Title: Standard operating procedure of taking absorbance or transmittance reading on UV/VIS Spectrophotometer.
1.0 OBJECTIVE:
To lay down the procedure for analysis
on UV/VIS Spectrophotometer.
2.0 SCOPE:
This shall be applicable for analysis
on UV/VIS Spectrophotometer
of Quality Control Instrument Lab.
3.0 RESPONSIBILITY:
3.1
Technical Assistant.
3.2
Q.C Analyst.
4.0 ACCOUNTABILITY:
4.1
Manager-Quality Control.
4.2
Head of the Q.C Department.
5.0 MATERIAL AND EQUIPMENT:
5.1
UV/VIS Spectrophotometer.
5.2
Blank solution.
5.3
Sample solution.
6.0 PROCEDURE:
6.1
Ensure that the area is clean.
6.2
Switch on the main power.
6.3
Switch on the instrument.
6.4
Allow to stabilize for 15min.
6.5
Set the desired wavelength with knob in increasing order.
6.6
Keep the sensitivity switch at the highest position.
6.7
Put the mode selector at %T position.
6.8
Set the filter wheel as per the wavelength.
6.9
Adjust zero percent transmittance with the help of set zero
control.
6.10
Rinse reference cuvette and sample cuvette with blank
solution 2 to 3 times.
6.11
Adjust 100% T with the help of % knob.
6.12
Remove the cuvette and drain out the blank solution from it.
Rinse it with sample 2 to 3 times and filled with sample.
6.13
Clean the outer surface of the reference cuvette with tissue
paper.
6.14
Place the cuvette into the sample holder and cover the
cuvette compartment.
6.15
The reading show on the data position is the absorbance or
%T of the sample.
6.16
Record the results and make entry in instrument usage log
book.
6.17
Remove both the cuvette and wash with purified water or
methanol as required.
6.18
Calibrate the instrument by running similar way and make
entry in the calibration record.
6.19
Calibration frequency:
6.19.1
Monthly and after maintenance.
7.0 REVISION LOG:
Revision No. |
Effective Date |
Reason |
00 |
|
New SOP |
8.0 REFERENCES:
8.1
The British Pharmacopoeia. Vol V., Official Monograph / Ultraviolet and Visible Absorption
Spectrophotometry: 2015, Appendix: IIB
pp. 169-170.
9.0 ANNEXURES:
Calculations of UV/VIS
spectrophotometer.
Annexure:
1
Annexure:
2
Calculations of UV/VIS spectrophotometer
Annexure: 1
UV/VIS spectrophotometer Model:
_____________________________ Date:
_________________ OBSERVATIONS:
CALCULATIONS: Results:
_______________ Remarks:
_____________________________________________________________ |
Annexure: 2
UV/VIS spectrophotometer Model:
_____________________________ Date:
_________________ OBSERVATIONS:
CALCULATIONS: By
using formulas: 1.
1cmA1% = __a__ b.c 2.
%age purity= __Absorbance of
sample__ x 100 Absorbance
of standard 3.
Quantity of tablet = __stated
amount x %age purity.
100 Results:
_______________ Remarks:
____________________________________________________________ |