Richard hypothesis 

            

TLS Online TPP Program

#Id: 4723

#Unit 13. Methods in Biology

The absorption spectrum of a chromophore is only partly determined by its chemical structure. The environment also affects the observed spectrum, which mainly can

be described by three parameters:

protonation/deprotonation (pH, RedOx);

• solvent polarity (dielectric constant of the solvent); and

• orientation effects.

TLS Online TPP Program

#Id: 4724

#Unit 13. Methods in Biology

Concentration of protein can be analyzed by to method constructive and destructive method 

TLS Online TPP Program

#Id: 4725

#Unit 13. Methods in Biology

Destructive method uses Folin reagent, Biuret, BCA, Coomassie Brilliant Blue and give absorbance at 660, 540, 562, 595 nm of wavelength respectively.

TLS Online TPP Program

#Id: 4726

#Unit 13. Methods in Biology

The constructive method involved direct analysis of protein and gives absorbance at wavelength of 280 nm.

TLS Online TPP Program

#Id: 4727

#Unit 13. Methods in Biology

Carotenoids gives absorbance between 400 – 500 nm of wavelength while porphyrin gives absorbance at 400 nm of wavelength known as Soret band.

TLS Online TPP Program

#Id: 4728

#Unit 13. Methods in Biology

DNA gives absorbance at 595 nm in presence of Diphenylamine while gives direct absorbance at 260 nm.