Estradiol-d4 - 10mM in DMSO , CAS No.66789-03-5

CAS: 66789-03-5 Cat. No.: E655792 Fórmula: C₁₈H₂₀D₄O₂ Peso molecular: 276.41

GRADE & PURITY 10mM in DMSO

Storage

Store at -80°C

Shipped In

Dry ice packs + Cold packs

Descripción general

Estradiol-d 4 is the deuterium labeled Estradiol. Estradiol is a steroid sex hormone vital to the maintenance of fertility and secondary sexual characteristics in females. Estradiol upregulates IL-6 expression through the estrogen receptor β (ERβ) pathway.

In 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. Deuteration has gained attention because of its potential to affect the pharmacokinetic and metabolic profiles of drugs. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Specifications

Especificaciones y pureza

10mM in DMSO

Mecanismos bioquímicos y fisiológicos

Estradiol-d 4 is the deuterium labeled Estradiol. Estradiol is a steroid sex hormone vital to the maintenance of fertility and secondary sexual characteristics in females. Estradiol upregulates IL-6 expression through the estrogen receptor β (ERβ) pathway

Condiciones de almacenamiento de almacenamiento

Store at -80°C

Enviado en

Dry ice packs + Cold packs

Este producto requiere envío en cadena de frío. Los servicios terrestres y otros servicios económicos no están disponibles.

Tipo de acción

AGONIST

Nombres e identificadores

Isómeros SMILES

[2H]C1=CC2=C(CC[C@@H]3[C@@H]2CC[C@]4([C@H]3CC([C@@H]4O)([2H])[2H])C)C(=C1O)[2H]

Peso molecular

276.41

Reaxy-Rn

2055276

Reaxys-RN_link_address

https://www.reaxys.com/reaxys/secured/hopinto.do?context=S&query=IDE.XRN=2055276&ln=

Application Protocols

The following are general, literature‑based protocols for using estradiol‑d4 as an internal standard in research LC–MS(/MS). They are provided for guidance only and are not item‑specific specifications.

• Stock and working solutions:
  • Prepare a primary stock at 0.5–1.0 mg/mL in methanol or acetonitrile. Sonicate briefly if needed. Protect from light.
  • Prepare serial working solutions (e.g., 1, 10, 100 ng/mL) in 50–80% methanol/water or neat methanol for spiking.
• Sample spiking (matrix‑matched calibration):
  • Add a fixed volume of working IS solution to each sample, calibrator, and QC prior to extraction to achieve a constant final IS concentration (e.g., 0.5–5 ng/mL depending on sensitivity requirements).
  • Vortex thoroughly; allow 5–10 min equilibration for binding/partitioning.
• Extraction (example, SPE):
  • Condition cartridge (MeOH → water). Load sample. Wash with water and 5–10% MeOH. Elute with MeOH or MeOH/ACN.
  • Evaporate at ≤40 °C under N2; reconstitute in initial LC mobile phase (e.g., 30–50% MeOH in water) containing a small, known IS amount if post‑extraction spiking is required.
• LC–MS/MS acquisition:
  • Monitor transitions for estradiol and estradiol‑d4 (analyte +4 m/z). Optimize collision energies to yield robust qualifier/quantifier ions.
• Adsorption control:
  • Use silanized glass vials; pre‑rinse autosampler needle and loop with standard solution.

Document recovery, matrix effects, linearity, and LOD/LOQ under your specific conditions using this IS.

Biological Roles

This section summarizes general, literature‑based biology of estradiol (17β‑estradiol); isotopic labeling with deuterium does not alter the intrinsic biological role but facilitates tracing and quantification in research. No medical or clinical claims are made.

