Kaempferol 3-O-β-D-glucuronide - 10mM in DMSO , CAS No.22688-78-4

CAS: 22688-78-4 Cat. No.: K656701 Fórmula: C₂₁H₁₈O₁₂ Peso molecular: 462.36 Número CE: 685-695-4 PubChem CID: 5318759

GRADE & PURITY 10mM in DMSO

Storage

Protected from light,Store at -80°C

Shipped In

Dry ice packs + Cold packs

Visão geral

Kaempferol 3-O-β-D-glucuronide (Kaempferol-3-glucuronide), one conjugated kaempferol metabolite, has anti-inflammatory effect. Kaempferol 3-O-β-D-glucuronide significantly inhibits various pro-inflammatory mediators like IL-1β, NO, PGE2, and LTB4. Kaempferol 3-O-β-D-glucuronide upregulates the secretion of anti-inflammatory cytokine IL-10

IC50& Target:IL-1β|IL-10

Specifications

Especificações e pureza

10mM in DMSO

Mecanismos bioquímicos e fisiológicos

Kaempferol 3-O-β-D-glucuronide (Kaempferol-3-glucuronide), one conjugated kaempferol metabolite, has anti-inflammatory effect. Kaempferol 3-O-β-D-glucuronide significantly inhibits various pro-inflammatory mediators like IL-1β, NO, PGE2, and LTB4. Kaempfe

Condições de armazenamento de armazenamento

Protected from light,Store at -80°C

Enviado em

Dry ice packs + Cold packs

Este produto requer transporte de cadeia fria. Serviços terrestres e outros serviços econômicos não estão disponíveis.

Tipo de ação

MODULATOR

Nomes e identificadores

SMILES isoméricas

C1=CC(=CC=C1C2=C(C(=O)C3=C(C=C(C=C3O2)O)O)O[C@H]4[C@@H]([C@H]([C@@H]([C@H](O4)C(=O)O)O)O)O)O

CAS alternativo

22688-78-4

PubChem CID

5318759

Peso molecular

462.36

Application Protocols

The product is supplied for research use as a small-molecule standard. No item-specific, validated protocols are provided. Below are literature-style example workflows to aid method development (not product specifications):

• Preparation of stock solutions:

  • Weigh accurately under low light. Dissolve to 10 mM in anhydrous DMSO or to 1–5 mg/mL in water/buffer (pH 6–7). Vortex and sonicate briefly if needed.
  • Sterile filter (0.22 µm) if required for cell-free enzymatic assays.

• LC–MS/MS calibration curve (negative ESI):

  • Prepare serial dilutions (e.g., 1–1000 ng/mL) in 50:50 water:acetonitrile with 0.1% formic acid.
  • Monitor transition 461→285 m/z (neutral loss 176 Da) and, optionally, qualifier ions (e.g., 461→175 m/z) for identity confirmation (literature).

• β-Glucuronidase assay (qualitative deconjugation):

  • Incubate 10–50 µM analyte in 50 mM acetate buffer pH 5.0 with enzyme at 37°C for 1–2 h.
  • Quench with 2–3 volumes of cold acetonitrile, centrifuge, and analyze supernatant by LC–MS to verify formation of kaempferol.

• HPLC-UV monitoring:

  • Use water–acetonitrile (0.1% formic acid) gradient on C18; track at ~265 nm and ~365 nm (literature bands for flavonols).

Adjust parameters to your instrumentation and regulatory context.

Biological Roles

• General biochemical context (literature; not item-specific):

  • Kaempferol 3-O-β-D-glucuronide is a Phase II conjugate formed by UDP-glucuronosyltransferases (UGTs) from the flavonol kaempferol. Glucuronidation increases polarity and typically enhances aqueous solubility and excretion.
  • In plants, flavonol glucuronides occur as endogenous metabolites contributing to phenylpropanoid metabolism and may serve as storage/transport forms of aglycones.
  • In mammals, analogous O-glucuronides are common in xenobiotic and dietary polyphenol metabolism; they can be substrates for efflux transporters (e.g., MRPs) and targets for deconjugation by β-glucuronidase in certain tissues or microbiota.

