Confidentiality level: xxx

STANDARD OPERATING PROCEDURE

CODE XXX

UV-Vis Spectrophotometer Assay

Author: Accepted:
dd Mmm yyyy dd Mmm yyyy

Name of responsible analyst Name of approver

Responsible analyst Approver/Position
Pharmaceutical Testing Laboratory Pharmaceutical Testing Laboratory

Version Number XX
Supersedes version: XX
Dates of validity: dd Mmm yyyy –
dd Mmm yyyy

Name of Manager
Quality Manager

Distribution list:

Replaced by version: XX
Standard Operating Procedure Version No XX
CODE XXX UV-Vis Spectrophotometer Assay Dd Mmm yyyy Confi Level XXX

1. Objective

2. Responsibility
1. The person assigned to be in charge of this instrument should follow the instructions in
this procedure. Any changes should be reported to the head of the laboratory. Proper
revisions should be done immediately upon approval of the head of the laboratory.
2. The person assigned to be in charge of this instrument should train staff and students
on the operation of this equipment and to ensure adherence to the working instruction.
3. The person assigned to be in charge of this instrument is identified on the logbook
cover pages for this instrument.

3. Definitions

Acceptance criteria Measurable terms under which a test result will be considered
acceptable. [2]

Calibration One of the primary processes used to maintain instrument

accuracy; it is a determination of the deviation between
measurements and the true value under specified measuring
conditions. It is the process of configuring an instrument to provide
a result for a sample within an acceptable range. [2]

Drug Any substance or pharmaceutical product for human or veterinary

use that is intended to modify or explore physiological systems or
pathological states for the benefit of the recipient [2]

Reference standard USP Reference Standards are highly characterized specimens of

drug substances, excipients, reportable impurities, degradation
products, compendial reagents, and performance calibrators. They
are explicitly required in many Pharmacopeial assays and tests and
are provided solely for such use. [3]

4. References

 FDA Circular No. 2013-014 [1]

 WHO TRS 937 (2006) [2]

 USP 35-NF 30 [3]

5. Procedures

  Steps
SAMPLE PREPARATION
Standard Operating Procedure Version No XX
CODE XXX UV-Vis Spectrophotometer Assay Dd Mmm yyyy Confi Level XXX

1 Filter the solution if necessary.

2 Dilute the filtrate with the suitable medium as indicated in the individual monograph.
Transfer the premixed solution in a suitable cuvette.
3
Note: Clean the cuvette before and after use. Wash the cuvette using the solvent that is to
be used in the experiment.
Compare the test solution with a standard solution having a known concentration of USP
4
Reference Standard in the same medium, if necessary.
POWERING UP
Press the power button to turn on the instrument. The display should illuminate. The
1 table indicating the power up diagnostics of the parameters regarding the basic
functions of the equipment will be displayed.
Flush the flow cell with cleaning solution and distilled water and turn off the instrument
2
in case any parameters fail.
3 Continue the process until the instrument successfully completes power up diagnosis.
4 Exit the power up diagnostics upon completion of the process.
CALIBRATION
1 Start calibration by clicking the appropriate button in the menu bar.
Allow the instrument and lamp source to warm up and stabilize for approximately 30
2
minutes.
3 Select the wavelength mode.
Prepare calibration blank using distilled water or extracting solution and all reagents that
4
will be used in the samples according to the proper procedure.
5 Change instrument mode to absorbance mode “Abs mode”
Click on the field to the right of the “λ” and input the proper wavelength for the analyte
6
being tested.
Draw the blank into the flow cell by inserting the sipper tube into the culture tube and
7
pressing the fill button on the pump.
Verify the proper connection between the instrument and all output devices prior to
8
performing any analytical operations.
9 Start sample analysis once all connections have been checked.
OPERATION
1 Power up instrument as described above.
2 Calibrate the instrument as stated above.
3 Set the analytical order in the following manner: blank solution, standard and samples.
4 Verify the initial calibration and initial blank at the beginning of an analytical run.
UV-VIS PROCEDURE
Turn on the UV-Vis spectrophotometer by pressing the power button. This unit is not
1
operational until the blinking light on the button goes to a solid green color.
2 Turn on the computer and login.
3 Open the lid of the sample compartment.
Standard Operating Procedure Version No XX
CODE XXX UV-Vis Spectrophotometer Assay Dd Mmm yyyy Confi Level XXX

Check inside the UV-Vis chamber to assure that the appropriate sample holder is in
4
place.
5 Place the cuvette containing the prepared samples in the cell holder.
6 Close the lid of the sample compartment
7 Set the experimental parameters as stated in the individual monograph.
8 Click scan to start the program.
9 Save the results in the system.
10 Record the results in the analyst workbook.
11 Open the lid and remove the cuvettes from sample cell holder.
12 Close the lid and turn off the equipment.
13 Wash the cuvettes used with tap water and dry it using lint-free tissue paper.
14 Store the cuvettes properly.

6. Distribution and Retrieval

DISTRIBUTION RETRIEVAL
CONTACT
NAME No. SIGNATURE DATE SIGNATURE DATE
           
           
           
           
           
           
           
           
           
           

7. Revision History

Date/Initials* Version Section, page: Change / Reason to change** New version

Dd Mmm yyyy/XXX
Standard Operating Procedure Version No XX
CODE XXX UV-Vis Spectrophotometer Assay Dd Mmm yyyy Confi Level XXX

*Initials of the person making the change

**When not self-evident