Procedure for Operation and Calibration of Photofluorimeter

This document details the procedure for the operation and calibration of the Photofluorimeter. This SOP is applicable at the time of the operation and calibration of the Photofluorimeter.

It is the policy of XYZ Pharmaceuticals Limited that a written procedure shall be followed for the operation and calibration of the Photofluorimeter in order to ensure smooth operation and its use monitored to obtain consistent and reproducible results and minimize downtime and meet regulatory requirements.


Persons along with their responsibilities and accountability are given below:

Executive  - Corporate Quality Assurance:- To prepare the SOP

Trainee Analyst/ Jr. Analyst/ Analyst/ Sr. Analyst:- To follow the SOP accordingly

Vice President – Technical:- To ensure implementation of SOP.


CLEANING Procedure for TUBE

Never use a brush to clean the tube of the Photofluorimeter. Clean the tube using the purified water or any organic solvent (i.e. acetone or methanol). Add about 1 ml of the solution to be measured. Tilt and turn the tube so that the solution has contact with the entire surface. Discard the solution and repeat this rinse once more.

5.1.4 Wipe the outside of the tube with a lint-free, soft tissue to remove any moisture or fingerprints from the outside surface

5.2 PRECAUTIONS for Photofluorimeter

5.2.1   Allow the instrument to warm up for a period of 30 minutes prior to use.

5.2.2   Switch “ON” the instrument and see if the neon lamp on the front panel is glowing or not.

5.2.3    Check if the cooling fan is running or not.

5.2.4    Avoid handling the lower part of the sample tube.

5.2.5    Clean the primary and secondary filter properly with the lint-free cloth.

5.2.6    Each measurement requires a minimum of 4 ml of sample solution.

Test tubes must be inserted in the sample holder with the index mark (or trademark) directly facing the operator

5.3 OPERATION Photofluorimeter


5.3.2 Set the “standard coarse” control and the “blank coarse” control to a minimum i.e. counterclockwise.

5.3.3 Insert appropriate “primary” and “secondary” filters on their respective slots.

5.3.4 Switch on the instrument. Check if the cooling fan is running this can be noticed by slight noise from the fan and the flow of air from the perforated sheet at the back.

5.3.5 Keep the instrument “ON” and allow a warm-up period of 30 minutes prior to use.

5.3.6 Wash, rinse and fill the test tube with a blank solution.

5.3.7 Keep the test tube in the sample holder of the instrument.

5.3.8 Match the test tube mark with the mark in the sample holder.

5.3.9 Close the shutter.

5.3.10 Push the button marked “TO READ” and set the pointer approximately at “ZERO”

5.3.11 Insert the tube containing the standard solution of known concentration into the sample holder.

5.3.12 Push the button “TO READ” and set the meter reading to 50 (or as per concentration) using the standard Knob.

5.3.13 Take the readings in triplicate repeating the above procedure.

5.3.14 Replace the standard solution with the blank solution and note the meter reading.

5.3.15 Replace the blank solution with the unknown solution (sample solution) and note   the reading in triplicate repeating the above procedure

5.3.16 Blank reading must be subtracted from the standard and the unknown sample before comparison.

5.3.17 Switch off the instrument after use

5.3.18 Calculate the concentration, Cx of the substance in the solution to be examined, using the expression.    C  =    Ix   X   Cs 


Where   Cx   = Concentration of the solution to be examined

Cs  = Concentration of the standard solution

Ix   = Intensity of the light emitted by the solution to be examined

Is    = Intensity of the light emitted by the standard solution                                                         

5.3.19 Maintain Logbook of Photoflourimeter as per XYZ/CQA/SOP-063/FR-01, Logbook for Photofluorimeter Appendix I.

Procedure for Operation and Calibration of Photofluorimeter

6.0  CALIBRATION of Photofluorimeter

6.0.1 Calibration Frequency: Quarterly

6.0.2 Before attempting calibration the photofluorimeter must be switched “ON” for at least 20 minutes.

6.0.3 Weigh accurately about 50 mg of Quinine Sulphate and dilute it up to 500 ml with 0.05M Sulphuric acid (Solution A).

From Solution A prepare the solution of different concentrations in 0.05 M sulphuric acid as per the following table


Vol. of Solution A taken


Makeup to (mL)

Final concentration (ppm)


























Set the instrument to read 50 on the scale with 8ppm (solution G) Quinine Sulphate solution.

Take reading in triplicate and record the calibration data on XYZ/CQA/SOP-063/FR-02 as Calibration Datasheet Appendix II (page 1of 1).

Similarly, record the reading in triplicate for the remaining solutions (from solution B to F) of Quinine Sulphate solution.

Plot a graph of concentration v/s average of the readings taken. The graph should be linear and the coefficient of correlation should be close to 1.

After the calibration of the Photofluorimeter affix a calibration sticker on the instrument as per XYZ/CQA/SOP-006, Labelling policy, and fill in all the required details.

Ensure the power supply to the instrument is switched “OFF” before clearing. Clean the instrument with a clean dry swab every day.

6.0.11 Report any discrepancy observed during operation and calibration of instrument to. To Section In-charge or his representative for corrective and preventive action.

6.0.12 Section In-charge or his representative will take the necessary action and report the same to Manager – Quality Assurance.

6.0.13  Affix ‘Out of Order’ label on Photoflurimeter.


Generate the instrument calibration number on the calibration datasheet as INSCALXXYYZZZ where INS denotes instrument, CAL denotes calibration, XX denotes the year, YY denotes month and ZZZ denotes sequence number.


XYZ                 XYZ Pharmaceuticals Limited

CQA                Corporate Quality Assurance.

QA                   Quality Assurance

SOP                Standard Operating Procedure

>Operation and Calibration of the Polarimeter

LogBook And Appendix:- Download

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