• Analytical biomarkers:

  • Serves as a useful analytical surrogate/standard when profiling kaempferol metabolism in biological matrices (plasma, urine, tissues) using LC–MS/MS.

• Physicochemical implications:

  • The glucuronide group introduces a carboxylate, shifting distribution into aqueous compartments and altering protein-binding relative to the aglycone. Ionization state at physiological pH strongly influences transport and detection.

Note: These roles are provided for biochemical context only; no clinical or therapeutic claims are made.

Buffer Applications

This compound is not a buffering agent. However, buffer choice is critical for its analytical handling and stability.

• Practical buffer guidance (literature/general):
  • LC–MS compatibility: Use volatile buffers such as 2–10 mM ammonium formate or acetate; adjust pH 3–6 to balance chromatographic retention and minimize hydrolysis. 0.1% formic acid is common for reversed-phase separations.
  • Enzymatic assays: β-Glucuronidase reactions are often performed in acetate (pH ~5) or phosphate (pH 6–7) buffers at 37°C; verify enzyme supplier recommendations.
  • Stability considerations: Strong base accelerates glycosidic cleavage; strong acid plus heat promotes hydrolysis. Maintain moderate pH and protect from light.

If true buffering capacity is required for your workflow, select a system based on target pH and analytical compatibility rather than attempting to use this analyte as a buffer.

Green Alternatives

As a polar flavonoid glucuronide standard, solvent selection drives most of the environmental footprint. Preference should be given to aqueous systems and greener alcohols when feasible.

• Greener handling strategies:

  • Favor water or buffered aqueous media for dissolution and chromatography when compatible with detection.
  • If an organic modifier is needed, prefer ethanol over acetonitrile/methanol for preparative work, acknowledging viscosity and UV-cutoff tradeoffs.
  • Use isocratic or short-gradient LC methods to reduce solvent consumption; recycle mobile phases where validated.

• Comparison of common options (general):

  • Water–ethanol: Highest EHS profile; may reduce resolution versus ACN; higher backpressure.
  • Water–acetonitrile: Excellent chromatographic performance; higher environmental/health impact.
  • Water–methanol: Middle ground; widely available; slightly higher UV background than ACN.

• Solid-phase extraction (SPE): Employ aqueous loading and mild organic washes to minimize solvent use; elute with small volumes of aqueous methanol.

Caveat: Avoid strong acids/bases and prolonged heat to prevent hydrolysis; greener does not override stability constraints. Evaluate small-scale pilots to verify recovery and analyte stability under greener conditions.

Pharmaceutical Uses

• Formulation/excipient status: None. This compound is not used as an approved excipient.

• Research/manufacturing context (literature/general):

  • Employed as an analytical reference standard for method development, validation, and stability-indicating assays in the context of metabolism studies of dietary flavonoids.
  • Utilized in in vitro ADME workflows (e.g., UGT metabolism, transporter interactions) to calibrate and qualify bioanalytical assays (LC–MS/MS, HPLC-UV).

• Pharmacopeial status: No monograph known for this specific glucuronide. Any specifications must be defined in an in-house method or research protocol.

No therapeutic or clinical claims are associated with this material; it is supplied strictly for research use only.

Physical Properties

• Item-specific specifications (this lot):

  • Appearance: Not specified for this item; refer to CoA/Spec Sheet.
  • Grade/Purity: Not specified for this item; refer to CoA/Spec Sheet.

