3'-Hydroxy Warfarin - ≥95% , CAS No.30992-81-5

CAS: 30992-81-5 Cat. No.: H974681 Fórmula: C₁₉H₁₆O₅ Peso molecular: 324.33 PubChem CID: 54691382

GRAU & PUREZA ≥95%

Storage

Room temperature

Especificações

Especificações e pureza

≥95%

Condições de armazenamento de armazenamento

Room temperature

Pureza

≥95%

Nomes e identificadores

Sorrisos canónicos

CC(=O)CC(C1=CC(=CC=C1)O)C2=C(C3=CC=CC=C3OC2=O)O

IUPAC Name

4-hydroxy-3-[1-(3-hydroxyphenyl)-3-oxobutyl]chromen-2-one

InChIKey

DDKYQGKCKYAWAT-UHFFFAOYSA-N

INCHI

1S/C19H16O5/c1-11(20)9-15(12-5-4-6-13(21)10-12)17-18(22)14-7-2-3-8-16(14)24-19(17)23/h2-8,10,15,21-22H,9H2,1H3

SMILES isoméricas

CC(=O)CC(C1=CC(=CC=C1)O)C2=C(C3=CC=CC=C3OC2=O)O

PubChem CID

54691382

Peso molecular

324.33

Taxonomic Classification

Taxonomy Tree

• Kingdom Organic compounds
  • Superclass Phenylpropanoids and polyketides
    • Classe Coumarins and derivatives
      • Subclass Hydroxycoumarins

• Kingdom: Organic compounds
• Superclass: Phenylpropanoids and polyketides
• Classe: Coumarins and derivatives
• Subclass: Hydroxycoumarins
• Intermediate Tree Nodes: Not available
• Direct Parent: 4-hydroxycoumarins
• Alternative Parents: 1-benzopyrans Pyranones and derivatives 1-hydroxy-4-unsubstituted benzenoids 1-hydroxy-2-unsubstituted benzenoids Benzene and substituted derivatives Vinylogous acids Heteroaromatic compounds Lactones Ketones Oxacyclic compounds Organic oxides Hydrocarbon derivatives
• Molecular Framework: Aromatic heteropolycyclic compounds
• Substituents: 4-hydroxycoumarin - Benzopyran - 1-benzopyran - 1-hydroxy-4-unsubstituted benzenoid - 1-hydroxy-2-unsubstituted benzenoid - Phenol - Pyranone - Monocyclic benzene moiety - Pyran - Benzenoid - Heteroaromatic compound - Vinylogous acid - Lactone - Ketone - Oxacycle - Organoheterocyclic compound - Organic oxygen compound - Organooxygen compound - Carbonyl group - Hydrocarbon derivative - Organic oxide - Aromatic heteropolycyclic compound
• Descrição: This compound belongs to the class of organic compounds known as 4-hydroxycoumarins. These are coumarins that contain one or more hydroxyl groups attached to C4-position the coumarin skeleton.
• External Descriptors: Not available

Data sources

1. Djoumbou Feunang Y, Eisner R, Knox C, Chepelev L, Hastings J, Owen G, Fahy E, Steinbeck C, Subramanian S, Bolton E, Greiner R, and Wishart DS. ClassyFire: Automated Chemical Classification With A Comprehensive, Computable Taxonomy. Journal of Cheminformatics, 2016, 8:61.

DOI: 10.1186/s13321-016-0174-y

Propriedades químicas e físicas

• Peso molecular: 324.300 g/mol
• XLogP3: 2.300
• Hydrogen Bond Donor Count: 2
• Hydrogen Bond Acceptor Count: 5
• Rotatable Bond Count: 4
• Exact Mass: 324.1 Da
• Monoisotopic Mass: 324.1 Da
• Topological Polar Surface Area: 83.800 Ų
• Heavy Atom Count: 24
• Formal Charge: 0
• Complexity: 538.000
• Isotope Atom Count: 0
• Defined Atom Stereocenter Count: 0
• Undefined Atom Stereocenter Count: 1
• Defined Bond Stereocenter Count: 0
• Undefined Bond Stereocenter Count: 0
• The total count of all stereochemical bonds: 0
• Covalently-Bonded Unit Count: 1

Application Protocols

No vendor-validated protocols are specified for this item; the following are general, literature-based examples for research use.

