| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Targets |
Mono-Cyclohexyl Phthalate-3,4,5,6-d4 is a stable isotope-labeled internal standard. Its unlabeled parent, mono-cyclohexyl phthalate (MCHP), is not a drug but a metabolite of the plasticizer dicyclohexyl phthalate (DCHP). MCHP is considered a biomarker of human exposure to DCHP. In toxicological studies, MCHP has been identified as an endocrine-disrupting chemical (EDC). It has been shown to activate the nuclear receptor peroxisome proliferator-activated receptor gamma (PPARgamma) and to antagonize the androgen receptor (AR). MCHP also acts as a weak agonist of the estrogen receptor (ER). These activities are consistent with the antiandrogenic and obesogenic effects attributed to some phthalates. The primary molecular targets of MCHP (and its parent DCHP) are believed to be: (1) PPARgamma, which plays a role in adipogenesis (fat cell formation) and glucose homeostasis, and (2) steroidogenic enzymes (e.g., aromatase, 17beta-hydroxysteroid dehydrogenase). The labeled version is used as an internal standard to accurately quantify MCHP in exposure assessment studies.
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| ln Vitro |
Drug compounds have included stable heavy isotopes of carbon, hydrogen, and other elements, mostly as quantitative tracers while the drugs were being developed. Because deuteration may have an effect on a drug's pharmacokinetics and metabolic properties, it is a cause for concern [1].
The in vitro biological activity of Mono-Cyclohexyl Phthalate-3,4,5,6-d4 is presumed to be identical to its unlabeled parent, MCHP. In cell-based reporter gene assays, MCHP (1-100 uM) has been shown to activate PPARgamma in a concentration-dependent manner, with an EC₅0 of approximately 10-30 uM. In the same concentration range, MCHP exhibits weak anti-androgenic activity: it inhibits the dihydrotestosterone (DHT)-induced activation of the androgen receptor (AR) in a reporter gene assay (IC₅0 ~ 20-50 uM). In the MCF-7 breast cancer cell line (estrogen receptor-positive), MCHP (10-100 uM) has been reported to weakly increase cell proliferation (estrogenic effect). In the H295R human adrenocortical carcinoma cell line, MCHP (10-100 uM) inhibits the activity of CYP19 aromatase (which converts androgens to estrogens) and alters the expression of steroidogenic enzymes, leading to decreased production of estradiol and testosterone. The labeled MCHP-d4 is used as an internal standard to accurately quantify MCHP concentrations in the cell culture medium in these in vitro studies, ensuring proper dose-response characterization. |
| ln Vivo |
The in vivo activity of Mono-Cyclohexyl Phthalate-3,4,5,6-d4 is not directly evaluated; it is used as an internal standard. Its unlabeled parent, MCHP, is the major urinary metabolite of dicyclohexyl phthalate (DCHP). In animal studies, oral administration of DCHP (50-500 mg/kg/day for 28 days) to male rats leads to high levels of MCHP in plasma and urine. DCHP exposure in rats has been associated with reproductive toxicity, including reduced testosterone production, decreased sperm count, and histopathological changes in the testis (Leydig cell hyperplasia, seminiferous tubule degeneration). These effects are thought to be due, at least in part, to the accumulation of MCHP and its activity as an anti-androgen. In pregnant rats, DCHP (100-1,000 mg/kg/day) crosses the placenta and produces MCHP in the fetal circulation, leading to developmental reproductive tract malformations in male offspring (reduced anogenital distance, retained nipples). MCHP-d4 is used as an internal standard to accurately quantify MCHP in maternal and fetal plasma and urine in these toxicological studies, enabling the establishment of dose-response relationships and the assessment of the internal dose of the toxicant.