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

  • Physical state: typically a yellow to off-yellow amorphous to microcrystalline solid for flavonol glucuronides.
  • Molecular weight: ~462.36 g/mol (see Structure & Identity; literature).
  • Ionization: phenolic and carboxylic functionalities; expected to exist partly as carboxylate at neutral pH, enhancing aqueous solubility.
  • Solubility profile (literature/typical):
    • Water/buffer: moderate to high solubility versus the aglycone due to glucuronide; solubility improved at pH > 6 from carboxylate formation.
    • Organic: soluble in DMSO; variably soluble in methanol and ethanol; poor in nonpolar solvents (e.g., hexanes, toluene).
  • UV-Vis (literature for kaempferol conjugates): strong absorption in Band II (~260–270 nm) and Band I (~360–370 nm), useful for HPLC-UV tracking; exact λmax depends on solvent/pH.
  • LogP/logD (prediction/literature): low logP; logD (pH 7.4) expected negative due to ionization of the glucuronide carboxyl.
  • Melting/decomposition: flavonoid glucuronides commonly show broad decomposition rather than a sharp MP; specific MP not established for this item.

Notes: Provide analytical confirmation (HPLC, HRMS, NMR) against your method; do not use these literature values as product specifications.

Quality and Grades

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

• Research-use designation:

  • This product is labeled For research use only. It is not intended for human or veterinary use, diagnostic procedures, or as a food additive.

• Interpreting common grades (general guidance):

  • Analytical/reference standard grade: Prioritized for identity and purity suitable for chromatography (HPLC/LC–MS) and quantitation; typically supported by NMR, MS, and HPLC purity on CoA.
  • HPLC grade solvents/reagents: Low UV background and low nonvolatile residue, appropriate when preparing calibration curves for UV or MS detection.

• Stabilizers/additives: None specified for this item; if stabilizers are present, they will be listed on the CoA/Spec Sheet.

• Quality verification recommendations (practical tips):

  • Confirm identity by LC–MS (negative ESI is typically sensitive for glucuronides) and by diagnostic MS/MS neutral loss of 176 Da (literature) to the aglycone (m/z 285 for kaempferol [M–H]−).
  • Verify purity by HPLC-UV at ~260–270 and ~360–370 nm (literature absorbance bands for flavonols) and by 1H/13C NMR in DMSO-d6 or methanol-d4.
  • Where trace moisture or buffer content matters, conduct Karl Fischer on arrival; hydrate state can subtly affect weighing and solubility for sugar conjugates.

Reaction and Applications

This compound is primarily used as an analytical reference standard and biochemical probe rather than as a stoichiometric reagent.

• Analytical/biochemical applications (literature):

  • LC–MS/MS metabolite identification and quantification as a Phase II conjugate of kaempferol. Diagnostic neutral loss of 176 Da (glucuronide) from [M–H]− 461 → 285 for MS/MS transitions in negative ESI.
  • Enzymology: Substrate or standard in UDP-glucuronosyltransferase (UGT) activity assays and in β-glucuronidase-mediated deconjugation studies.
  • Transport studies: Probe for MRP/OATP-mediated transport of glucuronides in membrane vesicle or cell systems (method development/reference only).

• Chemical reactivity (general guidance):

  • Acid/base sensitivity: Strong acid at elevated temperature can hydrolyze the glycosidic bond to release kaempferol. Strong base can induce degradation and cleavage.
  • Oxidation: Phenolic oxidation possible; minimize exposure to oxidants and air/metal catalysts; add antioxidants only if compatible with analysis.

• Practical tips:

  • For MS work, minimize salts and use volatile buffers (ammonium acetate/formate). Maintain low light and temperature during sample prep to preserve integrity.
  • For enzymatic hydrolysis controls, include parallel incubations without enzyme and with heat-inactivated enzyme to assess non-enzymatic loss.

Note: No medical/clinical applications are implied; this information supports laboratory method development and chemical handling.

Reaction Conditions

The compound is most often used in analytical and enzymatic workflows. The following are literature/general conditions; validate and optimize for your system.

• Enzymatic deglucuronidation (β-glucuronidase):

  • Buffer: 50–100 mM sodium acetate, pH 5.0–5.5 (or phosphate pH 6–7 for certain enzymes).
  • Temperature: 37°C.
  • Time: 0.5–4 h depending on enzyme units and matrix.
  • Monitoring: LC–MS/MS for disappearance of [M–H]− 461 and appearance of kaempferol [M–H]− 285 (neutral loss 176 Da).

