Dehydroascorbic Acid (DHA) Content Assay Kit (UV Micro Method)

Cat. No.: D1521839

CALIDAD & PUREZA BioReagent

Descripción general

Ascorbic acid (AsA) serves as a key physiological indicator in plant cells. Changes in its content, redox status (AsA/DHA ratio), and the activities of enzymes involved in its synthesis and metabolism are associated with plant responses to a range of environmental stresses. Dehydroascorbic acid (DHA) is the reversibly oxidized form of AsA and, together with ascorbic acid, constitutes a redox system in organisms, functioning as an electron acceptor. The detection principle of this kit is based on the specific and complete reduction of DHA in the sample to AsA by the reductant dithiothreitol (DTT). The DHA content is then calculated by measuring the rate of AsA generation in the system.

Especificaciones

Especificaciones y pureza

BioReagent

Grado

BioReagent

Aplicación

Cell Metabolism

Tipo de muestra

Tejidos vegetales

Rango de detección

0.01-5 μmol/mL

Método de detección

Colorimetric Method

Detection instrument

Microplate reader

Detection wavelength

265nm

Almacenamiento y envío

Condiciones de almacenamiento de almacenamiento

Protected from light,Store at -20°C

Enviado en

Ice chest + Ice pads

Estabilidad y almacenamiento

Store at -20℃ for 6 months. Store in the dark.

Application Protocols

Item-specific tested applications, plate formats, step-by-step procedures, and calculation worksheets are not specified for this item; refer to the kit’s IFU/CoA for authoritative protocols.

• General workflow outline (literature/general)
  • Prepare standards and samples in assay buffer; protect from light.
  • If required, measure baseline absorbance (ascorbate present).
  • Add selective reagent to convert DHA to ascorbate or to mask native ascorbate (method-dependent); incubate for the prescribed time.
  • Measure UV absorbance in a microplate reader at the specified wavelength; subtract blanks and calculate concentrations from the standard curve.
  • Validate with matrix spikes/recoveries and replicate wells.

Ensure instrument settings (pathlength correction, temperature control, shaking) and plate type match the IFU. Dispose of waste per institutional and local regulations.

Biological Roles

This section describes the biology of the analyte (DHA) for context; it does not describe any medical use of the kit.

• Vitamin C redox couple (literature/general)
  • Ascorbate (AscH−) is a major aqueous antioxidant. Dehydroascorbic acid (DHA) is its two-electron oxidized form. The AscH−/DHA couple buffers reactive oxygen species by donating electrons to radicals and oxidants.
• Transport and metabolism (literature)
  • Ascorbate typically enters cells via SVCT transporters, whereas DHA can enter via GLUT transporters due to structural resemblance to glucose; intracellular DHA is rapidly reduced back to ascorbate by glutathione-dependent and enzymatic systems (e.g., thioredoxin/glutaredoxin pathways).
  • DHA is chemically labile, hydrating in aqueous solution and degrading to 2,3-diketo-L-gulonic acid if not promptly reduced.
• Experimental relevance (literature/general)
  • The DHA:ascorbate ratio serves as a proxy for cellular redox state in models of oxidative stress, hypoxia, metabolism, and food processing stability.
  • Accurate DHA quantification requires controlling ex vivo oxidation; rapid quenching, low temperature, and light protection are critical.

Note: Biological insights are provided for research planning only. Item-specific performance for biological matrices is not specified for this item; refer to CoA/Spec Sheet.

Buffer Applications

This product is an analytical kit, not a buffering agent. Traditional buffer system design (pH ranges, recipes, electrophoresis buffers) is not directly applicable.

• Practical note (general)
  • The kit will include or specify assay buffers appropriate for DHA stabilization and UV measurement. Exact compositions, pH targets, and ionic strengths are not specified for this item; refer to CoA/Spec Sheet.
  • When preparing user-supplied buffers for sample extraction, avoid components with strong UV absorbance near ~260–270 nm (e.g., nucleotides, certain aromatics) and match buffer composition across standards and samples to minimize baseline bias (literature guidance).

Green Alternatives

This UV microplate kit aligns with greener analytical principles by avoiding large volumes of hazardous organic solvents and by enabling miniaturized measurements.

