How to determine the DNA content of given sample by diphenylamine reaction.

When DNA is treated with diphenylamine acidic conditions a blue compound is formed with sharp absorption maxima at 595 nm. This reaction is given by 2’- deoxy pentose in general and is not specific for DNA. In acid soln the straight chain form of deoxy ribose is converted to highly reactive &-hydroxyl laevulin aldehyde which reacts with biphenyl amine to give blue columned complex. In DNA only the deoxyribose of Purina nucleotide reacts so that the value obtained represents half of the total deoxyribose present.

Requirement: – 1) std. DNA soln (1 mg/ml)
2) Diphenylamine reagent

Procedure: – To 1 ml of sample and std. add 2 ml water add 5 ml of diphenylamine reagent. Heat the soln on a water bath for 10 mins. and cool measure absorbance at 595 nm against water blank.

This website is a guide for for pharmacy students , pharmaceutical professional working in pharmaceutical regulatory affairs , pharmaceutical manufacturing , quality assurance , pharmaceutical formulation development , research and development.
This week we are publishing a series of articles on biochemistry, over this website , for our pharma students world wide .
We recomend you to sebscribe to this website and receive our articles directly by email when ever we update this website.

Following are some of articles which;will be useful for you in further understanding of aspects of sterile dosage form manufacturing

Pharmaceutical Validation

Cleen Room Classification

Pharmaceutical Industry

Pharmaceutical Manufacturing

Clinical Trials

Media fill

Regulatory Affairs in Pharmaceutical Industry

Leave a Reply

%d bloggers like this: