Nicotine N-β-D-glucuronide-d , CAS No.329002-74-6

CAS: 329002-74-6 Cat. No.: N1444242 Formula: C₁₆H₁₉D₃N₂O₆ Peso molecolare: 341.37

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Panoramica

Nicotine N-β-D-glucuronide-d 3 (Nicotine N-glucuronide-d 3 ) is a deuterium labeled compound.

Specifications

Condizioni di conservazione di stoccaggio

Store at -20°C

Spedito in

Ice chest + Ice pads

Questo prodotto richiede spedizione a catena fredda. I servizi di terra e altri servizi economici non sono disponibili.

Nomi e identificatori

Peso molecolare

341.37

Application Protocols

No item-specific validated protocols are provided for this product. The following general outline reflects common LC–MS/MS usage for deuterated small-molecule standards (informational only):

• Stock preparation: Dissolve an accurately weighed amount in water or 50:50 water:methanol to a convenient concentration (e.g., 0.1–1.0 mg/mL). Exact solubility and concentration for this item are not specified; consult CoA/Spec Sheet.
• Internal standard working solution: Dilute stock into mobile phase or extraction solvent to yield a target on-column amount comparable to native analyte response. Store aliquots at −20°C.
• Sample spiking: Add a fixed volume of IS working solution to biological samples prior to extraction (e.g., 10–50 µL per 0.1–0.5 mL matrix), maintaining consistent final organic content.
• Extraction: Perform protein precipitation or SPE suitable for glucuronides. Avoid extreme pH.
• LC–MS/MS: Monitor MRM transitions corresponding to the deuterated analog and the native analyte; optimize collision energies to preserve the glucuronide moiety or to track diagnostic fragments as required.

Note: All parameters above are general guidance and not item-specific specifications. For regulated studies, establish and document method-specific conditions.

Biological Roles

General background (literature):

• Nicotine N-β-D-glucuronide is a phase II metabolic conjugate formed by UDP-glucuronosyltransferases (UGTs), which increases hydrophilicity and facilitates renal excretion.
• In humans, nicotine undergoes multiple metabolic transformations (e.g., oxidation to cotinine; N-oxidation; glucuronidation). N-glucuronidation decreases membrane permeability and receptor binding potential relative to nicotine.
• The glucuronide conjugate serves as a biomarker of nicotine metabolism and exposure, complementing cotinine and its glucuronides in biomonitoring.
• Deuterium-labeled analogs are not endogenous; they function as internal standards for accurate quantitation in mass spectrometric assays through isotopic dilution.

Biochemical considerations:

• UGT isoforms implicated in nicotine N-glucuronidation include several hepatic and extrahepatic enzymes (literature reports vary by species and tissue). The conjugate is typically found in urine and, to a lesser extent, in plasma.
• The β-D-glucuronide linkage can be cleaved enzymatically by β-glucuronidase, a feature exploited in analytical hydrolysis protocols to back-calculate total nicotine-related metabolites.

Note: The above represents general biological knowledge for nicotine glucuronidation and does not ascribe any therapeutic or clinical properties to this product.

Buffer Applications

As a small-molecule analytical standard rather than a buffering agent, this compound is not used to establish buffer capacity.

Practical notes for solution preparation (general):

• Prepare standards and QCs in water or aqueous buffer compatible with LC–MS (e.g., 5–10 mM ammonium acetate or formate, pH ~3–6). Avoid high-ionic-strength buffers that suppress ionization.
• For enzymatic studies (e.g., β-glucuronidase hydrolysis), typical biological buffers (phosphate, acetate) at pH 5–7 are used; verify that buffer components do not interfere with MS detection.
• Filter through 0.2 µm low-binding membranes when particulate contamination is a concern; use polypropylene or low-adsorption vials to limit analyte loss.

Green Alternatives

For this isotopically labeled analytical standard, “green alternatives” primarily concern solvent and workflow choices rather than replacement of the analyte itself.

