INSTRUMENT CALIBRATION

UV
 CONTROL OF WAVELENGTHS:

– Use 4% solution of Holmium oxide in 1.4M Perchloric acid

– Use UV-Visible standard Holmium oxide filter/cuvette
– Scan Holmium oxide solution/cuvette in range of 200 nm to 600 nm.
– Record the spectrum and the maxima observed.

– Acceptance criteria:
Tolerance limit for
Actual Wavelength
Wavelength
241.15 nm ± 1 nm
287.15 nm ± 1 nm
361.50 nm ± 1 nm
536.30 nm ± 3 nm
 CONTROL OF ABSORBANCE:

Prepare 60ppm solution of dried potassium dichromate in 0.005 M sulfuric

acid solution.

Check the absorbance at wavelengths of 235 nm, 257 nm, 313 nm and 350
nm using 0.005 M sulphuric acid as blank solution.
Calculate Specific absorbance, A (1%, 1 cm) using the formula:
 Acceptance criteria
Wavelength (nm) A (1%, 1 cm ) Acceptance Criteria

235 124.5 122.9 to 126.2

257 144.5 142.8 to 146.2

313 48.6 47.0 to 50.3

350 107.3 105.6 to 109.0

430 15.9 15.7 to 16.1

 LIMIT OF STRAY LIGHT:

• Prepare 12 gm/L Potassium chloride solution.

• Scan the 12 g/L Potassium chloride solution in the range of

195 nm to 220 nm, record the spectrum and determine the
maximum absorbance value between 198 nm and 202 nm.

• Acceptance Criteria: The absorbance in a 1 cm cell increases

steeply between 220 nm and 200 nm and is greater than 2.0
at 198 nm.
 RESOLUTION POWER:

Prepare 0.02 % v/v Toluene in n-Hexane

Note down the absorbance and determine the ratio of absorbance at the maxima
and minima at about 269 nm and 266 nm respectively.

Absorbance at maxima at about 269 nm

Ratio = _____________________________________________________

Absorbance at minima at about 266 nm

Acceptance Criteria:

The ratio of the absorbance at the maxima and minima at about 269 nm and 266
nm respectively should be not less than 1.50.