• LC–MS/MS calibration standards (typical workflow):

  • Prepare a 10 mM stock in DMSO (or MeOH) under low light. Serially dilute with 50:50 MeOH:water (0.1% formic acid) to prepare working standards (e.g., 0.5–1000 ng/mL range). Store aliquots at −20 °C protected from light; avoid repeated freeze–thaw.
  • Matrix-matched QCs: spike blank plasma or incubation matrix to desired levels (e.g., LLOQ, low, mid, high). Process alongside samples (protein precipitation with ACN or MeOH; or SPE as validated).

• In vitro enzyme assay (CYP phenotyping example):

  • Incubate racemic warfarin (5 µM) with recombinant CYP isoforms or human liver microsomes in phosphate buffer (pH 7.4) at 37 °C with NADPH regenerating system for 20–30 min.
  • Quench with 2–3 volumes of ice-cold ACN containing internal standard. Centrifuge and analyze supernatant by LC–MS/MS using transitions selective for 3'-hydroxywarfarin.

• Stability check:

  • Assess bench-top, autosampler, freeze–thaw, and long-term stability according to bioanalytical method validation guidance to ensure accurate quantitation.

These examples are informational; optimize for your instruments and regulatory framework.

Biological Roles

3'-Hydroxy Warfarin is a known phase I metabolite in the biotransformation of warfarin.

• Metabolic context (literature/general):

  • Formed via aromatic ring hydroxylation of the warfarin phenyl group at the 3'-position by hepatic cytochrome P450 enzymes (reports implicate CYP1A2, CYP2C9, and CYP3A isoforms depending on species and conditions).
  • Undergoes subsequent phase II conjugation (e.g., O-glucuronidation by UGTs; O-sulfation), increasing polarity for excretion.

• Biochemical properties:

  • Additional phenolic OH increases hydrogen-bonding capacity and decreases lipophilicity relative to the parent compound.
  • Retains the 4-hydroxycoumarin pharmacophore; literature describes variable residual activity at vitamin K cycle enzymes for different hydroxywarfarin isomers. Specific potency and protein-binding parameters are assay-dependent.

• Research significance:

  • Serves as a marker for specific CYP-isoform contributions in phenotyping panels.
  • Useful in mechanistic studies of stereoselective metabolism (R/S warfarin biotransformation) and inter-individual variability in oxidative pathways.

No claims are made regarding clinical efficacy or safety; this material is intended strictly for research use.

Buffer Applications

This compound is not used as a buffering agent. It does not constitute a conventional acid–base buffer system.

• Practical handling in aqueous buffers (literature/general):
  • Prepare concentrated stocks in DMSO or ethanol, then dilute into assay buffers (e.g., phosphate, Tris) to the desired final concentration, keeping cosolvent low (≤0.1–1% v/v as assay permits).
  • For solubility-limited applications, mildly basic buffers (pH > 7.5) can increase solubility via phenolate formation; verify assay compatibility.

For experiments requiring defined pH control, select an appropriate biological buffer (phosphate, HEPES, MOPS, etc.) independent of this analyte.

Green Alternatives

While the compound itself is fixed, greener choices can be made in its handling and analytical workflows.

• Solvent selection (literature/general):

  • Prefer ethanol or methanol over chlorinated solvents for sample preparation where feasible.
  • For extraction/sample cleanup, consider ethyl acetate or methyl tert-butyl ether (MTBE) rather than dichloromethane (DCM), balancing recovery and matrix effects.
  • For chromatographic mobile phases, use water–acetonitrile or water–methanol; avoid halogenated modifiers.

• Small comparison (general):

  • ACN vs MeOH: ACN often gives sharper peaks and lower backpressure; MeOH is less toxic and more sustainable but may alter selectivity and raise pressure.
  • DMSO vs DMF for stocks: DMSO has a better EHS profile than DMF; both solubilize phenolic coumarins well.

• Operational improvements:

  • Minimize solvent volumes via micro-extractions and 96-well platforms.
  • Employ UHPLC to reduce solvent consumption and analysis time.
  • Use amber glassware and protect from light to reduce need for re-preparation (waste prevention).

Note: Ensure any solvent switch maintains analyte recovery, stability, and compatibility with detection (validate by recovery and precision studies).

Pharmaceutical Uses

No excipient or formulation role is specified for this item; it is supplied for research use only.

