Mephenytoin-C,d , CAS No.1261398-53-1

CAS: 1261398-53-1 Cat. No.: M1438465 Formula: C₁₁₁₃CH₁₁D₃N₂O₂ Peso molecolare: 222.26

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Panoramica

Mephenytoin-13C,d3 is a deuterated labeled Mephenytoin. Mephenytoin, an anticonvulsant, is the CYP2C19 and CYP2B6 substrate.

Specifications

Condizioni di conservazione di stoccaggio

Store at -20°C

Spedito in

Ice chest + Ice pads

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Nomi e identificatori

Peso molecolare

222.26

Application Protocols

No manufacturer-validated protocols are provided in the Product Data. The following high-level workflows are common for isotopically labeled small-molecule standards and are offered as general guidance only.

• Preparation of calibration standards (LC–MS)

  • Prepare a primary stock in DMSO or methanol at a known concentration (e.g., 1–10 mM). Serially dilute with 50:50 acetonitrile:water (with 0.1% formic acid or ammonium buffer) to prepare a calibration series. Spike a constant amount of the isotopic standard into all calibrators, QCs, and samples.

• Internal standard spiking

  • Add the isotopic standard to biological matrices prior to extraction (protein precipitation or SPE) to correct for recovery and matrix effects. Typical internal standard concentration is chosen to bracket expected analyte levels and maximize precision.

• In vitro metabolism assay

  • Incubate substrate with microsomes/recombinant enzyme as outlined in Reaction Conditions. Quench with organic solvent containing the isotopic standard (if not pre-spiked). Analyze supernatants by LC–MS/MS.

• Documentation

  • Record lot number, isotopic enrichment, and preparation details in your method records; retain the CoA for audits/validation.

Adjust solvent systems, concentrations, and detection parameters to your instrument and matrix. Always validate before routine use.

Biological Roles

Item-specific biological properties are not provided in the Product Data. The following context pertains to unlabeled mephenytoin and is offered for research planning only (not a clinical claim).

• Context (literature/general)

  • Mephenytoin is a hydantoin-class compound that serves as a well-known probe substrate for cytochrome P450 2C19 (CYP2C19) in biochemical assays. Its metabolic conversion (e.g., aromatic hydroxylation to 4′-hydroxymephenytoin) is commonly used to assess CYP2C19 catalytic activity in vitro.
  • Isotopically labeled analogs (13C/2H) are inert tracers that mirror the biochemical behavior of the unlabeled compound, enabling accurate mass spectrometric quantification of parent and metabolites without altering enzyme kinetics materially.

• Utility in systems biology/ADME research

  • Serves as a benchmark substrate in comparative enzyme phenotyping across microsomal preparations or recombinant enzymes.
  • Facilitates determination of intrinsic clearance, formation rates of specific metabolites, and cofactor dependencies (e.g., NADPH in microsomal assays) when paired with stable-isotope internal standards.

• Limitations

  • In vitro use only as a research tool; not intended for clinical diagnostics or therapeutic applications.
  • Isotope effects on chromatography are typically minimal for 13C labeling; deuterium labeling can cause small retention shifts (secondary isotope effects) and should be considered in method development.

Buffer Applications

This product is not a buffering agent and is not typically used to prepare buffer systems. For experiments involving enzyme incubations or LC–MS analyses, select buffers appropriate to your platform:

• In vitro metabolism assays (general guidance)

  • Phosphate or Tris buffers (50–100 mM) at pH 7.4 are commonly used with liver microsomes; include MgCl2 as required; maintain ionic strength compatible with enzyme activity.
  • Use volatile buffers (ammonium formate/acetate, 2–20 mM) for LC–MS compatibility when direct injection of incubation matrices is planned.

• Practical notes

  • Limit organic cosolvent to minimize enzyme inhibition.
  • Filter or centrifuge samples prior to LC–MS to prevent salt buildup in the source.

