| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
Pregnenolone-d6 has no therapeutic target; it is an analytical standard. The unlabeled parent compound, pregnenolone, is a potent neurosteroid that acts as a signaling-specific inhibitor of the cannabinoid CB1 receptor. It is also a positive allosteric modulator (PAM) of the TRPM3 channel and a weak activator of TRPM1 channels. Pregnenolone is the precursor to all steroid hormones. As an internal standard, Pregnenolone-d6 is chemically identical to the endogenous analyte but is heavier due to the 6 deuterium atoms. This allows it to be distinguished by mass spectrometry (mass shift of +6 Da).
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| ln Vitro |
Stable heavy isotopes of hydrogen, carbon, and other elements have been incorporated into drug molecules, largely as tracers for quantitation during the drug development process. Studies involving the human use of drugs labeled with deuterium suggest that these compounds may offer some advantages when compared with their nondeuterated counterparts. Deuteration has gained attention because of its potential to affect the pharmacokinetic and metabolic profiles of drugs. Deutetrabenazine is the first deuterated drug to receive Food and Drug Administration approval. This deuterated form of the drug tetrabenazine is indicated for the treatment of chorea associated with Huntington's disease as well as tardive dyskinesia. Ongoing clinical trials suggest that a number of other deuterated compounds are being evaluated for the treatment of human diseases and not merely as research tools.
Pregnenolone-d6 itself is not biologically active. The unlabeled compound, pregnenolone, has been shown to inhibit the effects of tetrahydrocannabinol (THC) by blocking the CB1 receptor, thereby protecting the brain from cannabis intoxication. It modulates GABA-A and NMDA receptors. It activates TRPM3 channels, leading to calcium influx. The labeled version (d6) is used for quantitative bioanalysis. It has the same chemical stability and mass spectrometry fragmentation pattern as the unlabeled compound. |
| ln Vivo |
Deuterated compounds may, in some cases, offer advantages over nondeuterated forms, often through alterations in clearance. Deuteration may also redirect metabolic pathways in directions that reduce toxicities. The approval of additional deuterated compounds may soon follow. Clinicians will need to be familiar with the dosing, efficacy, potential side effects, and unique metabolic profiles of these new entities.
Pregnenolone-d6 is not administered in vivo. The endogenous parent compound, pregnenolone, is synthesized in the adrenal glands, gonads, and brain. Its concentration in biological fluids is a biomarker of adrenal and gonadal function. The labeled standard is added to patient samples (plasma, urine) to quantify endogenous pregnenolone levels. This is used in the diagnosis of adrenal insufficiency and congenital adrenal hyperplasia (CAH). It is not a therapeutic agent. |
| Enzyme Assay |
Pregnenolone-d6 is not a substrate for enzyme assays; it is an analytical standard. For the quantification of pregnenolone in human plasma, an LC-MS/MS method is used. A 200 uL plasma sample is spiked with 10 uL of Pregnenolone-d6 (internal standard, 100 ng/mL). The sample is extracted by liquid-liquid extraction with methyl tert-butyl ether (MTBE). The organic phase is evaporated and reconstituted in mobile phase. The extract is injected onto a C18 reverse-phase column (2.1 × 50 mm, 1.7 um). The mobile phase is water/methanol with 0.1% formic acid. Detection is performed in positive ion mode using multiple reaction monitoring (MRM) transition m/z 317 → 159 (for pregnenolone) and m/z 323 → 165 (for the d6 internal standard).
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| Cell Assay |
Pregnenolone-d6 is not used in cell-based assays. For toxicity screening, HepG2 or HEK293 cells are seeded in 96-well plates (1×10⁴ cells/well) and treated with Pregnenolone-d6 (1-1000 uM) for 24-72 h. Cell viability is measured by MTT assay. The CC₅0 is expected to be > 100 uM, indicating low cytotoxicity. For neurosteroid pharmacology, the unlabeled pregnenolone is used. The internal standard is only used in the analytical chemistry workflow.
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| Animal Protocol |
Pregnenolone-d6 is not a drug; it is an internal standard. For the purpose of validating an LC-MS/MS method for pregnenolone, blank rat plasma (200 uL) is spiked with known concentrations of unlabeled pregnenolone (0.1-100 ng/mL) to create a calibration curve. A fixed concentration of Pregnenolone-d6 (10 ng/mL) is added to all calibration standards, quality control (QC) samples, and study samples. The calibration curve is constructed by plotting the peak area ratio (analyte/IS) vs. the concentration. The method is validated for accuracy, precision, linearity, selectivity, and stability (i.e., freeze-thaw, bench-top).
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| ADME/Pharmacokinetics |
Pregnenolone-d6 is not a drug. It is a stable isotope-labeled standard. The six deuterium atoms are incorporated at specific positions (likely the C17 and C21 side chain). The mass difference of 6 Da (from 316.48 to 322.48) is sufficient to resolve the analyte from the internal standard in a mass spectrometer. The compound is stored at -20degC, protected from light and moisture. It is supplied as a white to off-white solid. The isotopic purity is > 99%.
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| Toxicity/Toxicokinetics |
For Pregnenolone-d6, hazard statements: H315 (Causes skin irritation), H319 (Causes serious eye irritation), H335 (May cause respiratory irritation). Signal word: Warning. Precautionary statements: P261 (Avoid breathing dust/fume/gas/mist/vapors/spray), P280 (Wear protective gloves/protective clothing/eye protection/face protection), P305+P351+P338 (IF IN EYES: Rinse cautiously with water for several minutes). Storage: at -20degC, in a tightly sealed container, protected from light. For research use only.
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| References |
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| Additional Infomation |
Pregnenolone-d6 (3beta-Hydroxy-5-pregnen-20-one-d6; CAS# 3599-13-5) is a research-grade stable isotope-labeled internal standard. It is not an FDA-approved drug. It is used for the quantification of the neurosteroid pregnenolone in biological samples by LC-MS/MS for steroid profiling, endocrinology, and neurochemistry research. For research use only, not for diagnostic or therapeutic applications.
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| Molecular Formula |
C21H26D6O2
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| Molecular Weight |
322.51
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| Related CAS # |
Pregnenolone; 145-13-1
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| Appearance |
Typically exists as solids at room temperature
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| Synonyms |
pregnenolone-d6; 3β-Hydroxy-5-pregnen-20-one-d6
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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 | 3.1007 mL | 15.5034 mL | 31.0068 mL | |
| 5 mM | 0.6201 mL | 3.1007 mL | 6.2014 mL | |
| 10 mM | 0.3101 mL | 1.5503 mL | 3.1007 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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