• Endogenous ligand: principal estrogen in many mammals, produced mainly by ovarian granulosa cells and peripheral aromatization pathways.
• Receptor interactions: binds nuclear estrogen receptors ERα and ERβ, as well as membrane‑associated receptors (e.g., GPER1), initiating genomic and non‑genomic signaling cascades (literature).
• Physiological roles (high‑level): regulation of reproductive tissue development, secondary sex characteristics, bone turnover, lipid metabolism, and neuroendocrine signaling (literature).
• Biotransformation: subject to oxidative metabolism (e.g., 2‑ and 4‑hydroxylation), interconversion with estrone, and phase‑II conjugation (sulfation, glucuronidation). These pathways are often monitored using deuterated tracers to follow turnover and compartmentalization in research systems.
• Transport and distribution: circulates bound to sex hormone‑binding globulin and albumin; free fraction is bioactive (literature). In vitro, nonspecific binding to plastics and glass can be significant—account for this when designing dosing experiments.
• Analytical relevance of deuteration: deuterium label provides a mass offset without materially affecting receptor affinity or partitioning at typical labeling sites, enabling precise isotope‑dilution mass spectrometry of endogenous estradiol in research matrices.

Note: For any work involving cells or organisms, ensure compliance with institutional biosafety and ethical guidelines; use this material strictly for research purposes as labeled by the supplier.

Buffer Applications

Estradiol‑d4 is not a buffering reagent and does not form a defined buffer system. It is a hydrophobic, weakly acidic phenolic compound used as an analytical internal standard.

• Practical note: When preparing solutions in aqueous buffers, dissolve first in a miscible organic solvent (e.g., methanol, acetonitrile, or DMSO), then dilute into the buffer with vigorous mixing to avoid precipitation. Keep final organic content compatible with your biological or analytical system.

For true buffering needs, select dedicated buffer components (e.g., phosphate, Tris, HEPES) appropriate to the target pH range.

Green Alternatives

While estradiol‑d4 itself has no direct “green” substitute for isotope‑dilution analytics, the surrounding workflow can often be improved using greener choices without compromising data quality.

Greener solvent choices (literature guidance):

• Replace chlorinated solvents (e.g., dichloromethane in liquid–liquid extraction) with:
  • Ethyl acetate or methyl tert‑butyl ether (MTBE) for non‑aqueous extractions.
  • Heptane/iso‑octane for nonpolar phases when appropriate.
• Favor bio‑based and lower‑toxicity alcohols:
  • Ethanol over methanol for stock solutions or SPE conditioning when compatible with the LC–MS method (be mindful of slightly different elution strengths and ionization).
• Optimize LC mobile phases:
  • Use water/ethanol or water/methanol in place of acetonitrile where chromatographic performance allows; adjust gradient and temperature to compensate for viscosity.
• Solid‑phase extraction (SPE) over LLE:
  • Reduces solvent volumes; polymeric reversed‑phase sorbents can provide high recoveries for phenolic steroids with greener wash/elution protocols.

Trade‑offs and controls:

• Ethanol’s higher viscosity and UV cutoff may affect backpressure and detection; re‑optimize gradients and detector settings.
• MTBE is more volatile and flammable; ensure ventilation and ignition control.
• Any solvent change must be validated for recovery, matrix effects, and ion suppression using the same estradiol‑d4 internal standard to maintain data integrity.

Conclusion: Keep the deuterated standard as designed, but redesign extraction and chromatography with safer, lower‑impact solvents where performance permits.

Pharmaceutical Uses

No therapeutic or clinical claims are made for this product. In a research and quality‑control context, deuterated estradiol standards are commonly used as:

• Analytical reference standards and internal standards for method development and validation in pharmaceutical analytics (e.g., assay of estradiol content in research samples or formulations by LC–MS or GC–MS after derivatization; literature practice).
• System suitability and calibration controls to assess instrument response, extraction efficiency, and matrix effects in research workflows.

Excipient status/pharmacopeia monographs: Not applicable to the deuterated standard; estradiol (unlabeled) may have pharmacopeial monographs, but those do not govern this deuterated research reagent. Always treat this product strictly as a research chemical, per the supplier’s “For research use only” designation.

