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Study Notes Quantification Methods

The document outlines quantification methods in biochemistry, focusing on the anthrone method for estimating carbohydrates and the ninhydrin method for amino acids. The anthrone method involves dehydrating carbohydrates to form compounds that react with anthrone reagent, while the ninhydrin method produces colored complexes with amino acids based on their functional groups. Both methods emphasize the importance of standard curves and blank solutions for accurate measurements.

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3 views2 pages

Study Notes Quantification Methods

The document outlines quantification methods in biochemistry, focusing on the anthrone method for estimating carbohydrates and the ninhydrin method for amino acids. The anthrone method involves dehydrating carbohydrates to form compounds that react with anthrone reagent, while the ninhydrin method produces colored complexes with amino acids based on their functional groups. Both methods emphasize the importance of standard curves and blank solutions for accurate measurements.

Uploaded by

moengk16
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Study Notes: Quantification Methods in Biochemistry

1. Quantification of Carbohydrates Using the Anthrone Method

Date: 23 April 2025

Topic: Carbohydrates Estimation

Principle:

- The anthrone method is a colorimetric technique used to estimate the amount of carbohydrates in a sample.

- Carbohydrates are dehydrated by concentrated sulfuric acid to form furfural (from pentoses) or

hydroxymethylfurfural (from hexoses).

- These compounds react with anthrone reagent to produce a green-blue complex, which is measured at 620

nm using a spectrophotometer.

Purpose of Glucose Standard Curve:

- To create a reference plot of absorbance vs. glucose concentration.

- Helps in determining the unknown concentration of carbohydrates in test samples.

- Ensures accuracy and reliability of the measurement.

2. Quantification of Amino Acids Using the Ninhydrin Method

Date: 28 March 2025

Topic: Amino Acids Estimation

Principle:

- Ninhydrin reacts with free amino groups in amino acids to form a colored complex.

- Most amino acids give a purple color (Ruhemann's purple).

- Amino acids like proline and hydroxyproline produce a yellow color due to their secondary amine groups.
Study Notes: Quantification Methods in Biochemistry

- The intensity of the color is measured spectrophotometrically to determine amino acid concentration.

Amino Acids Reacting with Ninhydrin:

- All amino acids with free primary amine groups produce Ruhemann's purple.

- Proline and hydroxyproline yield yellow due to their imino groups.

Purpose of Blank Solution:

- Serves as a reference to eliminate absorbance due to solvents and reagents.

- Contains everything except the test sample (i.e., solvent + reagent only).

- Used to zero the spectrophotometer for accurate readings.

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