Item-specific buffer capacity or pKa data are not applicable; refer instead to Solvent Selection and Reaction Conditions for use-context details.

Green Alternatives

As an isotopically labeled reference standard, this product has no direct “greener substitute” for its core analytical role. However, greener practices can be applied to its use and to solvent selection in associated workflows.

• Greener handling strategies (general)

  • Use minimal solvent volumes and prepare concentrated stocks (e.g., in DMSO) to reduce waste; dilute at point-of-use.
  • Prefer ethanol or water-miscible greener solvents when compatible with solubility and assay performance; otherwise, choose high-purity acetonitrile/methanol and capture waste for proper disposal.
  • Employ microscale assay formats (96/384-well) to minimize reagent consumption.

• Solvent comparison (general guidance)

  • Acetonitrile vs. methanol in LC–MS: Methanol is often considered greener (renewable sourcing, lower toxicity) but can change selectivity and backpressure; acetonitrile may give sharper peaks for hydantoins. Validate and choose based on performance vs. sustainability.
  • DMSO vs. DMF: DMSO generally preferred for lower toxicity and better EHS profile; DMF should be avoided unless necessary for solubility.

• Packaging and storage

  • Opt for single-use aliquots to limit freeze–thaw and reduce the risk of discarding partially degraded material.

Note: Isotopic labeling (13C, 2H) does not impact environmental fate meaningfully; the green focus should be on solvent choice, scale reduction, and waste management.

Pharmaceutical Uses

No excipient grade or pharmacopeial status is provided for this item. As supplied, it is designated for research use only.

• Research/manufacturing context (general)

  • Isotopically labeled mephenytoin is typically used as an analytical reference standard in pharmaceutical R&D for method development, assay calibration, and in vitro ADME investigations (e.g., CYP phenotyping, metabolic stability). It is not used as an active ingredient or excipient in finished dosage forms.
  • In bioanalytical labs, stable-isotope internal standards enable accurate quantitation of mephenytoin and metabolites in preclinical matrices to support DMPK decision-making.

• Regulatory note

  • Maintain documentation (CoA with isotopic enrichment, purity, and lot traceability) in regulated environments. Validation parameters (specificity, linearity, accuracy, precision, recovery, matrix effects, stability) should be established using the specific isotopologue supplied.

• Handling in formulation studies

  • When spiking into formulations or biological matrices for recovery studies, confirm the absence of interfering excipients and verify that the isotopic label provides clean mass separation from the analyte’s natural isotopic envelope.

No therapeutic claims are implied; this product is for laboratory research use only.

Physical Properties

Item-specific physico-chemical specifications are not provided in the Product Data and should be confirmed on the CoA/Spec Sheet for the exact isotopologue supplied.

• Item-specific values

  • Appearance: Not specified for this item; refer to CoA/Spec Sheet.
  • Melting point, boiling point, density, refractive index, UV cutoff, water content, residual solvents, isotope enrichment: Not specified for this item; refer to CoA/Spec Sheet.

• Literature/general properties for unlabeled mephenytoin (for context only; not product specifications)

  • Physical state: typically a crystalline solid (hydantoin derivative).
  • Solubility profile: sparingly soluble in water; soluble in polar aprotic organic solvents (e.g., DMSO, DMF) and moderately in alcohols; increased solubility with base due to imide acidity.
  • Acid–base behavior: cyclic imide with relatively acidic NH hydrogen (pKa of related hydantoins often ~8–10, literature), enabling partial deprotonation in basic media.
  • Lipophilicity: presence of phenyl and ethyl confers moderate hydrophobicity; unlabeled analogs exhibit measurable logP; isotope substitution does not materially change partitioning.

• Notes on isotopic labeling

  • Incorporation of 13C/2H does not significantly alter bulk thermophysical properties; minor shifts in mass and spectroscopic signatures (MS, NMR, IR) are expected and are the principal analytical differentiators.

