4-Hydroxy Aceclofenac-d , CAS No.H1446525

CAS: H1446525 Cat. No.: H1446525 Formula: C₁₆H₉D₄Cl₂NO₅ Molecular Weight: 374.21

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Overview

4-Hydroxy Aceclofenac-d 4 is the deuterium labeled 4-Hydroxy Aceclofenac.

Specifications

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

This product requires cold chain shipping. Ground and other economy services are not available.

Names and Identifiers

Molecular Weight

374.21

Application Protocols

No item-specific, tested biological assay protocols are provided. As a general guide for LC–MS/MS quantitation using a deuterated internal standard:

• Spiking: Add a fixed concentration of 4-Hydroxy Aceclofenac-d to all calibration standards, QCs, and unknowns at a level comparable to the mid-calibration range of the analyte.
• Sample prep: Perform protein precipitation (3–4× volume of cold MeOH/ACN), SPE, or LLE depending on matrix cleanliness requirements. Include the internal standard prior to extraction to correct for recovery.
• Chromatography: Use a reversed-phase C18 column; mobile phase A: water + 0.1% formic acid or 5 mM ammonium acetate; mobile phase B: MeOH or ACN. Optimize gradient and temperature.
• MS detection: Monitor multiple-reaction-monitoring (MRM) transitions corresponding to the deuterated parent ion and characteristic fragments. Adjust collision energies to balance sensitivity and selectivity.
• Calibration: Use matrix-matched calibration curves; apply a 1/x or 1/x2 weighting where appropriate. Evaluate carryover and matrix effects.

These are general recommendations; finalize conditions through method development and validation per your laboratory’s SOPs. No western blot/IHC/IF/FC protocols apply to this small-molecule standard.

Biological Roles

Item-specific biological data are not provided; the following describes the role of the unlabeled scaffold in biochemical contexts (literature, non-clinical).

• Biotransformation context: 4-hydroxy aceclofenac is a Phase I oxidative metabolite formed from aceclofenac, typically via cytochrome P450–mediated aromatic hydroxylation. Hydroxylation introduces a phenolic group, increasing polarity and enabling further Phase II conjugation (e.g., glucuronidation, sulfation).
• Analytical relevance: The deuterated analog (this product) functions as an internal standard for monitoring the 4-hydroxy metabolite in in vitro systems (microsomes, hepatocytes) and in nonclinical matrices, facilitating kinetic studies (formation clearance, intrinsic clearance) and metabolite identification.
• Transport and disposition (general for arylacetic acids): carboxylate-containing molecules often interact with organic anion transporters (OATs) in hepatic/renal tissues. Phenolic hydroxylation can alter transporter affinity and plasma protein binding.
• Enzymology (general): Subsequent conjugation by UDP-glucuronosyltransferases (UGTs) or sulfotransferases (SULTs) increases aqueous solubility, supporting excretion. The regioselectivity of hydroxylation (para position) influences conjugate profiles.

Note: The above is general, literature-based biochemical context for arylacetic acid NSAID metabolites. No medical or clinical claims are made, and no item-specific biological performance is implied.

Buffer Applications

This compound is not a buffering agent. It is typically used as an analytical/internal standard rather than to control pH. For LC–MS workflows, volatile aqueous buffers (e.g., 2–10 mM ammonium acetate/formate, pH adjusted as needed) are commonly paired with organic modifiers (MeOH or ACN) to separate and detect the analyte and its deuterated analog. If preparing aqueous solutions or diluents, validate pH and ionic strength for chromatographic compatibility, but do not rely on this compound for buffering capacity.

Green Alternatives

Context: As an analytical reference standard, greener practice focuses on solvent and workflow choices rather than replacing the compound itself.

Greener solvent choices (general):

• Prefer methanol over acetonitrile when chromatographic performance is acceptable (lower toxicity, renewable sourcing potential). Consider ethanol for sample preparation when compatible with analyte stability and LC method.
• Use aqueous mobile phases with volatile, low-toxicity buffers (ammonium acetate/formate) and minimize buffer concentration to what is required for chromatographic performance.

Operational improvements:

• Miniaturize sample prep (micro-SPE, low-volume protein precipitation) to reduce solvent consumption.
• Employ UHPLC with shorter columns and smaller particle sizes to decrease run times and eluent volumes.
• Implement multi-analyte panels to consolidate analyses, further reducing solvent use per data point.

