Factors Affecting Absorption Spectra

• Physical Factors
– PH
– Temperature
– Solvent Selection
– Concentration
– and Path length
• Chemical Factor
• Instrumental Factors
– Slit width
– Scan speed
– Stray light
 PHYSICAL FACTORS
PH

• By changing PH spectra is changed because ionization is

changed.
• If we need to check Benzoic acid concentration then we check
it in molecular form not in ionizable form.
• So we add Buffer to maintain the PH.
• Drug remain in molecular form.
 Temperature

• Evaporation of solvent and its conc. can be

change by changing temperature.
• At high temp. evaporation increase and
solution became concentrated.
• At high conc. Beer and Lambert's law is
deviated
• Absorbance is taken in A.C environment.
 Solvent Selection
• Compound should be soluble in solvent.
• Solvent should be transparent.
• Solvent must not show its own absorbance
where the sample is showing absorbance
• The absorbance of sample lies in the range of
0.5-1.5.