• Typical research/manufacturing-related uses (literature/general):

  • Analytical reference standard for method development, system suitability, and calibration in bioanalysis of warfarin and its metabolites.
  • Internal/secondary standard candidate for in vitro ADME studies (enzyme kinetics, microsomal turnover) after appropriate qualification.
  • Impurity/metabolite identification aid in process and metabolite profiling.

• Pharmacopeia/compendial status: Not specified for this item; refer to CoA/Spec Sheet. If compendial use is required, verify against specific monograph requirements or validated reference standards.

No therapeutic or clinical claims are made for this product.

Physical Properties

• Item-specific data from Product Data:

  • Appearance: Not specified for this item; refer to CoA/Spec Sheet.
  • Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
  • Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.

• Literature/general properties (for reference; not item specifications):

  • Approx. molecular weight: ~324.33 g/mol (C19H16O5, literature for 3'-hydroxywarfarin).
  • Acid-base behavior: possesses two phenolic OH groups; the 4-hydroxycoumarin system exhibits tautomerism and an acidic phenolic proton (warfarin pKa ~4.9–5.2; hydroxylation on the phenyl ring may slightly modulate acidity; literature values vary by solvent/ionic strength).
  • LogP/logD: warfarin logP ~3.0; additional phenolic OH generally reduces lipophilicity (3'-hydroxy analog expected slightly lower; consult specific database values).
  • Solubility profile: sparingly soluble in water at neutral pH; increased solubility in basic aqueous media due to phenolate formation; readily soluble in polar organic solvents (DMSO, DMF, methanol, ethanol, acetone; literature).
  • UV–Vis: coumarin chromophore gives strong absorption in near-UV; additional phenol may slightly red-shift/alter intensity (literature). Exact λmax not specified for this item.
  • Solid-state form: typically an off-white to light tan solid for related phenolic coumarins; actual appearance not specified for this item.

Note: For authoritative, item-specific values (mp, purity, water/peroxide/metal limits, refractive index, UV cutoff), refer to the Certificate of Analysis (CoA)/Spec Sheet.

Quality and Grades

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

• Interpreting common grades for this compound class (general guidance):

  • Analytical/reference standard grade: optimized for identity/purity traceability (often accompanied by CoA with NMR/LC-MS purity). Appropriate for bioanalytical assays (e.g., LC–MS/MS calibration of warfarin metabolite panels).
  • Research grade: suitable for discovery chemistry, enzyme assays, and method development where ultra-trace impurity control is less critical than for certified standards.

• Stabilizers: Not specified for this item; refer to CoA/Spec Sheet. Phenolic coumarins typically do not require stabilizers when stored dry and protected from light; if present, stabilizers should be disclosed on the CoA.

• What to check on receipt:

  • Purity by HPLC/LC–MS and water content (if applicable) on the accompanying CoA.
  • Identity confirmation (1H/13C NMR consistent with 3'-phenyl hydroxylation; MS [M+H]+ ~325, literature).
  • Batch-specific storage/shipping notes (temperature, light protection) and any declared residual solvents.

Reaction and Applications

This compound is primarily used as a reference/analytical standard and biochemical probe rather than as a synthetic reagent. Nonetheless, it is relevant to several workflows:

• Bioanalytical/PK workflows (literature/general):

  • Calibration/quality controls for LC–MS/MS quantification of warfarin metabolites in microsomal, hepatocyte, or plasma matrices.
  • Method development for separation of positional hydroxywarfarin isomers (2'-, 3'-, 4'-, 6-, 7-, 8-hydroxywarfarins) using reversed-phase or chiral stationary phases.

• Enzymology applications:

  • Substrate/product standards for CYP450 phenotyping (notably CYP2C9, CYP1A2, CYP3A isoforms implicated in aromatic hydroxylation of warfarin; literature).
  • Kinetic studies of metabolite formation/clearance, glucuronidation (UGT enzymes), and sulfation.

• Chemical reactivity context (general):

  • The phenolic OH groups can undergo O-acylation/O-alkylation; the benzylic ketone can participate in typical carbonyl chemistry (e.g., oxime/hydrazone formation) for derivatization in analytical method development.

• Practical tips:

  • Prepare fresh working solutions; phenols can slowly oxidize—minimize air/light exposure.
  • Validate isomer identity via orthogonal methods (MS/MS fragment ions, diagnostic NMR shifts for phenolic substitution pattern).

