Estimation of RNA by Orcinol Method using single standard
Orcinol, furfural, RNA, theory of orcinol method This is a general method for the estimation of pentoses. Acid hydrolysis of RNA releases ribose which is the presence of strong acid undergo dehydration to yield furfural. Orcinol in the presence of ferric chloride as a catalyst, reacts with furfural producing a green coloured compound with absorbance maxima at 640nm. RNA gives a limited positive reaction with orcinol test. PROCEDURE • Test tubes were labelled as S, T and B for standard, test and blank respectively. • In tube labelled S, 1ml of standard solution was added. • 1.0 of test solution was added into test tube labelled ‘T’ • 2 ml of 0.1 N NaOH was added into tube labelled ‘B’ and all the tubes were made up to 2 ml with 0.1 N NaOH. • 2 ml of orcinol reagent was added to all tubes and kept the tubes in boiling waterbath for 30 minutes. • After cooling the absorbance of the solution was measured colorimetrically at 640 nm. The amount of RNA in the given unknown sample was calculated.
Orcinol, furfural, RNA, theory of orcinol method This is a general method for the estimation of pentoses. Acid hydrolysis of RNA releases ribose which is the presence of strong acid undergo dehydration to yield furfural. Orcinol in the presence of ferric chloride as a catalyst, reacts with furfural producing a green coloured compound with absorbance maxima at 640nm. RNA gives a limited positive reaction with orcinol test. PROCEDURE • Test tubes were labelled as S, T and B for standard, test and blank respectively. • In tube labelled S, 1ml of standard solution was added. • 1.0 of test solution was added into test tube labelled ‘T’ • 2 ml of 0.1 N NaOH was added into tube labelled ‘B’ and all the tubes were made up to 2 ml with 0.1 N NaOH. • 2 ml of orcinol reagent was added to all tubes and kept the tubes in boiling waterbath for 30 minutes. • After cooling the absorbance of the solution was measured colorimetrically at 640 nm. The amount of RNA in the given unknown sample was calculated.