Greener method considerations (general):

• Mobile phases: Favor aqueous buffers with volatile, low-toxicity additives (e.g., ammonium formate/acetate) over non-volatile salts; use lower organic content where chromatographic performance allows.
• Organic modifiers: Between acetonitrile and methanol, consider methanol for renewability; however, acetonitrile can reduce backpressure and energy consumption via shorter runs. Optimize to minimize total solvent use.
• Sample prep: Employ protein precipitation with ethanol or isopropanol as alternatives to acetonitrile where compatible. Miniaturize extraction volumes and adopt 96-well formats to reduce waste.
• Chromatography: Utilize sub-2 µm or core–shell columns to shorten runtimes and cut solvent consumption without sacrificing resolution, particularly for polar glucuronides which often require only modest organic content.

Tradeoffs (general):

• Methanol is renewable but more viscous than acetonitrile, potentially increasing pump energy and run time.
• Aqueous-rich methods reduce flammability and VOC exposure but may demand HILIC or ion-pair strategies; avoid non-volatile ion-pairs to keep MS compatibility.

Note: There is no practical "green alternative" to the labeled standard itself for quantitative LC–MS; rather, implement greener solvents and scaled-down protocols.

Pharmaceutical Uses

This product is supplied strictly for research use only and is not intended for human or veterinary use, diagnostic procedures, or as an active pharmaceutical ingredient.

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

• Bioanalytical internal standard for LC–MS/MS quantification of nicotine glucuronide in nonclinical and clinical sample testing labs (method development/validation). Supports pharmacokinetic profiling and exposure assessment workflows (no therapeutic claims).
• Reference material for metabolite identification (MetID) studies and for evaluating metabolic pathways involving UGT-mediated conjugation.
• System suitability and calibration verification material for regulated and non-regulated assays, provided that batch-specific purity and isotopic enrichment meet method criteria as documented on the CoA.

Regulatory notes (general):

• Not a compendial standard; no pharmacopeial monograph is typically available for this isotopologue. Laboratories should establish in-house reference standard qualification procedures consistent with internal SOPs and applicable guidance (e.g., FDA/EMA bioanalytical method validation) without implying any clinical use of this product.

Physical Properties

Item-specific physicochemical values are not provided.

• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
• Appearance: 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.
• Solubility (general/literature): Nicotine N-glucuronide conjugates are highly polar and typically freely soluble in water and aqueous buffers; sparingly soluble in most nonpolar organic solvents. They are commonly reconstituted in water, aqueous methanol, or aqueous acetonitrile for LC–MS.
• Ionization/acid–base behavior (general/literature): N-glucuronides often exist as zwitterions or salts; the glucuronic acid moiety (carboxylate) is deprotonated near neutral pH, enhancing aqueous solubility. The nicotine nitrogen is neutral in the N-glucuronide but can form salts depending on counterions.
• Partitioning (general/literature): Very low logP and high logD(aq) favoring the aqueous phase; limited membrane permeability.

Handling note: Being a deuterated standard, minimize evaporative losses and prolonged exposure to ambient moisture to preserve concentration and isotopic integrity.

Quality and Grades

• Grade/purity (item-specific): Not specified for this item; refer to CoA/Spec Sheet.

Context for this product class (general):

• Isotopically labeled metabolite standards are typically supplied at high chemical purity and high isotopic enrichment to ensure accurate quantitation by LC–MS/MS. The exact percent chemical purity and percent deuterium incorporation can significantly affect response factors and should be verified on the CoA.
• Counter-ions/form: These standards may be supplied as free acids, ammonium salts, or other salt forms to optimize solubility and stability. The salt/form and water content should be confirmed on the CoA.
• Implications for analytical work: Lower UV background and minimal inorganic residue are preferred for LC–MS; verify residual solvent profile and inorganic content on the CoA if relevant to regulated workflows.
• Traceability: For quantitative method validation, document batch-specific purity, labeling pattern, and concentration of stock solutions prepared gravimetrically or volumetrically. Retain the CoA and any isotopic purity certificate for audit trails.

Reaction and Applications

This product is primarily used as an analytical internal standard rather than as a synthetic reagent.