• Greenness considerations (general)

  • Microplate UV format minimizes reagent consumption and waste compared with cuvette or HPLC methods.
  • Aqueous buffers are typically used, reducing reliance on chlorinated solvents or acetonitrile (common in derivatization/HPLC assays).
  • Room-temperature operation often suffices, lowering energy demand.

• Comparison to common alternatives (literature/general)

  Greener attribute | UV microplate DHA kit | Derivatization + HPLC ------------------|-----------------------|----------------------- Solvent use | Predominantly aqueous | Often organics (ACN/MeOH) Throughput | High (96/384 wells) | Moderate Waste volume | Low per sample | Higher per run Specificity | Moderate–high (method dependent) | High with separation Sensitivity | Adequate for many matrices | High (fluorescence/MS)

• Trade-offs

  • UV absorbance can suffer from matrix interference; careful sample cleanup and blanks are needed.
  • HPLC methods may better resolve complex matrices but at higher environmental and cost footprints.

Item-specific reagent compositions and disposal recommendations are not specified for this item; refer to CoA/Spec Sheet and local regulations.

Pharmaceutical Uses

No direct pharmaceutical formulation or excipient role applies to this assay kit.

• Relevant research/industry contexts (general)
  • Can be used in research laboratories and quality-control investigations to assess DHA/ascorbate content in biological samples, raw materials, nutraceuticals, and processed foods (non-clinical, research use only).
  • Method development aid for stability studies of vitamin C in formulations or during manufacturing processes, where a rapid, small-volume UV assay is advantageous.

Item-specific regulatory listings (e.g., pharmacopeial status, validation packages, reportable ranges) are not specified for this item; refer to CoA/Spec Sheet. No medical or clinical diagnostic use is intended or implied.

Physical Properties

Because this is a multi-component assay kit, single-chemical physical constants do not apply to the entire product.

• Item-specific physical specs (kit-level): Not specified for this item; refer to CoA/Spec Sheet.
• Typical properties of the analyte and assay readout (literature/general)
  • Dehydroascorbic acid (DHA) MW (literature): ~174.11 g/mol (C6H6O6). For reference, reduced ascorbic acid is ~176.12 g/mol (C6H8O6).
  • UV behavior (literature): Ascorbate exhibits a UV absorption band near ~265 nm in aqueous solution; DHA has substantially weaker absorbance at this wavelength. Many DHA assays reduce DHA to ascorbate and quantify by ΔA265–270 (literature principle).
  • Aqueous behavior (literature): DHA is unstable in water, tending to hydrate and further degrade to 2,3-diketo-L-gulonic acid; stability improves at low temperature and low pH.
  • Solubility (literature): DHA and ascorbate are highly water-soluble; solubility decreases markedly in nonpolar solvents.

Practical note: Exact buffer compositions, molarities, extinction coefficients, and microplate pathlength corrections used in this kit are not specified for this item; refer to CoA/Spec Sheet and the included protocol for definitive parameters.

Quality and Grades

• Item-specific quality designation
  • Grade/Purity: BioReagent (as provided). This denotes suitability for common biochemical workflows, where background absorbance, trace contaminants, and stability meet research-grade expectations for analytical assays.
• Implications of BioReagent grade (general)
  • Low background in UV-visible range to minimize baseline drift in microplate measurements.
  • Components prepared to minimize interfering impurities that could reduce/oxidize DHA/ascorbate nonspecifically.
  • Packaging that supports light and moisture protection, essential for ascorbate/DHA chemistry.
• What is and is not specified for this item
  • Item-specific assay performance claims (e.g., LOD/LOQ, linear range, accuracy, precision), stabilizers, and component lot specifications: Not specified for this item; refer to CoA/Spec Sheet.
• Practical quality considerations
  • Verify instrument baseline with provided blanks to assess any intrinsic absorbance of kit buffers (good UV transparency is expected for BioReagent grade).
  • Use the supplied standard(s) to construct a standard curve each run; BioReagent grade supports reproducibility across plates and days when handled per IFU.
  • Light protection and cold-chain integrity during shipping/storage are integral parts of quality for redox-sensitive assays.