Reaction Conditions

Representative literature conditions involving 3'-hydroxywarfarin focus on biotransformations and analytical method development rather than bulk synthesis. The following are general, literature-based guidelines (not item-specific specifications):

• In vitro microsomal incubations (formation from warfarin):

  • Matrix: human liver microsomes (0.2–1.0 mg/mL protein) in phosphate buffer (pH 7.4) with MgCl2 (3–5 mM).
  • Cofactors: NADPH-regenerating system (e.g., NADP+, glucose-6-phosphate, G6PD).
  • Substrate: warfarin (1–20 µM), 37 °C, 10–60 min; monitor 3'-hydroxy isomer formation by LC–MS/MS.

• UGT-mediated conjugation (of 3'-hydroxywarfarin):

  • Matrix: human liver microsomes or recombinant UGTs, pH 7.4–7.8 Tris buffer.
  • Cofactor: UDPGA (2–5 mM), Mg2+ (5–10 mM); 37 °C, 15–60 min.

• LC–MS/MS method hints:

  • Mobile phase: water (0.1% formic acid or ammonium acetate) with ACN or MeOH gradient.
  • Column: C18, 2.1 × 50–100 mm, 1.7–3 µm; 30–45 °C.
  • Detection: negative or positive ESI; monitor [M–H]− ~323 or [M+H]+ ~325 (literature), with characteristic coumarin fragments.

• Chemical derivatization (for detection):

  • Phenolic acylation with anhydrides/acid chlorides (pyridine/DMAP) at 0–25 °C to form more hydrophobic derivatives for GC or altered LC selectivity.

Always validate conditions for your system; yields, rates, and selectivity depend on enzyme source, matrix, and analytical configuration.

Safety and Handling

• GHS/SDS information for this item:

  • Signal word, hazard statements, pictograms, and classification: Not specified for this item; refer to the SDS.

• General laboratory precautions (literature/general guidance):

  • Avoid inhalation of dusts and contact with skin/eyes. Handle in a chemical fume hood.
  • Personal protective equipment: lab coat, safety glasses, and appropriate chemical-resistant gloves.
  • Coumarin derivatives/phenolic compounds may cause irritation; some warfarin metabolites are bioactive. Practice strict hygiene and minimize exposure.
  • Do not ingest. For research use only.

• Incompatibilities and stability (general):

  • Avoid strong oxidizers and strong bases/acids for prolonged periods; phenolic and lactone functionalities can undergo hydrolysis or oxidative degradation.
  • Protect solutions from light to prevent photodegradation of the coumarin chromophore; use amber vials where possible.

• First-aid overview (consult SDS for full instructions):

  • Inhalation: move to fresh air; seek medical attention if symptoms persist.
  • Skin contact: wash with soap and water; remove contaminated clothing.
  • Eye contact: rinse cautiously with water for several minutes; remove contact lenses if present and easy to do; seek medical advice if irritation continues.
  • Ingestion: rinse mouth; do not induce vomiting; seek medical attention.

• Spill and waste: Collect solids with minimal dust generation. Dispose in accordance with local regulations and institutional policies.

Solvent Selection

3'-Hydroxy Warfarin is an aromatic, moderately polar, phenolic lactone with limited aqueous solubility at neutral pH and good solubility in polar organics.

• Practical solvent choices (literature/general):

  • Strongly recommended for stocks: DMSO (high solubility, inert under short-term use), DMF.
  • Also suitable: methanol, ethanol, acetone, acetonitrile (moderate to good solubility); warm gently if needed.
  • Aqueous media: sparing at neutral pH; solubility increases in basic buffers (phenolate formation). If biological compatibility permits, prepare a concentrated DMSO stock and dilute into buffer, keeping final DMSO ≤0.1–1% v/v as assay allows.

• Polarity/dielectric context (literature):

  • LogP for parent warfarin ~3; addition of a phenolic OH reduces lipophilicity modestly, aiding miscibility with polar organics while remaining largely hydrophobic in water.

• When to choose vs alternatives:

  • For LC–MS standards: use acetonitrile or methanol with 0.1% formic acid or ammonium acetate for improved peak shape; DMSO for high-concentration master stocks.
  • For enzyme assays: DMSO stock diluted into phosphate or Tris buffers; consider minimal organic cosolvent to avoid enzyme inhibition.

• Practical note: Filter stock solutions (0.2 µm PTFE) for particulate removal before analytical injections. Use amber glass vials to minimize photolysis.