Comparison highlights (general):

• Methanol vs ACN: MeOH is greener but can lead to higher backpressure and occasionally broader peaks; ACN gives lower viscosity and sharper peaks but is less sustainable.
• Aqueous buffers vs non-volatile salts: volatile buffers are more MS-friendly and reduce hazardous waste burden compared with phosphate or sulfate buffers.

Waste handling:

• Segregate halogenated from non-halogenated organic waste; opt for MeOH/water where possible to keep waste streams non-halogenated.

Note: No item-specific stabilizers or additives are specified; consult CoA to ensure greener substitutions do not compromise analyte stability.

Pharmaceutical Uses

Therapeutic use: None. This material is supplied strictly for research use only.

Relevant roles in pharmaceutical R&D and QC (general):

• Reference standard: Used as a stable-isotope-labeled internal standard for quantifying 4-hydroxy aceclofenac in nonclinical samples, in vitro metabolism systems, or formulation/stability studies.
• Method development: Supports validation of bioanalytical methods (selectivity, linearity, accuracy, precision, recovery, matrix effects, carryover, dilution integrity) per regulatory guidance for nonclinical studies.
• Stability-indicating assays: Enables tracking of hydroxylated degradants during forced-degradation or real-time stability programs in development settings.

Pharmacopeial status: Not specified for this item; refer to CoA/Spec Sheet and relevant compendia. Isotope-labeled analogs are generally not compendial articles but may be qualified in internal standards libraries.

Manufacturing considerations: None specific—material is not intended for incorporation into drug product. Handle as a laboratory chemical with appropriate documentation (CoA, SDS), change-control, and chain-of-custody suitable for GxP environments if used in regulated labs.

Physical Properties

Item-specific properties

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
• Melting point, boiling point, density, refractive index, UV cutoff: Not specified for this item; refer to CoA/Spec Sheet.

General/literature guidance for small-molecule deuterated analytical standards of arylacetic acid scaffolds (not item-specific):

• Physical state: commonly supplied as a solid or film; may also be provided as a solution standard in acetonitrile, methanol, or DMSO.
• Solubility profile: typically soluble in polar aprotic/Protic organic solvents (MeOH, ACN, DMSO); limited water solubility in the neutral form; enhanced aqueous solubility upon basification to the carboxylate salt.
• pKa (arylacetic acids, literature): carboxylate ~3.5–4.5; phenolic OH (if present) typically ~9–10. Exact values for this item are not specified.
• logP/logD (literature, arylacetic analogs): moderate lipophilicity; deuteration has negligible effect on equilibrium partitioning but can subtly influence MS response via isotope effects.

Practical notes (general):

• For quantitative LC–MS/MS, prepare stock solutions in MeOH, ACN, or DMSO, then dilute with aqueous buffer containing 0.1% formic acid to improve chromatographic peak shape.
• Filter or centrifuge working solutions before injection to remove particulates. Validate solution stability at intended storage temperature and time.

Quality and Grades

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

Guidance on typical quality attributes for isotopically labeled small-molecule standards (general):

• Chemical purity: High purity minimizes interference in quantitative assays. Review CoA for assay (%), residual solvents, and chromatographic profile.
• Isotopic enrichment and position: For “-d” materials, the degree and location of deuteration critically affect mass shifts and isotopic interference patterns. CoA should specify D-atom count, labeling sites, and isotopic enrichment (atom % D) and state any residual protiated species.
• Identity confirmation: Expect orthogonal characterization (HRMS, 1H/13C/2H NMR where applicable, and HPLC/UPLC purity). Some suppliers also provide IR and elemental analysis.
• Residual water/peroxide/metal/UV cutoff: Not specified for this item; refer to CoA/Spec Sheet. Avoid assuming trace-spec values without documentation.
• Stabilizers: If present, stabilizers (e.g., formic acid, BHT) will be listed on the CoA; they can influence MS ionization or UV detection. None are specified here.

Fit-for-purpose considerations:

• For LC–MS/MS quantitation, ensure isotopic purity is sufficient to avoid co-elution/overlap with analyte MRM channels.
• Confirm that the salt form (free acid vs. salt) and any counterions match your method development assumptions (retention, ionization efficiency).

Reaction and Applications

Primary application domain (this item): analytical standard for quantitative and qualitative analysis of 4-hydroxy aceclofenac in biological, environmental, or formulation matrices using LC–MS/MS or LC–UV.