Physical Properties

Item-specific physicochemical specifications are not provided for this lot. The following values are general literature information for estradiol or deuterated estradiol and should be treated as guidance only (not product specifications).

• Appearance: Not specified for this item; refer to CoA/Spec Sheet. (Literature: off‑white crystalline solid for non‑deuterated estradiol)
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet. (Literature: estradiol 272.38 g/mol; d4 analog typically ~276.4 g/mol)
• Melting point (literature, estradiol): ~176–180 °C (polymorph/solvate dependent). Deuteration minimally affects MP.
• Boiling point: Not typically defined (decomposes before boiling under ambient pressure; literature).
• Density: Not specified for this item; refer to CoA/Spec Sheet.
• pKa (literature, estradiol): phenolic OH ~10.3–10.7; secondary alcohol pKa much higher (non‑ionizing under neutral conditions). Isotopic substitution has negligible effect on acidity.
• LogP (literature, estradiol): ~3.8–4.1; high lipophilicity consistent with steroidal scaffold.
• Aqueous solubility (literature): very low in water (µg/mL); readily soluble in polar aprotic/protic organics (DMSO, DMF, EtOH, MeOH, acetone). Solubility of d4 closely matches unlabeled.
• UV/Vis (literature, estradiol): phenolic A‑ring gives λmax ~280 nm (MeOH), with weaker bands around 210–230 nm.
• Refractive index/RI and specific rotation: Not applicable to solids or not commonly reported; Not specified for this item.

Note: Deuteration does not materially change bulk physical properties vs. protiated estradiol; mass spectrometric behavior differs (m/z shift +4, slightly altered fragmentation pathways). Always verify any property critical to your method with the item’s CoA.

Quality and Grades

• Item-specific grade/purity: Not specified for this item; refer to CoA/Spec Sheet.

Context for professional users working with deuterated small‑molecule standards:

• Isotopic enrichment: For quantitative LC–MS(/MS), high deuterium enrichment (often ≥98 atom % D at labeled positions) is typically sought to ensure a distinct +4 m/z channel with minimal isotopic cross‑talk. The actual enrichment for this item is Not specified; confirm on the CoA.
• Positional labeling: The exact locations of the four deuterons affect metabolic stability (e.g., resistance to back‑exchange) and fragmentation patterns. CoA typically discloses the labeling scheme and any residual unlabeled isotopologues.
• Chemical purity: Analytical/reference‑standard grades often target ≥95–98% chemical purity with tight controls on isomers and residual solvents. For chromatographic work, low levels of UV‑absorbing impurities are important. This item’s purity is Not specified; verify on the CoA.
• Residual solvents, water content, metal content, and stabilizers: Not specified for this item; refer to CoA/Spec Sheet.
• Why grade matters:
  • Quantitation accuracy: isotopic and chemical purity directly influence internal‑standard response factors and matrix‑matched calibration.
  • Stability: micro‑impurities can catalyze oxidation of phenolics; suppliers may package under inert atmosphere or with desiccants.
• Acceptance testing suggestions:
  • Verify m/z spacing and absence of interfering isotopologues by direct infusion or LC–MS.
  • Check retention match with native estradiol under your LC method.
  • Run purity checks by HPLC/UPLC with photodiode array and MS detection.

For regulated work, retain the CoA, lot traceability, and, if needed, perform in‑house qualification under your method.

