is administered orally or intraperitoneally, and bile acid levels in plasma and tissues are quantified by LC-MS/MS using HDCA-d5 as the internal standard. TGR5 activation can be inferred from downstream metabolic changes.
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| ADME/Pharmacokinetics |
Hyodeoxycholic acid-d5 has a molecular weight of 397.6 Da and is typically stored as a powder at -20degC. The deuterium label does not alter the physicochemical properties of the molecule. The compound serves as an internal standard for the accurate quantification of HDCA in LC-MS/MS assays. It is stable in biological matrices under standard storage conditions. In vivo pharmacokinetics of HDCA are similar to those of other bile acids, with enterohepatic circulation and a half-life determined by the rate of intestinal reabsorption and hepatic clearance.
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| Toxicity/Toxicokinetics |
Hyodeoxycholic acid-d5 is used in minute quantities as an analytical standard and does not contribute to toxicity. The unlabeled hyodeoxycholic acid has low toxicity; high doses may cause mild diarrhea or gastrointestinal discomfort due to its bile acid properties. In preclinical studies, hyodeoxycholic acid has been shown to improve HDL function and inhibit atherosclerotic lesion formation without significant adverse effects. No genotoxicity or carcinogenicity data are available.
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| References |
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| Additional Infomation |
Hyodeoxycholic acid is a naturally occurring secondary bile acid with documented TGR5 agonist activity. It has been investigated for its potential to improve HDL function and prevent atherosclerosis. The d5-labeled version is a research-grade isotopic standard used exclusively for analytical method development (LC-MS/MS) to quantify HDCA in biological fluids, tissue samples, and food matrices. It is not intended for therapeutic use.
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| Molecular Formula |
C24H35D5O4
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| Molecular Weight |
397.60
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| Related CAS # |
Hyodeoxycholic acid;83-49-8
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| Appearance |
Typically exists as solid at room temperature
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| 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 | 2.5151 mL | 12.5755 mL | 25.1509 mL | |
| 5 mM | 0.5030 mL | 2.5151 mL | 5.0302 mL | |
| 10 mM | 0.2515 mL | 1.2575 mL | 2.5151 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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