Always verify any required numeric specification (mp, solubility, isotope enrichment) on the lot-specific CoA.

Quality and Grades

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

• Considerations specific to isotopically labeled small molecules

  • Isotopic enrichment: For quantitative LC–MS use, confirm percent enrichment and labeling positions (e.g., 13C, 2H) on the CoA. Even small amounts of unlabeled isotopologue can bias isotope-dilution assays if the analyte is abundant.
  • Chemical purity vs. isotopic purity: Both are important. Chemical purity (absence of organic impurities/byproducts) affects chromatography and ionization. Isotopic purity affects mass spectral selectivity and response factors.
  • Residual solvent and water content: These can influence accurate gravimetric preparation of standards; verify on CoA when preparing calibration stocks by weight.
  • Counter-ions/salt form: Hydantoin derivatives are commonly provided as neutral free bases; if a salt or solvate is supplied, check CoA to compute exact molarity from mass.

• What “analytical/reference standard” often implies (general)

  • Tight control of identity by MS/NMR and high chromatographic purity.
  • Traceable documentation (CoA) detailing labeling pattern, enrichment, and recommended handling.

If your application is regulated or requires traceability (e.g., quantitative bioanalysis), retain the CoA and any isotopic distribution data for method validation.

Reaction and Applications

Given its identity as an isotopically labeled mephenytoin, the compound is most relevant to analytical and biochemical applications rather than as a general-purpose synthetic reagent.

• Analytical applications (typical)

  • Stable-isotope internal standard (SIIS) for LC–MS quantification of mephenytoin and/or its metabolites (e.g., 4′-hydroxymephenytoin) in in vitro or preclinical matrices.
  • Recovery and matrix-effect correction in bioanalytical method validation (precision/accuracy, extraction recovery, ion suppression assessment).
  • Tracer in metabolic stability, microsomal/S9 incubations, and CYP phenotyping experiments to distinguish parent from background.

• Biochemical applications (literature/general)

  • Substrate probe in in vitro CYP2C19 activity assessments using human liver microsomes or recombinant enzymes; isotopic labeling facilitates tracking and quantitation of parent and metabolites by MS.

• Practical tips

  • Prepare a primary stock in dry DMSO or methanol; store single-use aliquots at −20 °C to minimize freeze–thaw and adsorption losses.
  • For enzyme assays, limit organic cosolvent to ≤1% v/v (often 0.1–0.5%) to avoid inhibiting enzyme activity; confirm solvent tolerance for your system.
  • When used as an internal standard, match retention behavior by choosing a labeling pattern that does not alter chromatography; verify no isotopic overlap with the analyte’s natural isotope cluster at the selected MS resolution.

• Not typically used for: cross-couplings, Grignard reactions, or as a general building block; see Synthetic Utility if planning method-development spiking or derivatization for analytical purposes.

Reaction Conditions

This product is primarily used in analytical and enzymology settings rather than synthetic reactions. The following are general, literature-based conditions for in vitro CYP assays and LC–MS workflows involving mephenytoin; adapt to your system and validate.

• In vitro microsomal assay (general guidance)

  • Matrix: human liver microsomes (0.1–1.0 mg/mL protein) or recombinant CYP2C19.
  • Buffer: 50–100 mM phosphate, pH 7.4, with MgCl2 (3–5 mM).
  • Substrate: mephenytoin (or isotopically labeled analog for internal standarding); final organic cosolvent ≤0.5–1% v/v (DMSO or methanol).
  • Cofactor: NADPH-regenerating system (e.g., NADP+, glucose-6-phosphate, G6PDH) or NADPH (1 mM).
  • Temperature/time: 37 °C; 5–60 min depending on enzyme source and desired turnover.
  • Quench: cold acetonitrile or methanol containing internal standard; centrifuge and analyze supernatant by LC–MS/MS.

