1/31/2014

Calibration of UV / Visible Spectrophotometer : Pharmaceutical Guidelines

Calibration of UV / Visible Spectrophotometer

Learn how to calibrate the UV - Visible spectrophotometer using Potassium dichromate Solution. Ensure that the connections of the instrument are proper. Control of Absorbance Potassium dichromate Solution : Dry a quantity of potassium dichromate by heating to constant weight at 130C. Weigh & transfer accurately a quantity not less than 57.0 mg & not more than 63.0 mg to 1000 ml volumetric flask. Dissolve & dilute in sufficient 0.005M H2SO 4 to produce 1000 ml. Measure the absorbance of potassium dichromate solution at the wavelengths given below. Calculate value of A (1% 1cm) for each wavelength. A (1% 1cm) = Absorbance X 10000 / Weight of Potassium dichromate in mg. Acceptance criteria Wavelengths (nm) A [1% 1cm] Limit 235.0 124.5 122.9 - 126.2 257.0 144.0 142.8 - 145.7 313.0 48.6 47.0 - 50.3 350.0 106.6 105.6 - 108.2 Stringent limits are adopted from IP & BP for acceptance criteria Resolution power Record the spectrum of a 0.02% v/v solution of toluene in hexane in the range of 260 nm to 420 nm (before use check the hexane for transmittance, using water as a blank between 260nm to 420nm & use only if transmittance is not less than 97%). Acceptance criteria: The ratio of the absorbance at the maximum at about 269nm to that at the minimum at about 266nm is not less than 1.5. Limit of Stray Light Prepare a 1.2 % w/v solution of Potassium chloride in water. Measure absorbance of the above solution at 198.0, 199.0, 200.0, 201.0, 202.0 nm using water as blank. Acceptance criteria: Absorbance is greater than 2. Wavelength accuracy, Resolution & Baseline flatness (inbuilt test) Attach printer directly to the instrument (instead of computer). Go to MODE press F3 key i.e. Maintenance. Press 1. Press Start/Stop key. After screen changes, ensure that nothing is kept in the optical path & press the Start/Stop key again. Prints will come after all the three tests are over. Acceptance criteria: For wave length accuracy : at 656.1 0.3 nm & at 486.0 0.3 nm. Resolution: 1.0 nm or less. Baseline flatness: 0.002Abs. Frequency: Once in a month. Maintenance / Repairs When the instrument does not comply with the requirement / tolerance range specified above, the instrument should be labeled OUT OF CALIBRATION and should be repaired / serviced. After repair / maintenance, calibrate the instrument. REFERENCE Indian Pharmacopoeia British Pharmacopoeia Instruction manual of UV-1700.

CALIBRATION REPORT OF UV / VISIBLE SPECTROPHOTOMETER

Page 1 of 2 SOP Reference Date Instrument No. Frequency : Once in a month Model Make 1. Control of Absorbance: Potassium dichromate solution : Gross Wt. : . Balance : Tare Wt. : . Net Wt. : . .. ml with 0.005M sulphuric acid. Dissolve and dilute 57.0 to 63.0 mg of potassium dichromate in sufficient 0.005M sulphuric acid to produce 1000.0 ml. Wavelength (nm) Absorbance A (1%, 1cm) Tolerance

http://www.pharmaguideline.com/2010/05/calibration-of-uv-visible.html

1/3

1/31/2014
235.0 257.0 313.0 350.0 Calculation :
1%

Calibration of UV / Visible Spectrophotometer : Pharmaceutical Guidelines

122.9 142.8 47.0 105.6 Absorbance x 10000 = ------------------------------------------------------to 126.2 to 145.7 to 50.3 to 108.2

A1 cm

Weight of potassium dichromate in mg Remark : Satisfactory / Not satisfactory. Performed by : Checked by : Date : Date : 2. Resolution power : 0.02% v/v toluene in hexane : .ml 100.0 ml with hexane / ml 100.0 ml with hexane Absorbance at 269 nm = Absorbance at 266 nm = Absorbance at 269 nm Ratio = ---------------------------------- = ---------------- = Absorbance at 266 nm (Limit : Not less than 1.5) Remark : Satisfactory / Not satisfactory. Performed by : Checked by: Date : Date :

CALIBRATION REPORT OF UV / VISIBLE SPECTROPHOTOMETER

3.

Page 2 of 2 Limit of stray light: Potassium chloride solution Gross Wt. : . Balance : Tare Wt. : . Net Wt. : . . ml with water. Dissolve and dilute accurately 1.2 g potassium chloride in sufficient water to produce 100.0 ml. Sr. No. Wavelength (nm) Absorbance Tolerance 1 198.0 2 199.0 Absorbance is greater 3 200.0 than 2 4 201.0 5 202.0 Remark : Satisfactory / Not satisfactory. Performed by : Checked by : Date : Date : 4. Wavelength accuracy (In built test) Sr. No. 1 2 Wavelength Standard (nm) 656.1 486.0 Checked by: Date : Observation nm Checked by: Date : Observation Wavelength Observed (nm) Difference Limit (nm) + 0.3 + 0.3

5.

6.

Remark : Satisfactory / Not satisfactory. Performed by : Date : Resolution (In built test) Limit Sr. No. 1 1.0 nm or less Remark : Satisfactory / Not satisfactory. Performed by : Date : Baseline flatness (In built test) Limit Sr. No. 1 0.002 Absorbance Remark : Satisfactory / Not satisfactory. Performed by : Date :

Checked by: Date :

Conclusion : Instrument working is Satisfactory / Not satisfactory. Next calibration due on : Performed by : Date : Checked by : Date :

http://www.pharmaguideline.com/2010/05/calibration-of-uv-visible.html

2/3

1/31/2014

Calibration of UV / Visible Spectrophotometer : Pharmaceutical Guidelines

CGMP ISO USFDA WHO FOLLOW CONTACT US PHARMA WIDGETS

2013 All rights reserved to Ankur Choudhary and data may not be reproduced on other websites. Designed by Pharmaceutical Guidelines.

http://www.pharmaguideline.com/2010/05/calibration-of-uv-visible.html

3/3