Use-cases (general, deuterated standards):

• Stable-isotope dilution assays (SIDA): spike a known amount of 4-Hydroxy Aceclofenac-d to compensate for matrix effects and extraction losses; calibrate using analyte-to-internal-standard response ratios.
• Method validation: assess recovery, precision, accuracy, ion suppression/enhancement, and stability in bioanalytical workflows (protein precipitation, SPE, or LLE).
• Metabolic profiling: support Phase I metabolism studies of aceclofenac by providing a co-eluting, mass-shifted internal standard for the 4-hydroxy metabolite.
• Formulation/impurity analysis: quantify hydroxylated degradants or trace impurities in stability studies.

Practical tips:

• Prepare daily working solutions freshly or verify bench-top, autosampler, and freeze–thaw stability during validation. Protect from light if phenolic chromophores are present.
• For negative ESI, use volatile ammonium buffers (e.g., 2–10 mM ammonium acetate) to enhance deprotonated ion [M–H]− signals. For positive ESI (if relevant), add 0.1% formic acid.
• Align retention times: co-inject analyte and internal standard; the isotopologue should co-elute under reversed-phase conditions, simplifying quantitation.

Note: This item is intended for research use only. No clinical or diagnostic use is implied.

Reaction Conditions

This product is not typically used as a reagent in chemical reactions. The following guidance addresses analytical handling and sample preparation conditions for LC–MS applications (general, not item-specific):

• Stock solutions: Prepare at 0.1–1.0 mg/mL in MeOH, ACN, or DMSO. Vortex until dissolved; sonication may help. Filter through 0.2 µm PTFE if particulate is present.
• Working solutions: Dilute with mobile phase or water:MeOH/ACN (50:50) containing 0.1% formic acid (positive ESI) or 2–10 mM ammonium acetate/formate (negative ESI). Keep organic ≥5% to prevent adsorption.
• Sample preparation: For plasma/serum, precipitate proteins with 3–4 volumes of cold ACN or MeOH containing the internal standard; centrifuge and transfer supernatant. Alternatively, use SPE (mixed-mode or reversed-phase) with MeOH/ACN elution.
• Chromatography: Reversed-phase C18 or phenyl-hexyl columns are common. Gradient elution from aqueous buffer to 60–95% organic in 5–10 min often resolves analyte and internal standard.
• Stability checks: Assess bench-top (2–6 h), autosampler (4–24 h at 4–10°C), freeze–thaw (3 cycles), and long-term (−20°C) stability per validation guidelines.

Note: Concentrations, times, and conditions should be optimized and validated for your matrix, instrument, and method. No reaction yields or catalytic conditions apply since this is an analytical standard.

Safety and Handling

Hazard information (item-specific)

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

General laboratory safety guidance for arylacetic acid derivatives and their deuterated standards (not item-specific):

• PPE: wear lab coat, safety glasses, and suitable gloves (e.g., nitrile). Work in a fume hood when handling powders/solutions to avoid inhalation and solvent exposure.
• Avoid generating dust and aerosols; prevent skin and eye contact. Wash thoroughly after handling.
• Incompatibilities: strong oxidizers; strong bases/acids may cause hydrolysis or salt formation; avoid prolonged exposure to elevated temperatures and light that can promote degradation.
• Analytical-use caution: deuterated standards are typically non-volatile; however, use closed containers to prevent cross-contamination of bioanalytical samples.

First-aid overview (general):

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

Additional notes:

• Always consult the product-specific SDS for authoritative hazard, toxicological, and exposure controls information.
• Waste disposal: follow institutional and local regulations for organic chemical waste; segregate halogenated solvent waste when applicable.

Solvent Selection

Applicability: As a deuterated small-molecule analytical standard, solvent choice primarily affects solubility, stability, and LC–MS performance rather than reaction outcomes.

General solvent guidance (not item-specific):

• Stock preparation: methanol (MeOH), acetonitrile (ACN), or DMSO are commonly used due to good solubility for arylacetic acid derivatives and compatibility with LC–MS.
• Working dilutions: aqueous mobile phases with 0.1% formic acid (positive mode) or 2–10 mM ammonium acetate/formate (negative mode) are typical; maintain organic content (5–60%) to control retention.
• Aqueous solubility: increases at basic pH due to carboxylate formation; however, basic conditions can promote hydrolysis in ester-containing analogs—validate stability.

Polarity/miscibility (general characteristics):

• MeOH and ACN are fully miscible with water; both support sharp chromatographic peaks. DMSO is highly polar and miscible with water but should be kept ≤1–2% in injections to avoid peak distortion.