Analytical applications (general/literature):

• LC–MS/MS internal standard for quantifying nicotine N-glucuronide in biological matrices (urine, plasma) during exposure assessment, pharmacokinetic, or metabolism studies (research use only).
• Method validation: Suitable for calibration curves, isotopic dilution, spike-recovery, and matrix effect evaluation. Deuterium labeling facilitates co-elution with minimal chromatographic isotope effect.
• Stability assessment: Useful for evaluating enzymatic hydrolysis susceptibility (β-glucuronidase challenge) and for recovery studies across sample prep workflows (protein precipitation, SPE).

Enzymology/metabolism studies (general):

• Reference substrate/product marker in UGT-mediated nicotine metabolism investigations.
• Control analyte in assays assessing deconjugation by β-glucuronidase and the kinetics of back-conversion to nicotine under stressed conditions.

Notes for use:

• Employ low-adsorption plastics or silanized glass to mitigate surface binding of polar glucuronides.
• Prepare fresh working solutions or store aliquots at low temperature to minimize degradation or isotopic exchange; verify by system suitability injections.

Reaction Conditions

This product is used primarily as an analytical standard rather than as a reagent in synthetic reactions. Accordingly, there are no canonical “reaction conditions” for its use.

General guidance for analytical use (literature-based):

• LC–MS/MS: Electrospray ionization in negative or positive mode can detect glucuronides; many methods favor negative ESI due to the carboxylate group, but N-glucuronides of amine-containing analytes are also detected in positive mode depending on adduct formation. Optimize source parameters for minimal in-source fragmentation of the glucuronide.
• Chromatography: Reverse-phase with aqueous-rich starting conditions or HILIC for enhanced retention of highly polar analytes. Typical column temps 25–40°C; flow 0.2–0.5 mL/min on 2.1 mm ID columns (general values).
• Sample preparation: Protein precipitation using cold methanol or acetonitrile (3–4× volume) followed by dilution; or SPE using mixed-mode cation/anion-exchange cartridges tailored for glucuronides.

These conditions are general literature guidance for glucuronide analytics and are not item-specific specifications. Confirm suitability with small-scale tests and system suitability checks.

Safety and Handling

• GHS classification (item-specific): 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 (not a substitute for the SDS):

• Potential hazards: Nicotine derivatives can exhibit toxicity. Although glucuronidation typically reduces biological activity relative to parent nicotine, treat this material as toxic and harmful if swallowed, inhaled, or in contact with skin. Avoid aerosol formation.
• PPE: Use lab coat, safety glasses, and appropriate chemical-resistant gloves (e.g., nitrile). Work in a fume hood or well-ventilated area, especially when preparing stock solutions for LC–MS.
• Incompatibilities: Avoid strong oxidizers and strong acids/bases that can hydrolyze the glucuronide linkage. Maintain solutions near neutral pH unless hydrolysis is intended for analytical purposes.
• First aid (general): In case of skin contact, wash with soap and water. If eye contact occurs, rinse cautiously with water for several minutes and seek medical attention. If inhaled, move to fresh air. If ingested, rinse mouth; obtain medical attention.
• Spill/cleanup: Absorb small spills with inert material, avoid dust/solution aerosols, and dispose of according to institutional and local regulations.
• SDS: Always consult the product-specific SDS for authoritative hazard and response information.

Solvent Selection

This compound is a highly polar N-glucuronide, typically favoring aqueous media.

• Polarity/miscibility (general/literature):

  • Highly water soluble; miscible with aqueous buffers.
  • Compatible with typical LC–MS mobile phases: water with 0.1% formic acid or 5–10 mM ammonium formate/acetate; organic modifiers such as methanol or acetonitrile are used in gradients.
  • Poor solubility expected in nonpolar solvents (hexane, toluene) and limited in moderately polar aprotic solvents unless water is present.

• Practical guidance:

  • Prepare stock solutions in water or aqueous methanol/acetonitrile (e.g., 50:50, v/v) to improve stability and handling; filter if necessary through low-binding membranes.
  • For SPE method development, hydrophilic interaction chromatography (HILIC) or mixed-mode ion-exchange phases often retain glucuronides effectively.
  • Avoid high-pH media that can promote glucuronide hydrolysis; keep within pH 3–8 unless hydrolysis is intended.