Storage and Reconstitution

• Item-specific storage from Product Data:

  • Storage Conditions: Room temperature.
  • Shipped in: Not specified for this item; refer to CoA/Spec Sheet.

• General storage guidance (literature/general):

  • Store dry, tightly closed, and protected from light (amber container) to minimize photodegradation and oxidation of phenolic functionalities.
  • For long-term storage of solutions, prefer aliquoting and keeping at ≤−20 °C in amber vials; minimize headspace to reduce oxidative degradation. Avoid repeated freeze–thaw.

• Reconstitution recommendations (general):

  • Prepare concentrated stock solutions in DMSO (e.g., 10–50 mM) or methanol/ethanol. Mix gently until fully dissolved; brief sonication may assist.
  • For aqueous use, dilute the organic stock into buffer with vigorous mixing to avoid precipitation; maintain final organic content appropriate for the assay.
  • Verify clarity; if particulates persist, pass through a 0.2 µm PTFE syringe filter.

• Stability notes:

  • Working solutions are best prepared fresh daily or verified by stability studies (chromatographic purity/response over time). If stability-indicating methods detect degradation, discard and re-prepare.

Always refer to the item’s CoA/SDS for definitive storage, handling, and stability information.

Structure and Identity

3'-Hydroxy Warfarin is a hydroxylated metabolite of the 4-hydroxycoumarin anticoagulant warfarin, bearing an additional phenolic OH on the pendant phenyl ring at the 3'-position.

• Item-specific identifiers (from Product Data):

  • CAS: 30992-81-5
  • SKU: H974681
  • InChIKey: Not specified for this item; refer to CoA/Spec Sheet. (Catalog lists "60576", which is not in standard InChIKey format.)
  • SMILES: Not specified for this item; refer to CoA/Spec Sheet.

• Computed/literature identity (for reference; not item-specific specifications):

  • Common name: 3'-Hydroxywarfarin; 3-Hydroxywarfarin (meta-hydroxylated phenyl)
  • Molecular formula (literature): C19H16O5
  • Molecular weight (literature): ~324.33 g/mol
  • Representative SMILES (literature): O=c1oc2cc(O)ccc2c(=O)c1CC@Hc3cccc(c3)O
    • Note: warfarin has a single stereogenic center; materials are commonly racemic unless specified.
  • InChIKey (literature): representative keys include 27-character strings; consult a structure database for the exact isomer.

• Structural features (descriptive):

  • 4-hydroxycoumarin (chromen-2-one) core featuring a 4-phenolic OH and a lactone carbonyl.
  • Side chain at C-3: 3-oxo-1-phenylbutyl substituent with one stereocenter at the benzylic position.
  • Pendant phenyl ring substituted with an additional phenolic OH at the 3' position (meta to the side-chain attachment).
  • Functional groups: phenols (2×), conjugated lactone (coumarin), aromatic rings, benzylic ketone.

Synthetic Utility

While primarily a metabolite standard, 3'-Hydroxy Warfarin contains functional handles that permit derivatization useful in analytical chemistry and probe development.

• Functional group reactivity (general):

  • Phenolic OH groups: O-acylation, O-alkylation, carbamate/carbonate formation for pro-masks or tagging; selective protection strategies can distinguish the more acidic coumarin OH from the phenyl OH under carefully chosen conditions.
  • Benzylic ketone: formation of oximes/hydrazones for stabilization/labeling; potential reduction to secondary alcohols; enolization under basic conditions for isotope exchange studies.
  • Aromatic rings: electrophilic substitution limited by deactivation from phenolic groups; cross-coupling requires prior halogenation (not inherent to this structure).

• Applications in synthesis/analysis:

  • Derivatization to increase molar absorptivity/ionization efficiency for chromatographic detection.
  • Immobilization via linker chemistry (e.g., carbonate formation) for affinity or metabolism-on-a-chip studies.

• Retrosynthetic note:

  • Accessed from warfarin by regioselective aromatic hydroxylation (chemical or biocatalytic), or by constructing the coumarin core followed by side-chain installation and directed hydroxylation on the aryl ring (literature strategies).

Target Specificity

Not applicable to this product category. 3'-Hydroxy Warfarin is a small-molecule metabolite standard, not an antibody, enzyme, or affinity reagent.

• If used in biochemical studies, it may interact with proteins involved in the vitamin K cycle or drug-metabolizing enzymes; however, no item-specific target data are provided. Any such interactions should be characterized within the context of the specific assay.