Selection tips:

• Choose MeOH when seeking slightly stronger hydrogen-bonding interactions and lower toxicity profile; choose ACN for lower viscosity and often improved peak shape/pressure in UPLC.
• For preparative work or SPE elution, MeOH or ACN with 0.1–1% formic/acetic acid is typical.

Comparison (general):

• MeOH: greener, inexpensive, moderate elution strength.
• ACN: superior chromatographic performance in many methods; supply volatility can vary.
• DMSO: excellent solubilizer for concentrated stocks; limit in final mobile phase.

Storage and Reconstitution

Item-specific conditions

• Storage: Store at −20°C (per product data). Protect from moisture and light. Keep container tightly closed.
• Shipping: Shipped in ice chest with ice pads to maintain low temperature during transit.
• Reconstitution: Not specified for this item; refer to CoA/Spec Sheet. If supplied as a solid, prepare stock in MeOH, ACN, or DMSO. If supplied as a solution, equilibrate to room temperature before opening to prevent condensation; briefly vortex before use.

General best practices for small-molecule analytical standards:

• Aliquot stocks into low-bind, amber vials to minimize adsorption and photodegradation. Avoid repeated freeze–thaw; use single-use aliquots when possible.
• Headspace control: purge with inert gas (e.g., nitrogen) if long-term storage is planned and the compound is air/light sensitive (phenolic groups can be prone to slow oxidation).
• Stability verification: Establish solution stability at working concentration under anticipated autosampler conditions (e.g., 4–10°C over 24 h). Document long-term stability at −20°C as part of method validation.
• Compatibility: Use PTFE-lined caps; avoid reactive plastics for long-term storage of organic solutions.

Always consult the lot-specific CoA and SDS for definitive guidance on concentration, solvent compatibility, and safety prior to use.

Structure and Identity

Brief description: 4-Hydroxy Aceclofenac-d is a deuterium-labeled analog of the 4-hydroxylated aceclofenac metabolite, supplied for research and analytical use (stable-isotope standard). Specific identifiers for this catalog item are not provided below and should be confirmed against the CoA.

• 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.
• CAS: H1446525 (catalog reference; verify regulatory CAS if needed)
• Deuteration: The “-d” designation indicates incorporation of one or more deuterium atoms for use in isotope-dilution workflows (location and degree of labeling not specified; consult CoA).

Structural features (general/literature description of the unlabeled scaffold):

• Aceclofenac belongs to the arylacetic acid class related to diclofenac; the 4-hydroxy metabolite bears a phenolic OH on one aromatic ring (position assignment per name; exact substitution pattern for this labeled material should be verified from the provided structure on the CoA).
• Functional groups expected for the scaffold (literature): aryl rings, secondary anilide/amine linkage, carboxylate/ester-derived arylacetic acid moiety, and a phenolic OH at the “4-hydroxy” position.

2D structural description (literature, unlabeled core): a dichloro-substituted anilino-biphenyl/phenylacetic framework in which one ring carries a para-hydroxyl substituent; the side chain terminates in a carboxylate (or ester-derived) arylacetic acid functionality. Deuterium atoms substitute for protium at one or more defined positions (unspecified here).

Synthetic Utility

Primary value: analytical internal standard rather than a general synthetic building block. Nevertheless, deuterated analogs can offer utility in mechanistic and tracing studies.

Potential applications (general):

• Isotopic tracing: Track metabolic fate or degradation pathways of the 4-hydroxy aceclofenac scaffold via MS by following mass shifts attributable to deuterium.
• Kinetic isotope effects: Although typically subtle for aromatic/benzylic C–D bonds at ambient temperature, deuteration can be leveraged to probe hydrogen-transfer steps in oxidative pathways (research context).
• Calibration and quantitation: Serves as a co-eluting standard to correct for variable recovery and ion suppression in complex matrices.

Limitations:

• Reactivity manipulation: Deuteration is not intended to alter synthetic reactivity in a preparative sense; any effects on rate/selectivity are typically small and position-dependent.
• Structural specificity: Without item-specific labeling-site information, synthetic derivatization strategies should not assume deuterium location; obtain detailed labeling data from the CoA before planning transformations.

Takeaway: Use this compound primarily as a metrologically sound reference for LC–MS workflows and mechanistic tracing, rather than as a substrate for chemical synthesis.

Target Specificity

Not applicable. This product is a small-molecule analytical standard, not a biological macromolecule or affinity reagent. There is no antigen/epitope, clone, isotype, or species reactivity associated with this item.