• Acidic hydrolysis (chemical):

  • Reagent: 0.1–1 M HCl or H2SO4 in aqueous methanol/water.
  • Temperature: 60–90°C.
  • Time: 0.5–2 h; minimize exposure to prevent aglycone degradation.
  • Workup: Neutralize, extract aglycone into ethyl acetate, dry, and analyze.

• LC–MS analytical method (example conditions):

  • Column: C18, 2.1 × 100 mm, 1.7–3 µm.
  • Mobile phase A/B: Water + 0.1% formic acid / Acetonitrile + 0.1% formic acid.
  • Gradient: 5% B to 40% B over 10–15 min.
  • Detection: ESI(−), monitor 461→285 m/z; UV 260–270 and 360–370 nm as secondary detection.

All parameters above are illustrative literature-style guidance and not product specifications.

Safety and Handling

• Item-specific hazard information (GHS):

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

• General laboratory safety guidance (literature/practice; not a substitute for SDS):

  • Expected hazards: Polyphenolic glycosides are generally of low acute toxicity but may cause irritation to eyes/skin/respiratory tract. Handle as a chemical of unknown toxicity.
  • PPE: lab coat, safety glasses, and appropriate chemical-resistant gloves (e.g., nitrile). Use in a fume hood to minimize dust/aerosol exposure.
  • Avoid: inhalation of powders, contact with skin/eyes. Prevent moisture ingress during weighing to avoid clumping and ensure accurate massing.
  • Incompatibilities: Strong oxidizers may degrade phenolic systems; strong bases or strong acids can cleave the glycosidic bond and/or induce degradation. Protect from prolonged exposure to light (photo-labile conjugated system) and heat.
  • First aid (overview):
    • Inhalation: move to fresh air; seek medical advice if symptoms persist.
    • Skin: wash with soap and water; remove contaminated clothing.
    • Eyes: rinse cautiously with water for several minutes; remove contact lenses; seek medical attention if irritation continues.
    • Ingestion: rinse mouth; do not induce vomiting; seek medical attention.
  • Fire safety: Organic solid; use CO2, dry chemical, or foam. Combustion may produce CO/CO2 and phenolic decomposition products.

Always consult the product-specific SDS for authoritative hazard, exposure limits, and response procedures.

Solvent Selection

• Polarity/ionization profile (general):

  • Polar, polyhydroxylated phenolic glucuronide with a carboxylic acid; predominantly deprotonated at neutral pH, favoring aqueous solubility.

• Recommended solvents (use-case dependent):

  • Stock preparation: DMSO (5–20 mg/mL typical), then dilute into aqueous buffer; or dissolve directly in water/aqueous buffer when pH ≥ 6 to aid ionization of the glucuronide.
  • Chromatography: Water–acetonitrile or water–methanol with 0.1% formic acid (or 5–10 mM ammonium formate/acetate) for LC–MS. For isolation, consider buffered aqueous methanol to minimize hydrolysis.

• Miscibility/compatibility (literature/typical):

  • Miscible with polar protic solvents (water, methanol, ethanol). Poorly soluble in nonpolar solvents (hexanes, toluene, MTBE).
  • Avoid strong basic media (can accelerate β-elimination or glycosidic cleavage) and strong acids at elevated temperature (acidic hydrolysis).

• When to choose vs alternatives:

  • Compared with aglycone kaempferol, the glucuronide is more water-compatible and better suited for bioanalytical LC–MS in aqueous mobile phases.
  • For NMR, methanol-d4 or DMSO-d6 provide sharper resonances than D2O due to exchange broadening of phenolics; add a small amount of base (e.g., NaOD) only if needed for solubility, balancing risk of hydrolysis.

Storage and Reconstitution

• Item-specific storage and shipping (from Product Data):

  • Storage Conditions: Protected from light, Store at -80°C.
  • Shipped In: Dry ice packs + Cold packs.