• Comparison (general):

  • Water vs. MeOH: Water maximizes solubility and native ionization; methanol-water mixes can aid wetting and injection reproducibility.
  • ACN vs. MeOH as modifier: ACN offers lower viscosity and sharper LC peaks, while MeOH may enhance ESI response for some glucuronides. Optimize empirically for your instrument.

Storage and Reconstitution

• Storage conditions (item-specific): Store at −20°C.
• Shipping (item-specific): Shipped in an ice chest with ice pads to maintain a low temperature during transit.

Reconstitution and handling (general guidance):

• Solvents: Use high-purity water or LC–MS-grade aqueous mixtures (e.g., water:methanol or water:acetonitrile) for preparing stock and working solutions. Filter if particulates are visible.
• Concentration: Not specified for this item; refer to CoA/Spec Sheet. Prepare concentrations appropriate for your assay and instrument response.
• Aliquoting: Upon first use, prepare single-use aliquots to minimize freeze–thaw cycles and potential hydrolysis of the glucuronide linkage.
• Stability: Keep solutions cold and protected from light where possible. Avoid prolonged exposure to high pH or elevated temperatures that can promote deconjugation.
• Containers: Use low-bind polypropylene or glass vials with PTFE-lined caps. Verify adsorption losses by recovery checks when working at low ng/mL levels.

Always consult the product’s CoA and SDS for the most accurate, lot-specific information on composition, concentration (if supplied as solution), and stability.

Structure and Identity

This product is a deuterium-labeled phase II conjugate of nicotine, where the tertiary amine of nicotine is N-glucuronidated to yield an N-β-D-glucuronide. The “-d” indicates deuterium incorporation for use as an isotopically labeled internal standard.

• SKU: N1444242
• Product name: Nicotine N-β-D-glucuronide-d
• CAS: 329002-74-6
• Category: Life science research reagent; metabolite analytical standard (research use only)
• Molecular formula (item-specific): Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight (item-specific): Not specified for this item; refer to CoA/Spec Sheet.
• SMILES (item-specific): Not specified for this item; refer to CoA/Spec Sheet.
• InChIKey (item-specific): Not specified for this item; refer to CoA/Spec Sheet.

Structural features (general/literature):

• Core scaffold: Nicotine (a pyridine ring fused to a N-methyl-2-pyrrolidine motif) bearing a quaternizable tertiary amine.
• Conjugation: N-glucuronidation at the nicotine nitrogen, linked via a β-glycosidic bond to D-glucuronic acid (typical for phase II metabolism of tertiary amines).
• Isotopic label: One or more hydrogen atoms replaced by deuterium (positioning varies by supplier; consult CoA for exact labeling pattern).
• 2D description: A six-membered aromatic pyridine ring attached to a saturated five-membered ring containing a tertiary amine; the nitrogen carries a glucuronide substituent comprising a pyranose (D-glucuronic acid) with carboxylate functionality.

Note: Exact isotopic enrichment and labeling positions are item-specific and must be confirmed from the CoA/Spec Sheet.

Synthetic Utility

This compound is not typically used as a building block in synthetic chemistry. Its primary value lies in analytical workflows. However, understanding its functional groups informs stability and handling.

Functional group features (general):

• β-D-Glucuronide (pyranose carboxylate) linked to a tertiary amine nitrogen on the nicotine scaffold. The glycosidic linkage and carboxylate confer high polarity and susceptibility to hydrolysis under strongly acidic or basic conditions.
• Isotopic deuterium labeling offers a mass shift useful for internal standardization but is not intended for isotopic exchange reactions.

Implications for chemists:

• Avoid conditions that cleave N–glucuronide bonds (strong base, strong mineral acids, elevated temperatures) when preparing and storing analytical solutions.
• When exploring metabolism-mimicking syntheses, chemical or enzymatic N-glucuronidation methods and protecting-group strategies for glucuronic acid are relevant; however, the labeled product should not be used as a precursor for further transformations due to cost and potential isotope scrambling.

Target Specificity

Not applicable. This is a small-molecule analytical standard, not an antibody or affinity reagent. No antigen, epitope, species reactivity, clone, or isotype information is relevant to this product.