Instrument Calibration Protocol for UV-Visible Spectrophotometer

INSTRUMENT	UV-Visible Spectrophotometer
Instrument ID. No.
Ref. SOP No.
Model No.	Perkin
Calibration frequency
Date of calibration
Next due date

1. Control of Absorbance :

  Balance ID : _________________

  Potassium Dichromate Lot No. / Batch No. : ___________________

 Potassium Dichromate Expiry / Valid Up to : ___________________

For 235 nm, 257 nm, 313 nm and 350 nm :

Weight of Potassium Dichromate: ___________ g.

For 430 nm :

Weight of Potassium Dichromate: ___________ g.

Wavelength (nm)	Absorbance	A (1%, 1 cm)	STD A (1 %, 1 cm)	Limit	Remarks
235			124.5	122.9 to 126.2
257			144.0	142.8 to 146.2
313			48.6	47.0 to 50.3
350			106.6	105.6 to 109.0
430			15.9	15.7 to 16.1

Where,

A (%, 1cm) = [A / (C x D)]

A – Observed Absorbance

C – Concentration of Potassium dichromate solution in % as w/v.

D – Path Length in cm.

2. Resolution :

Spectroscopic Hexane Lot No. / Batch No. : ___________________

Spectroscopic Hexane Expiry / Valid Up to : ___________________

Spectroscopic Toluene Lot No. / Batch No. : ___________________

Spectroscopic Toluene Expiry / Valid Up to : ___________________

Observation :   

Absorbance at about 269 nm : _________

Absorbance at about 266 nm : _________

Ratio : = Maximum Absorbance/ Minimum Absorbance =

Limit: Not Less than 1.5.

3. Limit of Stray Light :

Balance ID : _________________

Potassium Chloride Lot No. / Batch No. : ___________________

Potassium Chloride Expiry / Valid Up to : ___________________

Weight of Potassium Chloride  : ___________ g.

Observation :

Absorbance at 200 nm : _________

Limit   : Not Less than 2.0.            

4. Baseline Flatness :

Observation : Observed Baseline Flatness : _________

Limit   :  Baseline Flatness shall be within ± 0.002 Abs.

5. Second derivative Spectrophotometer : Resolution Power :

 Spectroscopic Toluene Lot No. / Batch No.  : ___________________

Spectroscopic Toluene Expiry / Valid Up to : ___________________

Methanol AR Grade Lot No. / Batch No.      : ___________________

Methanol AR Grade Expiry / Valid Up to     : ___________________

 Observation :

Ratio : A/ B =

Limit   : Not less than 0.2.

6. Control of Wavelength:         

Balance ID: _________________

Holmium Oxide AR Grade Lot No. / Batch No. : ___________________

Holmium Oxide AR Grade Expiry / Valid Up to : ___________________

Weight of Holmium Oxide AR: ___________ g.          

Wavelength (nm)	Observed Wavelength (nm)	Maximum Tolerance	Remarks
241.15		240.15 to 242.15
287.15		286.15 to 288.15
361.50		360.50 to 362.50
536.30		533.30 to 539.30

7. Control of Wavelength (In House Method) :

Balance ID : _________________

Caffeine AR Grade Lot No. / Batch No. : ___________________

Caffeine AR Grade Expiry / Valid Up to : ___________________

Weight of Caffeine AR   : ___________ mg.

Wavelength (nm)	Observed Wavelength (nm)	Maximum Tolerance	Remarks
205		204.0 – 206.0
273		272.0 – 274.0

Remarks: Calibration Satisfactory / Not Satisfactory

Calibrated By (Sign/ Date)

Checked By (Sign/ Date)