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| Enzyme Assay |
A generic non-cell-based assay for Mono-Cyclohexyl Phthalate-3,4,5,6-d4 involves its use as an internal standard in an LC-MS/MS method for quantifying MCHP in human urine. Prepare a standard stock solution of unlabeled MCHP in methanol (1 mg/mL). Prepare a separate stock solution of the internal standard Mono-Cyclohexyl Phthalate-3,4,5,6-d4 at the same concentration. Prepare calibration standards by spiking the unlabeled analyte into a blank matrix (e.g., synthetic urine or control urine from a low-exposure individual) to achieve concentrations ranging from 0.1 to 100 ng/mL. Add a fixed concentration of the internal standard (e.g., 10 ng/mL) to each calibration standard. For sample preparation, add 10 uL of beta-glucuronidase (from E. coli, 1,000 units) to 1 mL of urine and incubate at 37degC for 12 hours to hydrolyze any conjugated MCHP (glucuronide). After hydrolysis, perform solid-phase extraction (SPE) using a mixed-mode anion exchange (MAX) cartridge. Load the urine sample, wash with 5% ammonium hydroxide, and elute with 2% formic acid in methanol. Evaporate the eluate under nitrogen and reconstitute in 200 uL of mobile phase. Analyze by LC-MS/MS in negative ion mode (or positive mode depending on ionization). Monitor the mass transitions: m/z 249 → 121 (loss of the cyclohexyl ring) for MCHP, and m/z 253 → 125 for the MCHP-d4 internal standard. Construct the calibration curve by plotting the peak area ratio (analyte/IS) vs. the nominal concentration. This method is used for human biomonitoring studies to measure phthalate exposure.
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| Cell Assay |
A standard in vitro cell-based protocol for the unlabeled MCHP involves the assessment of PPARgamma activation in the 3T3-L1 preadipocyte differentiation model. Culture 3T3-L1 mouse embryonic fibroblasts in DMEM supplemented with 10% FBS and 1% penicillin/streptomycin at 37degC in a 5% CO2 incubator. Seed cells in 12-well plates at 2×10⁵ cells/well and allow to reach confluence (Day -2). On Day 0 (two days post-confluence), initiate differentiation with differentiation medium (DMEM with 10% FBS, 0.5 mM IBMX, 1 uM dexamethasone, and 1 ug/mL insulin) plus increasing concentrations of unlabeled MCHP (0.1, 1, 10, 50, 100 uM) or the positive control rosiglitazone (1 uM, a PPARgamma agonist). On Day 2, change to the maintenance medium (DMEM with 10% FBS and 1 ug/mL insulin) plus the same MCHP concentrations. On Day 4, refresh the maintenance medium. On Day 6-8, assess adipocyte differentiation by (1) Oil Red O staining to visualize intracellular lipid droplets; (2) quantification of triglyceride content; or (3) measurement of the expression of adipogenic marker genes (aP2, PPARgamma, Adipsin) by qRT-PCR. Use the internal standard MCHP-d4 in the LC-MS analysis of the culture medium to confirm the exposure concentrations. This protocol is used to study the obesogenic potential of phthalate metabolites.
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| Animal Protocol |
A typical in vivo animal protocol for Mono-Cyclohexyl Phthalate-3,4,5,6-d4 involves a toxicokinetic study of dicyclohexyl phthalate (DCHP) in rats. Use male Sprague-Dawley rats (200-250 g, n = 5-6 per group). Administer a single oral dose of unlabeled DCHP (100 mg/kg) suspended in corn oil via gavage. Collect blood samples via tail vein at various time points (0, 0.5, 1, 2, 4, 6, 8, 12, 24, 36, 48 h) into heparinized tubes. Immediately centrifuge to obtain plasma. Also collect 24-hour urine samples using metabolic cages. For bioanalysis, hydrolyze the plasma and urine samples (with beta-glucuronidase) to release free MCHP from its conjugates. Add a fixed amount of Mono-Cyclohexyl Phthalate-3,4,5,6-d4 as the internal standard. Extract by SPE or liquid-liquid extraction (ethyl acetate). Analyze by LC-MS/MS. Quantify the concentration of MCHP in plasma and urine. Calculate the PK parameters (Cmax, Tmax, AUC, t½) for MCHP. Also, determine the cumulative urinary excretion of MCHP (as a percentage of the administered DCHP dose) over 48 hours. This protocol is used to assess the absorption, metabolism, and elimination kinetics of DCHP and its primary metabolite MCHP. The labeled internal standard ensures the accuracy of the quantification, which is critical for risk assessment of phthalate exposure.
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| ADME/Pharmacokinetics |
Mono-Cyclohexyl Phthalate-3,4,5,6-d4 is an analytical internal standard. Its unlabeled parent, MCHP, is a phthalate metabolite. The pharmacokinetics of MCHP in animals and humans are dependent on the PK of the parent diester (DCHP). DCHP is absorbed from the gastrointestinal tract, hydrolyzed to MCHP by esterases, and then MCHP is further metabolized (hydroxylation, oxidation) and conjugated (glucuronidation). The Tmax for MCHP in plasma is typically 1-3 hours after DCHP administration in rats. The half-life of MCHP in plasma is approximately 4-8 hours. In humans, after a single exposure to DCHP (e.g., from industrial sources or via the environment), MCHP is detectable in urine for up to 24-48 hours. MCHP is primarily eliminated in urine as the glucuronide conjugate. The concentration of MCHP in spot urine samples is used as a biomarker of recent DCHP exposure. The labeled internal standard is essential for the accurate quantification of this metabolite in human biomonitoring studies (e.g., NHANES) and in epidemiological studies investigating the health effects of phthalate exposure.
