Ethanol Assay Kit

Catalog No. ABIN1000261

Product Details

Target: Ethanol

Reactivity: Chemical

Application: Biochemical Assay (BCA)

Sample Type: Beverages, Serum, Plasma

Specificity: 0.04 %

Characteristics: Sensitive and accurate. Detection range 0.04 - 4 % alcohol in 96-well plate assay.
Convenient and high-throughput. The procedure involves adding a single working reagent, incubation for 8 min, adding a Stop Reagent, and reading the optical density. Can be readily automated as a high- throughput 96-well plate assay for thousands of samples per day.
Versatility. Assays can be executed in 96-well plate or cuvet.

Components: Reagent A: 50 mL. Reagent B: 50 mL. 10% TCA: 50 mL. Standard: 2 mL 10% (v/v) ethanol.

Material not included: Pipeting (multi-channel) devices. Clear-bottom 96-well plates (e.g. Corning Costar) and plate reader. Centrifuge tubes, table centrifuge, cuvets and spectrophotometer.

Application Details

Application Notes: Ethanol determination in alcohol containing samples such as beverages (e.g. wine, beer) and yeast cultures. For samples containing less than 0.1% alcohol such as serum or plasma, Ethanol Assay Kit is recommended.

Comment:

If sample contains glucose or glycerol, use BioAssay Systems' specific ethanol assay kit (ECET-100). For samples containing only sugars (e.g. glucose), use Saccharide Removal

Protocol: Procedure using 96-well plate:
1. Prepare 600 µL 2% Premix by mixing 120 µL 10% Standard and 480 µL distilled water. Transfer 100 µL standards and samples into wells of a clear bottom 96-well plate
2. Add 100 µL Reagent A quickly using a multi-channel pipettor. Tap plate lightly to mix.
3. Incubate 8 to 30 min at room temperature. The reagent color changes from yellow to visibly bluish in wells 1-4. Add 100 µL Stop Reagent B quickly using a multi-channel pipettor. Tap plate to mix.
4. Read OD at 570-600nm (peak 580nm).

Procedure using cuvette:
1. Prepare 2%, 1%, 0.5% standards and use distilled water as blank control. Transfer 400 µL diluted Standards and 400 µL samples to1.5-mL centrifuge tubes.
2. Add 400 µL Reagent A quickly to each tube and vortex briefly to mix.
3. Incubate 8 to 30 min at room temperature. Add 400 µL Reagent B quickly and mix briefly.
4. Transfer to cuvettes and read OD at 570-600nm (peak 580nm). Note: for the cuvette assay, it is recommended that an interval be applied between additions, e.g., add Reagent A to Tube 1 and 1 min later to Tube 2 etc. After the incubation step is completed, add the Stop Reagent B to Tube 1 and 1 min later to Tube 2 etc. This will ensure identical incubation time between tubes.

Calculation of Results:

Subtract blank OD (water, #8) from the standard OD values and plot the OD against standard alcohol concentrations. Determine sample ethanol concentration from the standard curve.
Conversions: 1% (v/v) ethanol equals 170 mM or 785 mg/dL.

Restrictions: For Research Use only

Handling

Storage: 4 °C

References

Pisu, Mostallino, Dore, Maciocco, Secci, Serra: "Effects of voluntary ethanol consumption on emotional state and stress responsiveness in socially isolated rats." in: European neuropsychopharmacology : the journal of the European College of Neuropsychopharmacology, Vol. 21, Issue 5, pp. 414-25, (2011) (PubMed).

Kim, Im, Ko, Chin, Kim, Lee, Bai: "Raw starch fermentation to ethanol by an industrial distiller's yeast strain of Saccharomyces cerevisiae expressing glucoamylase and α-amylase genes." in: Biotechnology letters, Vol. 33, Issue 8, pp. 1643-8, (2011) (PubMed).

Wang, Chung, Rhoads: "Altered pattern of Na,K-ATPase activity and mRNA during chronic alcohol consumption by juvenile and adolescent rats." in: Cellular and molecular neurobiology, Vol. 29, Issue 1, pp. 69-80, (2009) (PubMed).

Alam, Kabbashi, Hussin: "Production of bioethanol by direct bioconversion of oil-palm industrial effluent in a stirred-tank bioreactor." in: Journal of industrial microbiology & biotechnology, Vol. 36, Issue 6, pp. 801-8, (2009) (PubMed).

Lin, Lu, Zhang, Walter, Dang, Wan, Tao, Qian, Zhao, Boeke, Berger, Zhu: "Protein acetylation microarray reveals that NuA4 controls key metabolic target regulating gluconeogenesis." in: Cell, Vol. 136, Issue 6, pp. 1073-84, (2009) (PubMed).

Biggio, Gorini, Caria, Murru, Sanna, Follesa: "Flumazenil selectively prevents the increase in alpha(4)-subunit gene expression and an associated change in GABA(A) receptor function induced by ethanol withdrawal." in: Journal of neurochemistry, Vol. 102, Issue 3, pp. 657-66, (2007) (PubMed).

Khanna, Kan, Fang, Xie, Underwood, Jain, Williams, Finkel: "Inducible nitric oxide synthase attenuates adrenergic signaling in alcohol fed rats." in: Journal of cardiovascular pharmacology, Vol. 50, Issue 6, pp. 692-6, (2007) (PubMed).

Target Details

Target: Ethanol

Background: Quantitative determination of ethanol by chemical colorimetric (580nm) method.
Procedure: 10 min.

Alcoholic drinks are among the daily consumed beverages. Studies have shown heavy alcohol consumption may lead to various forms of liver diseases and to increased mortality rates. Quantitative determination of alcohol (ethanol, C2H5OH) finds applications in basic research, drug discovery, clinic studies and winery. Simple, direct and automation-ready procedures for measuring ethanol concentration are very desirable. This ethanol assay kit is based on an improved dichromate method, in which dichromate is reduced by ethanol to a bluish chromic (Cr 3+ ) product. The intensity of color, measured at 580 nm, is a direct measure of the alcohol concentration in the sample. The optimized formulation substantially reduces interference by substances in the raw samples and exhibits high sensitivity.