• LC–MS/MS analysis (general)

  • Column: C18 or phenyl-hexyl; mobile phase water/acetonitrile or methanol with volatile buffer (e.g., 0.1% formic acid or ammonium acetate).
  • Detection: ESI positive mode often suitable for hydantoins; monitor parent→fragment transitions that retain the isotopic label; ensure mass separation from the unlabeled analyte’s isotope envelope.

These are non-binding guidelines. Optimize protein concentration, incubation time, and substrate levels to maintain linearity and avoid substrate depletion or enzyme inhibition.

Safety and Handling

• GHS and hazard statements

  • Signal word: Not specified for this item; refer to SDS.
  • H-statements/Pictograms/GHS classification: Not specified for this item; refer to SDS.

• General laboratory precautions (good practice for small organic imides and isotope standards)

  • Use in a fume hood to avoid inhalation of dust or solvent vapors from stock solutions.
  • Wear appropriate PPE: lab coat, safety glasses, disposable nitrile gloves. Avoid skin/eye contact and ingestion.
  • Prevent formation of dust/aerosols; handle solids gently. For solutions, cap vials promptly to minimize evaporation/contamination.

• Incompatibilities and stability (general)

  • Avoid strong bases at elevated temperature, which can promote ring opening or hydrolysis of imides.
  • Strong oxidizers and strong reducing agents may be incompatible; segregate accordingly.
  • Light and moisture can impact long-term stability of some hydantoins; store in dark, dry conditions as a precaution.

• First-aid overview (consult SDS for definitive guidance)

  • Inhalation: move to fresh air; seek medical attention if symptoms occur.
  • Skin/eye contact: rinse with water for several minutes; remove contaminated clothing; seek medical attention if irritation persists.
  • Ingestion: rinse mouth; do not induce vomiting; seek medical advice.

• Waste disposal

  • Collect solid/solution waste as organic chemical waste; dispose per institutional and local regulations.

Always defer to the product’s SDS for authoritative hazard and first-aid information.

Solvent Selection

This compound is a hydantoin derivative used most often as a reference standard or substrate in biochemical assays. Solvent choice should prioritize solubility, chemical stability, and MS-compatibility for analytical workflows.

• Practical solvent choices (general guidance)

  • DMSO: typically affords highest solubility; suitable for preparing concentrated stocks (e.g., 10–50 mM) for LC–MS or enzyme assays; dilute into aqueous buffers last to avoid precipitation.
  • DMF or NMP: alternative polar aprotic solvents when DMSO is undesirable; consider toxicity and MS background.
  • Methanol/Acetonitrile: useful for LC–MS working solutions and calibration standards; may require sonication or mild warming for dissolution.
  • Aqueous buffers: poor solubility expected at neutral pH; modest solubility increase in basic media due to imide deprotonation. Avoid prolonged high pH to prevent hydrolysis.

• Compatibility notes

  • For LC–MS, acetonitrile/water with 0.1% formic acid (or ammonium formate/acetate for negative mode) are common; verify ionization mode empirically (hydantoins often ionize in positive ESI).
  • Avoid strongly nucleophilic solvents when performing long incubations at elevated temperatures.

• Quick comparison (general)

  • Highest solubility: DMSO > DMF ≳ methanol > acetonitrile >> water (neutral pH).
  • Best for MS baselines: acetonitrile/methanol (HPLC grade) with volatile buffers.

Storage and Reconstitution

• Item-specific storage/shipping (from Product Data)

  • Storage Conditions: Store at −20 °C.
  • Shipped In: Ice chest + Ice pads.

• Stability and handling (general guidance for isotopically labeled hydantoins)

  • Protect from moisture and light. Allow vials to equilibrate to room temperature in a desiccator before opening to prevent condensation.
  • Minimize freeze–thaw by preparing single-use aliquots of stock solutions.

