QuantiChrom™ Total Carbohydrate Assay Kit

Protocol SDS

Application

• For quantitative determination of total carbohydrates in food, beverage, and biological samples (e.g. serum, plasma, etc).

Key Features

• Non-radioactive assay.
• Sensitive and accurate. Linear detection range in 96-well plate: 0.02 to 5 mM glucose equivalents for colorimetric assays.
• Fast and convenient. The procedure involves 15 minutes of incubation time and minimal sample preparation.

Method

• OD490nm

Samples

• Food, beverage, biological samples (e.g. serum, plasma, etc).

Species

• All

Procedures

• 15 min

Size

• 100 tests in a 96-well plate

Detection Limit

• 0.02 mM

Shelf Life

• 6 months

More Details

CARBOHYDRATES are made up of carbon, hydrogen, and oxygen. A carbohydrate may exist in its simplest form, a monosaccharide, or as larger polymers called polysaccharides (e.g. starch, glycogen, and cellulose). Carbohydrates are essential constituents of living things, and in the form of glucose, serve as the main source of energy for metabolism. They are also commonly found in many fruits and vegetables as simple or complex sugars. BioAssay Systems’ QuantiChrom™ Total Carbohydrate Assay Kit first hydrolyzes polysaccharides to monosaccharides, which are then converted to furfural compounds that react with the Detection Reagent. The color intensity of the chromagen, measured at 490 nm, is directly proportional to the total carbohydrate concentration (glucose equivalents) in the sample.

Does the Detection Reagent react with all carbohydrates? If so, is it to the same degree?

The assay reacts with all carbohydrates (monosaccharides, disaccharides, etc.). The absorbance of equimolar sugars will differ and therefore it is important that the concentration of carbohydrates in a sample is arbitrarily expressed as the equivalents of one sugar (D-glucose in the case of TCRB-100).

Can the entire assay be performed in a microplate?

The assay was designed to be performed in microcentrifuge tubes and then transferred to a microplate for reading. However, as long as the material of the microplate is not sensitive to sulfuric acid (e.g. polystyrene), the assay should be able to be done from start to finish in a microplate. It is recommended to test the resilience of plastics to sulfuric acid prior to performing the assay by adding 100 μL of concentrated sulfuric acid (95%) and heating at 90°C for 5 minutes.

During the heating step, cover the plate with a sealing film to prevent loss of solution. Upon the addition of the Detection Reagent to each well, it is necessary to pipette up and down multiple times to mix well. After the 10 minute incubation with the Detection Reagent, transfer 100 μL from each well to separate wells in a clear, flat bottom 96-well plate.

If my sample has a mixture of carbohydrates, how accurate are the results?

Since the absorbance of the colored product varies in response to different sugars, this assay may over- or underestimate carbohydrate content in mixtures or biological samples.

For more detailed product information and questions, please feel free to email or call us at 1-510-782-9988 x 2. For more general information regarding our assays, please refer to Technical Support.

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If you or your labs do not have the equipment or scientists necessary to run this assay, BioAssay Systems can perform the service for you.

– Fast turnaround
– Quality data
– Low cost

Please email or call 1-510-782-9988 x 2 to discuss your projects.