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C are the absorbances of the sample and standard solutions respectively.
It involves measuring the amount of ultraviolet or visible radiation absorbed by a substance in solution.
Instrument which measure the ratio, or function of ratio, of the intensity of two beams of light in the U.
The fundamental law that governs the quantitative spectrophotometric analysis is the Beer -Lambert law.
Beer’s law: It states that the intensity of a beam of parallel monochromatic radiation decreases exponentially with the number of absorbing molecules.
The wavelength normally selected is wavelength of maximum absorption (λ), where small error in setting the wavelength scale has little effect on measured absorbance.
Ideally, concentration should be adjusted to give an absorbance of approximately 0.9, around which the accuracy and precision of the measurements are optimal.
In other words, absorbance is proportional to the concentration.
Lambert’s law: It states that the intensity of a beam of parallel monochromatic radiation decreases exponentially as it passes through a medium of homogeneous thickness.
In calibration graph method, the absorbances of a number of standard solutions of the reference substance at concentrations encompassing the sample concentrations are measured and a calibration graph is constructed.
The concentration of the analyte in the sample solution is read from the graph as the concentration corresponding to the absorbance of the solution.
Spectrophotometric; Developed; Validated; Parameters; ICH[Q2(R1)] Spectroscopy methods [1,2] It is the branch of science dealing with the study of interaction between Electromagnetic radiation and matter.