Recommendation: Document lot numbers and retain CoA/Spec Sheet to correlate assay performance with component specifications.

Reaction and Applications

• Intended application (from product name/category)
  • Quantitative determination of dehydroascorbic acid (DHA) content using a UV microplate method. Suitable for life-science and analytical workflows; for research use only.
• Assay principle (literature/general)
  • Many DHA assays operate by selectively reducing DHA to ascorbate and measuring the increase in absorbance near ~265–270 nm attributable to ascorbate; the difference before and after reduction provides DHA content. Alternatively, selective masking of pre-existing ascorbate may be used, followed by total ascorbate determination (literature approaches).
• Typical applications (general)
  • Assessing redox state of vitamin C pools (ascorbate vs DHA) in cell lysates, tissues, culture media, fermentation broths, plant extracts, and food/beverage samples.
  • Monitoring oxidative stress interventions in biochemical experiments by tracking DHA fraction.
  • Quality assessment of sample handling and storage (ascorbate oxidation) during method development.
• Practical tips (general)
  • Work quickly at low temperature and protect from light to limit ex vivo oxidation of ascorbate to DHA.
  • Clarify turbid/particulate samples via centrifugation/filtration to avoid scattering artifacts in UV micro readings.
  • Use matched buffers and matrix spikes to check recovery; construct a fresh standard curve every run.

Item-specific workflow steps, reagent identities, and calculation templates are not specified for this item; consult the kit IFU/CoA.

Reaction Conditions

This section does not describe a synthetic reaction but provides general assay-condition guidance for UV-based DHA determinations. Item-specific protocols, reagent identities, and parameters are not specified for this item; refer to CoA/Spec Sheet/IFU for definitive instructions.

• General assay conditions (literature/general)
  • Temperature: Ambient operation is common; colder temperatures are used during sample prep to minimize ascorbate oxidation.
  • Light: Protect from light throughout handling to preserve redox state.
  • pH: Mildly acidic conditions are often employed to stabilize ascorbate/DHA and to limit side reactions.
  • Detection: Microplate UV absorbance near ~265–270 nm is commonly used for ascorbate following selective reduction of DHA (principle-level guidance).
  • Timing: Rapid processing minimizes ex vivo conversion; incubations for reduction/masking typically range from minutes to tens of minutes depending on reagent chemistry (literature).
• Quality controls (general)
  • Run blanks, standards, and matrix spikes; assess linearity and recovery.
  • Include duplicate wells and, if possible, before/after reduction measurements to calculate DHA by difference.

Again, use the official kit instructions for exact volumes, incubation times, and wavelengths.

Safety and Handling

• Regulatory and GHS
  • Signal Word, H-Statements, GHS Classification, Pictograms: Not specified for this item; refer to SDS for each kit component.
  • Research Use: For research use only.
• General hazards for DHA assay kits (general guidance)
  • Kits may include acids/buffers, reducing agents (e.g., trialkylphosphines or thiols), and preservatives. Such reagents can be irritants, corrosive, or harmful if inhaled/ingested.
  • UV micro methods require microplate readers; avoid direct eye/skin exposure to UV sources and handle microplates carefully to prevent spills.
• PPE and engineering controls
  • Wear lab coat, safety glasses, and disposable nitrile gloves. Work in a well-ventilated area; use a fume hood when handling volatile or odorous reagents.
• Storage incompatibilities (general)
  • Keep reducing agents away from strong oxidizers; segregate acids from bases and cyanides; avoid metal contamination if the kit contains phosphine reductants.
• First-aid overview (general)
  • Skin/eye contact: Rinse with water for at least 15 minutes; remove contaminated clothing.
  • Inhalation: Move to fresh air; seek medical advice if symptoms persist.
  • Ingestion: Rinse mouth; do not induce vomiting; seek medical attention.
• Waste
  • Collect aqueous assay waste containing reductants/preservatives in appropriate containers; dispose per institutional and local regulations.

Always defer to the SDS and the kit IFU for authoritative safety guidance.

Solvent Selection

This is an assay kit rather than a synthetic reagent; solvent selection primarily concerns sample preparation and matrix compatibility.

