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)