Reaction and Applications

Primary value of estradiol‑d4 is analytical rather than synthetic. Key research applications include:

• Isotope‑dilution quantification (LC–MS/MS; GC–MS after derivatization):
  • Spiked as an internal standard to correct for extraction efficiency, ionization variability, and matrix effects when quantifying native estradiol in research samples.
  • The +4 m/z shift provides a clear mass channel with near‑identical chromatographic behavior to the analyte (literature best practice).
• Metabolic and mechanistic studies:
  • Tracing biotransformation pathways (oxidation, conjugation) via MS by following the fate of deuteriums; positional labeling informs hydrogen migration or exchange events (literature).
  • Kinetic isotope effect (KIE) probes at specific C–H sites to assess rate‑limiting steps in model reactions catalyzed by enzymes or synthetic catalysts.
• Derivatization workflows (analytical):
  • GC–MS: trimethylsilylation of the 3‑ and 17‑OH groups (e.g., MSTFA, BSTFA, or MTBSTFA) to improve volatility/thermal stability.
  • LC–MS or LC–FLD: pre‑column dansylation of the phenolic OH to enhance ionization/fluorescence (literature).
• Reference standard and system suitability:
  • Calibration curves, retention time markers, and MS tuning (product‑independent practice).

Practical tips:

• Maintain consistent organic content in spike solutions; typical working levels are in the ng/mL–µg/mL range depending on matrix and detector.
• Protect from light; use amber vials and minimize autosampler residence times to reduce adsorption and slow oxidation.
• Verify absence of H/D back‑exchange under derivatization or high‑pH conditions if positional integrity matters (literature caution).

Reaction Conditions

Typical literature conditions related to analytical derivatization and sample preparation for estradiol/estradiol‑d4 (general guidance; not product specifications):

• GC–MS silylation:
  • Reagent: MSTFA (often with 1% TMCS) or BSTFA.
  • Conditions: 60–70 °C for 20–60 min in anhydrous solvent (acetonitrile or pyridine). Dry sample thoroughly to avoid hydrolysis of silyl reagents. Produces bis‑TMS derivatives for improved volatility.
• LC–MS dansylation (phenolic tagging):
  • Reagent: dansyl chloride (DNS‑Cl).
  • Conditions: carbonate/bicarbonate buffer (pH ~9–10) in acetonitrile/water; 40–60 °C for 10–30 min. Quench excess reagent; inject onto reversed‑phase LC. Enhances sensitivity in ESI+.
• SPE cleanup for biological matrices (research use):
  • Sorbent: polymeric reversed‑phase (e.g., divinylbenzene‑based) or C18.
  • Conditioning: MeOH → water.
  • Load: aqueous sample (≤10% organic).
  • Wash: water, then weak organic.
  • Elute: MeOH or MeOH/ACN.
• LC conditions (common starting points):
  • Column: C18 or phenyl‑hexyl, 2.1 × 100 mm, 1.7–3 µm.
  • Mobile phase A: water + 0.1% formic acid (or ammonium formate 2–10 mM).
  • Mobile phase B: methanol or acetonitrile.
  • Gradient: 40→90% B over 5–8 min; 30–50 °C column temp; flow 0.2–0.4 mL/min.
  • MS: ESI+ or APCI+; monitor MRM transitions shifted by +4 m/z relative to estradiol (method‑specific).

Notes:

• Verify that derivatization does not induce H/D exchange at labeled positions of interest.
• Always re‑optimize for your instrument and matrix; the above are literature‑based starting parameters.

Safety and Handling

Safety classifications for this specific item are not provided here; consult the SDS for authoritative information. The following are prudent, chemistry‑focused handling practices for deuterated estradiol standards.