• General solvent/matrix guidance (literature/general)
  • Aqueous buffers at controlled pH are typically used to preserve DHA and to enable the UV readout after reduction to ascorbate.
  • Organic co-solvents should be minimized because they can affect extinction coefficients and baseline absorbance; if required, keep below a few percent v/v and match in standards and samples.
  • For solid or lipid-rich samples (e.g., foods), aqueous extraction with protein precipitation (e.g., with metaphosphoric acid or TCA, literature) is common to stabilize ascorbate/DHA and remove turbidity before microplate reading.
• When to choose alternatives
  • If the sample is highly colored or turbid and interferes at ~260–270 nm (literature), consider parallel clarification (centrifugation/filtration) or alternative detection chemistries (e.g., fluorescence- or derivatization-based methods) outside the scope of this specific kit.

Item-specific extraction buffers, allowable solvent percentages, and compatible matrices are not specified for this item; refer to CoA/Spec Sheet and the kit instructions for definitive guidance.

Storage and Reconstitution

• Item-specific storage/shipping
  • Storage Conditions: Protected from light, Store at -20°C (as provided).
  • Shipped In: Ice chest + Ice pads (as provided) to maintain cold chain.
• Stability considerations (general)
  • Many DHA/ascorbate-related reagents are light- and temperature-sensitive. Minimize freeze–thaw by aliquoting reconstituted solutions when permitted.
  • Keep containers tightly closed and purge headspace with inert gas if recommended by the IFU (item-specific guidance not provided here).
• Reconstitution and preparation
  • Exact reconstitution solvents, volumes, and working-solution stability are not specified for this item; refer to CoA/Spec Sheet.
  • General practice: Use high-purity water or the supplied buffer. Bring reagents to the recommended temperature before mixing; dissolve completely with gentle inversion, avoiding bubble formation that can affect UV readings.
• In-use handling
  • Protect plates and reagents from ambient light; use amber tubes/wraps.
  • Prepare fresh standards each session; discard expired working solutions.

Always follow the official kit IFU for reconstitution instructions, allowable storage durations after opening, and component-specific stability.

Structure and Identity

This product is an analytical kit designed to quantify dehydroascorbic acid (DHA) via a UV microplate method. It is not a single chemical substance and therefore has no unique CAS/SMILES/InChIKey for the entire kit.

• Item-specific identifiers
  • SKU: D1521839
  • Product Name: Dehydroascorbic Acid (DHA) Content Assay Kit (UV Micro Method)
  • CAS, CID, InChIKey, SMILES, Molecular Formula, Molecular Weight: Not specified for this item; refer to CoA/Spec Sheet.
• What the kit measures (general/literature context)
  • Analyte: dehydroascorbic acid (oxidized form of vitamin C).
  • Typical structural features of DHA (literature): bicyclic lactone/lactol framework derived from L-ascorbic acid; enediol system oxidized to a diketone in DHA; readily hydrates in water and can tautomerize.
  • 2D structure description (literature): a five-membered lactone ring fused to an enediol-derived system; in DHA, the C2–C3 enediol of ascorbate is oxidized to carbonyls, diminishing the UV chromophore until reduced back to ascorbate.
• Kit composition note
  • Exact component list (e.g., buffers, reductant, standards, stop reagents) is not specified for this item; refer to CoA/Spec Sheet.

Summary: Use this kit to selectively quantify DHA based on a UV readout after an analyte-specific reaction. Structural identifiers do not apply to the full kit.

Synthetic Utility

Not applicable. This product is an analytical assay kit and is not used as a building block or reagent in chemical synthesis.

• For users interested in chemistry of the analyte (literature/general)
  • DHA participates in reversible redox chemistry with ascorbate and can undergo nucleophilic additions (e.g., amines to carbonyl centers) and hydration; however, these reactions are typically undesirable during analysis and are controlled by buffer composition, pH, and temperature.

Focus instead on the sections detailing assay applications, safety, and storage.

Target Specificity

• Intended analyte: Dehydroascorbic acid (DHA), oxidized form of vitamin C. This contextual statement is based on the product name.
• Item-specific details such as cross-reactivity, interference profile, linear range, LOD/LOQ, and matrix compatibility are not specified for this item; refer to CoA/Spec Sheet and the kit instructions for validated performance characteristics.