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

• Stability guidance (general):

  • Minimize exposure to light, moisture, and elevated temperature. Flavonoid glucuronides can undergo hydrolysis and oxidative degradation.
  • For working stocks, prepare small aliquots to avoid repeated freeze–thaw cycles. Store aliquots at ≤ −20°C short-term (days to weeks) or −80°C long-term, protected from light.

• Reconstitution recommendations (general; not a specification):

  • DMSO stock: Dissolve to 5–20 mg/mL. Mix gently; brief sonication can help. Store aliquots at −20 to −80°C.
  • Aqueous stock: Dissolve in water or buffer (pH 6–7) to 0.1–5 mg/mL; filter if needed. Use promptly or freeze in aliquots at −80°C.
  • Avoid strong acids/bases and metal contaminants during preparation; use high-purity solvents (HPLC or LC–MS grade) for analytical applications.

• Shelf-life: Not specified for this item; refer to CoA/Spec Sheet.

Always refer to the product label and CoA/SDS for definitive storage and handling instructions.

Structure and Identity

Kaempferol 3-O-β-D-glucuronide is a flavonol (kaempferol) conjugated at the 3-hydroxyl with a β-D-glucuronic acid moiety (O-glycosidic linkage), yielding a highly oxygenated polyphenolic glycoside.

• Item-specific identifiers from Product Data:

  • CAS: 22688-78-4
  • CID (PubChem): 5318759
  • InChIKey: Not specified for this item; refer to CoA/Spec Sheet.
  • SMILES: Not specified for this item; refer to CoA/Spec Sheet.

• Literature/computed identifiers and features (for reference only; not item specifications):

  • Molecular formula (literature): C21H18O12
  • Molecular weight (literature): ~462.36 g/mol (kaempferol 286.24 + glucuronyl 176.03)
  • Core scaffold: flavonol backbone (3-hydroxyflavone), rings A (resorcinol-type), B (para-hydroxylated phenyl), and C (chromen-4-one).
  • Conjugation site: O-glycosidic bond at the 3-hydroxyl of kaempferol; β-anomeric configuration of D-glucuronic acid.
  • Functional groups: phenolic OH (multiple), conjugated ketone (4-one), ether glycosidic linkage, carboxylate (glucuronic acid, typically present as free acid under neutral conditions), multiple aromatic rings.

• 2D structural description (verbal): A tricyclic chromenone (flavonol) core bearing phenolic OH groups at 3 (substituted by the glycoside), 5, and 7 positions on the A/C rings, with a para-hydroxylated phenyl (B ring) at C2. The 3-position bears a β-D-glucuronyl residue attached via an O-linkage; the sugar ring terminates in a carboxylic acid at C6 of the hexose (uronic acid).

Synthetic Utility

Although primarily an analytical standard, Kaempferol 3-O-β-D-glucuronide offers utility as a chemical probe and transformation substrate.

• Transformations (literature/general):

  • Enzymatic hydrolysis: β-Glucuronidase cleaves the 3-O-glycosidic bond to regenerate kaempferol (aglycone) for comparative analyses.
  • Chemical hydrolysis: Acid-catalyzed cleavage (aqueous mineral acids, heat) affords the aglycone; conditions must be carefully controlled to avoid degradation of the flavonol core.
  • Derivatization: Phase transfer or carbodiimide-mediated coupling at the glucuronide carboxyl can provide amide/ester derivatives for tracer or immobilization applications.

• Retrosynthetic note:

  • Construction typically arises from glycosylation of protected kaempferol at the 3-OH with a suitably activated glucuronic acid donor (e.g., trichloroacetimidate donor) followed by global deprotection and ester hydrolysis to the free acid.

• Utility in methods development:

  • Serves as a matrix-relevant standard for validating recovery in SPE workflows targeting polar conjugates.
  • Useful for optimizing reversed-phase vs HILIC separations of acidic polyphenol conjugates.

These uses are methodological and synthetic in nature and do not imply biological efficacy.

Target Specificity

Not applicable. This product is a small-molecule flavonoid glucuronide, not a biological targeting reagent or antibody. No antigen/epitope specificity, clone, isotype, or species reactivity is associated with this item.