• GHS/Signal word/pictograms: Not specified for this item; refer to SDS. Estradiol (unlabeled) is an endocrine‑active steroid and may carry hazard statements related to reproductive toxicity in some jurisdictions (literature); treat with heightened caution.
• Personal protective equipment (typical lab practice):
  • Lab coat, nitrile gloves (double‑glove for prolonged handling), chemical splash goggles.
  • Handle powders/solids in a certified fume hood or ventilated balance enclosure to prevent inhalation and cross‑contamination.
• Avoid: skin contact, inhalation of dust/aerosols, and accidental ingestion. Prevent environmental release; adsorbs strongly to labware.
• Storage incompatibilities: strong oxidizers, strong bases (may promote phenolate formation and side reactions), and reactive acylating/silylating agents unless intentionally derivatizing.
• Special risks and precautions:
  • Endocrine activity: even trace exposure may be significant; maintain closed systems for solution prep and spiking.
  • Sorption/adsorption: steroids can adsorb to glass and plastics; pre‑rinse with solvent or use silanized glass/low‑bind plastics.
  • Light and air: protect from light; keep solutions dry and oxygen‑limited to minimize slow oxidative changes.
• First‑aid (overview; follow institutional protocols):
  • Skin/eye contact: immediate decontamination with copious water; remove contaminated clothing; seek medical evaluation.
  • Inhalation: move to fresh air; monitor; seek medical evaluation.
  • Ingestion: rinse mouth, do not induce vomiting; seek medical attention.

Always defer to and follow the item‑specific SDS and your institution’s EHS guidance.

Solvent Selection

Estradiol‑d4 is a lipophilic, weakly acidic phenolic steroid. Solvent choice should balance solubility, adsorption control, and compatibility with the analytical workflow.

• Polarity/solubility profile (literature):
  • Highly soluble: DMSO, DMF, acetone, acetonitrile (moderate–good), methanol, ethanol, isopropanol.
  • Poorly soluble: water and aqueous buffers without cosolvent; solubility in pure water is typically in the low µg/mL range.
• Recommended stock solvents for standards:
  • Methanol or acetonitrile for LC–MS workflows (volatile, MS‑compatible). DMSO for high‑concentration master stocks with subsequent dilution into MeOH/ACN.
• Adsorption mitigation:
  • Pre‑condition vials and syringes with 1–2 rinses of the standard solution.
  • Prefer silanized glass or low‑bind polypropylene; avoid soda‑lime glass for trace‑level work.
• Mobile phase compatibility:
  • Reversed‑phase LC: water/MeOH or water/ACN with 0.1% formic acid or ammonium formate typically used; d4 co‑elutes closely with native estradiol—fine‑tune gradient or use phenyl‑hexyl phases for subtle selectivity shifts (literature guidance).
• Sample preparation:
  • For aqueous matrices, employ ≥40–80% organic cosolvent for spiking to avoid precipitation; keep final organic ≤5–10% for biological extractions (research use).

Brief comparison (literature):

• Methanol: strong solvation for phenolics; promotes reproducible injection, slightly higher background in some ESI methods.
• Acetonitrile: lower viscosity, sharper peaks; slightly lower solubility than MeOH for some steroids.
• DMSO: maximum solubility; limited injection volumes due to ESI suppression—dilute into MeOH/ACN before LC–MS.

Storage and Reconstitution

• Storage (item‑specific):
  • Store at −80 °C. Protect from light and moisture. Supplier ships on dry ice packs + cold packs, consistent with deep‑freeze storage requirements.
• Stability: Not specified for this item; refer to CoA/Spec Sheet. General practice: keep solid tightly capped under inert atmosphere or desiccated. Minimize temperature cycling.
• Reconstitution (general guidance for research use):
  • Allow vial to equilibrate to room temperature in a desiccator before opening to prevent moisture condensation.
  • Dissolve in dry methanol, acetonitrile, or DMSO to prepare a concentrated stock (e.g., 0.5–1.0 mg/mL). Gently vortex/sonicate if needed. Protect from light.
  • Filter not required for neat organic stocks; for aqueous working solutions, 0.2 µm PTFE filtration may be used if particulate risk exists.
• Aliquoting and freeze–thaw:
  • Prepare small single‑use aliquots to avoid repeated freeze–thaw. Store aliquots in amber, low‑bind vials at −80 °C.
  • Avoid adsorptive losses by pre‑rinsing vials with a small volume of the stock solution when preparing aliquots.
• Shipping:
  • Shipped on dry ice packs + cold packs as provided. Inspect upon receipt; if thawing occurred, consult CoA/SDS and evaluate integrity by LC–MS before use.

