Ad Code

Absorbance Reading on UV/VIS Spectrophotometer.

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:

Thickness of cell:

 

Spectral range:

 

Maxima absorption wavelength

 

Sample:

 

Other reagent used:

 

 

No. of obs.

Concentration

Wavelength

Absorbance

 

 

 

 

 

 

 

 

 

CALCULATIONS:

 

 

 

 

 

Results: _______________

Remarks: _____________________________________________________________

 

 

Annexure: 2

UV/VIS spectrophotometer

Model: _____________________________                             Date: _________________

OBSERVATIONS:

No. of tablets:

 

Sample:

 

Thickness of cell (b):

 

Concentration solution (c):

 

At A(1%, 1cm):

 

Wavelength (λ):

 

 

No. of obs.

Concentration

(c)

Wavelength

(λ)

Absorbance

(a)

Specific absorbance

A (1%,1cm)

 

 

 

 

 

 

 

 

 

 

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: ____________________________________________________________