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| Toxicity/Toxicokinetics |
Mono-Cyclohexyl Phthalate-3,4,5,6-d4 is a research-grade stable isotope-labeled compound, not a pharmaceutical drug. Its unlabeled parent, MCHP, is a phthalate metabolite with endocrine-disrupting potential. Dicyclohexyl phthalate (DCHP) and its metabolite MCHP are classified as toxic to reproduction and suspected endocrine disruptors by regulatory agencies (e.g., ECHA, REACH). In animal studies, DCHP (and MCHP) have been shown to cause: (1) reproductive toxicity: reduced fertility, decreased sperm count, and testicular atrophy in male rats; (2) developmental toxicity: malformations of the male reproductive tract (reduced anogenital distance) in the offspring of pregnant rats exposed to DCHP; (3) liver toxicity: increased liver weight and hepatocellular hypertrophy at high doses. There is limited evidence of carcinogenicity. For laboratory handling, standard precautions for handling potentially hazardous chemicals should be used: wear gloves, a lab coat, and safety goggles; work in a fume hood; avoid generating dust; wash hands thoroughly. Mono-Cyclohexyl Phthalate-3,4,5,6-d4 should be stored as a powder at -20degC in a tightly sealed container, protected from light and moisture. It is for research use only, not for human consumption.
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| References | |
| Additional Infomation |
Mono-Cyclohexyl Phthalate-3,4,5,6-d4 is the stable isotope-labeled version of mono-cyclohexyl phthalate (MCHP), the primary urinary metabolite of the plasticizer dicyclohexyl phthalate (DCHP). DCHP is used as a plasticizer in polyvinyl chloride (PVC) formulations, cellulose acetate plastics, and in specialty applications such as lacquers, adhesives, and printing inks. Like other phthalates, DCHP is classified as an endocrine-disrupting chemical (EDC), with evidence of anti-androgenic and reproductive toxicity in animal studies. MCHP is a sensitive and specific biomarker of human exposure to DCHP. Human exposure to DCHP occurs through ingestion of contaminated food (migration from packaging materials), inhalation of indoor air (from PVC flooring and wall coverings), and dermal contact. Mono-Cyclohexyl Phthalate-3,4,5,6-d4 is intended for research use as an internal standard for the accurate quantification of MCHP in urine and plasma by LC-MS/MS. This labeled compound is an essential analytical tool for human biomonitoring studies, environmental epidemiology, and toxicological research on phthalates. For research use only, not for diagnostic or therapeutic applications.
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| Molecular Formula |
C14H12D4O4
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|---|---|
| Molecular Weight |
252.30
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| Exact Mass |
252.129
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| CAS # |
1398066-18-6
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| PubChem CID |
131698665
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| Appearance |
White to off-white solid powder
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| Density |
1.2±0.1 g/cm3
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| Boiling Point |
410.2±28.0 °C at 760 mmHg
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| Flash Point |
154.6±17.5 °C
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| Vapour Pressure |
0.0±1.0 mmHg at 25°C
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| Index of Refraction |
1.564
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| LogP |
3.19
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
18
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| Complexity |
301
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| Defined Atom Stereocenter Count |
0
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| SMILES |
[2H]C1=C(C(=C(C(=C1[2H])C(=O)[O-])C(=O)OC2CCCCC2)[2H])[2H]
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| InChi Key |
PMDKYLLIOLFQPO-DOGSKSIHSA-M
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| InChi Code |
InChI=1S/C14H16O4/c15-13(16)11-8-4-5-9-12(11)14(17)18-10-6-2-1-3-7-10/h4-5,8-10H,1-3,6-7H2,(H,15,16)/p-1/i4D,5D,8D,9D
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| Chemical Name |
2-cyclohexyloxycarbonyl-3,4,5,6-tetradeuteriobenzoate
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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 | 3.9635 mL | 19.8177 mL | 39.6354 mL | |
| 5 mM | 0.7927 mL | 3.9635 mL | 7.9271 mL | |
| 10 mM | 0.3964 mL | 1.9818 mL | 3.9635 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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