Always defer to the product’s CoA and SDS for definitive handling, stability, and storage instructions. Research use only.

Structure and Identity

Estradiol‑d4 is a deuterium-labeled analog of 17β‑estradiol used predominantly as an isotope internal standard and tracer in analytical chemistry and biochemistry.

• Product identifiers (item-specific):

  • SKU: E655792
  • Name: Estradiol‑d4
  • CAS: 66789-03-5
  • PubChem CID: 87229045
  • InChIKey: Not specified for this item; refer to CoA/Spec Sheet.
  • SMILES: Not specified for this item; refer to CoA/Spec Sheet.

• Molecular descriptors:

  • Molecular formula: Not specified for this item; refer to CoA/Spec Sheet. (Note: non‑deuterated estradiol is C18H24O2; d4 isotopologue replaces four 1H with 2H; literature)
  • Molecular weight: Not specified for this item; refer to CoA/Spec Sheet. (Note: estradiol is ~272.38 g/mol; d4 isotopologue typically ~276.4 g/mol; literature)

• Structural features (general, literature):

  • Core: fused tetracyclic estrane (steroid) framework: three six‑membered rings (A, B, C) and one five‑membered ring (D).
  • Functional groups: two phenolic/aliphatic hydroxyls (A‑ring phenol at C3; secondary alcohol at C17β).
  • Aromaticity: A‑ring is phenolic/aryl; B–D rings are saturated.
  • Stereochemistry: defined steroidal stereocenters across the C/D rings; biological form is 17β‑hydroxy orientation.
  • Isotopic labeling: four deuterons incorporated at supplier‑specified positions (common literature patterns include 2,4,16,16‑d4 or 6,7,16,16‑d4). Exact labeling pattern for this item is Not specified; refer to CoA/Spec Sheet.

• 2D description (general): a planar phenolic A‑ring bearing a para‑fused cyclohexane B‑ring leading into chair cyclohexane C‑ring and a puckered cyclopentane D‑ring with a C17β‑OH. Deuteriums substitute specific ring hydrogens without altering the carbon scaffold.

Synthetic Utility

Estradiol‑d4 is typically not used as a building block for organic synthesis; its primary value lies in isotopic labeling for analytics. That said, its functional groups enable certain transformations for method development and mechanistic studies (literature guidance):

• Phenolic and secondary alcohol reactivity:
  • O‑silylation (TMS, TBDMS, TBS) for GC–MS volatility or to modulate polarity in LC.
  • O‑acylation or O‑carbamoylation (e.g., dansyl chloride, benzoyl chloride) to enhance detectability via UV/FLD/ESI.
• Oxidation/reduction interconversions:
  • Controlled oxidation at C17 (estradiol → estrone) or reduction (estrone → estradiol) to study isotope scrambling or retention of deuterium at ring positions.
• Electrophilic aromatic substitution (EAS) on the phenolic A‑ring (e.g., iodination/nitration under mild conditions) as probes for regioselectivity or to tune MS ionization; deuterium kinetic isotope effects can be probed if labeling resides on reactive positions.
• Conjugation chemistry:
  • Formation of sulfate or glucuronide conjugates in vitro (enzymatic or chemical) to map isotope transfer and monitor conjugate stability by MS.

Caveats:

• H/D back‑exchange: Avoid strong base, acidic D2O, or high temperatures if positional integrity of deuterium is essential; aliphatic C–D bonds are generally robust, while phenolic O–H is protiated unless specifically deuterated.
• Use the deuterated analog sparingly in synthetic steps; reserve for final derivatization or analytical calibration to minimize isotopic dilution.

Target Specificity

Not applicable. This product is a small‑molecule deuterated steroid standard, not a biological affinity reagent (e.g., antibody or ligand with defined target specificity). For receptor binding studies in research, consult literature on estradiol interactions with ERα/ERβ/GPER1, but such specificity is not a property of this catalog item itself.