• Reconstitution and stock preparation (general guidance; not a specification)

  • Solvents: DMSO or methanol are commonly used to prepare primary stocks; acetonitrile may also be suitable. Sonication or gentle warming (≤40 °C) can aid dissolution.
  • Typical concentrations: 1–50 mM, depending on solubility and intended use. Filter (0.2 µm PTFE) if particulate is observed.
  • Working solutions: Dilute stocks into aqueous mobile phase or assay buffer immediately before use; keep organic cosolvent ≤1% v/v for enzyme assays.

• Solution stability (general)

  • Store frozen working stocks at −20 °C or below when compatible with solvent; avoid prolonged storage at room temperature. Verify stability as part of method validation (bench-top, autosampler, freeze–thaw).

• Research use

  • Research Use Only (as stated in Product Data). Not for human or veterinary use.

For any lot-specific details (appearance, exact solubility, isotope enrichment and placement), consult the CoA/Spec Sheet.

Structure and Identity

Brief overview: This product is an isotopically labeled form of mephenytoin intended for research use (e.g., as a stable-isotope internal standard or tracer). Specific labeling pattern (13C and/or 2H incorporation sites) is not provided in the Product Data.

• Item-specific identifiers

  • CAS: 1261398-53-1
  • Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
  • Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
  • SMILES: Not specified for this item; refer to CoA/Spec Sheet.
  • InChIKey: Not specified for this item; refer to CoA/Spec Sheet.
  • Synonym context: “Mephenytoin-C,d” denotes a carbon- and deuterium-labeled isotopologue; exact isotopic content/positions are not specified.

• Structural features (general, literature-based for unlabeled mephenytoin)

  • Core scaffold: hydantoin (imidazolidine-2,4-dione) ring.
  • Substitution: 5,5-disubstituted with phenyl and ethyl groups; N-3 methylation (3-methyl-5-ethyl-5-phenylhydantoin).
  • Functional groups: two imide carbonyls (cyclic imide), aromatic phenyl ring, aliphatic ethyl, N-methyl.
  • Stereochemistry: The 5,5-disubstitution creates a stereogenic center at C-5 in related hydantoins; mephenytoin is typically used as a racemate in many studies; enantiomeric forms (S/R) are known in literature.

• 2D description in words (literature/general)

  • A five-membered imidazolidinedione ring bearing at C-5 a phenyl and an ethyl substituent, with a methyl on the N-3 nitrogen; two carbonyls at positions 2 and 4. The isotopic label(s) (13C, 2H) are introduced at unspecified positions and do not change the connectivity.

Synthetic Utility

As an isotopically labeled analog of mephenytoin, this compound is not generally employed as a synthetic building block. Instead, it serves as an analytical standard. Nevertheless, several utility aspects may be relevant in method development and derivatization workflows.

• Functional group reactivity (general for hydantoins)

  • The cyclic imide contains acidic NH (unless fully N-alkylated); deprotonation enables formation of N-anions that can undergo alkylation or acylation under strong base. Excess base/heat may induce ring opening.
  • Aromatic ring permits electrophilic substitution on parent scaffolds; however, derivatization of an isotopic standard is uncommon and may compromise isotopic integrity.

• Analytical derivatization (general)

  • For GC–MS, hydantoins are sometimes derivatized (e.g., silylation) to improve volatility and thermal stability. If derivatization is used, confirm the isotopic label remains on the analyte fragment(s) monitored.

• Retrosynthetic value

  • Limited for this item due to its specialized labeled nature. If synthetic modifications are required (e.g., to generate isotope-labeled metabolites), start from appropriately labeled precursors and validate label placement by NMR/MS.

In most laboratories, the optimal use is as-received for LC–MS calibration, spike–recovery studies, and as a tracer in enzymology assays.

Target Specificity

Not applicable. This product is a small-molecule isotopic standard, not an antibody, enzyme inhibitor panel, or ligand with defined biological target binding data in the Product Data. For enzyme assays using mephenytoin as a substrate, consult Biological Roles and Reaction Conditions sections for general CYP2C19 assay context (literature-based).