SOP for Operation and Calibration of Polarimeter.

1.0        Objective:

                  The purpose of this SOP is to provide written procedure for Operation and Calibration       of Polarimeter.

2.0      Scope:

           This SOP covers Operation and Calibration of Polarimeter.

3.0           Responsibility:

       Jr. Research Officer, Research Officer: Responsible for operation, calibration and maintenance of the instrument as per procedure.

     Head of Department: Responsible for calibration and maintenance, timely as per   schedule.

         QA Officer/QA Manager: Review the records and governing the document.

4.0             Procedure:

4.1             Ensure that the instrument is visibly clean and free from dust & all the parts including optical lens, telescope lens, Polarimeter tube are clean.           

4.2      Set up the instrument on the plain platform with the analyzer head facing the operator, and illuminated sodium lamp attached to the bracket on the instrument in direct line with the axis of the instrument and about four inches from the end.

4.3        Turn the power on.

4.4      Wait for 20 minutes to elute the sodium lamp to its maximum brightness.

4.5         Focus the top eyepiece to show the Scale and index line.

4.6        Remove the Polarimeter tube or keep the tube empty.

4.7      Now set the eye of the field telescope and focus to obtain a circular disc of light.

4.8             By means of wheel type knob from side arm in such a position that the light field is  divided into two halves rapidly changing in light intensity as the control knob is moved and that between the extremes of blackout of the two halves, such that a position may be found that gives equal black intensity.

4.9             This is the balance position or zero - zero position (The position that if you slightly turn the know left side half black shade will seen and if you turn slightly back the right side half black shade will seen). Here, the half-dark and half-light portion is matched with a knob by rotating it in clockwise or anticlockwise directions. It is also defined as dextro ( + ) or leavo ( -  ) rotatory respectively.

4.10  At the balance position circle should read zero and micrometer is set to zero.

4.11    Now clean the Polarimeter tube and window glass pieces with distilled water.

4.12        Rinse the Polarimeter tube with the media.

4.13    Close the one end of the tube, fill the tube with   media (solution of an optically active sample whose activity is to be measure) and slide it down in the polarizing unit. Be sure that there is no air bubble large enough to block the light path inside the polarimeter tube.

4.14       By means of the control wheel knob readjust to obtain the same balance position as in 4.8.  Record the reading as a blank reading.

4.15     Repeat the above step 4.14 and 4.15 for test sample. Take at least five readings for test sample. Record the mean of these five reading as a test reading (i.e. angle of optical rotation).

4.16  The difference between the readings obtains with and without the sample being the measure is the sample optical activity.

4.17       Clean the instrument after use.

4.18      Enter the details of the sample in the Polarimeter usage log book (Annexure – 1).

5.0        Calibration:

5.1    Calibrate the instrument by using different concentrations of Sucrose previously dried at 105° C for 2 hours.    

             

CONCENTRATION             ANGLE OF ROTATION      TOLERANCE

              (g /100 ml)     at  25° C             in degree (+)

                  10.0                                               13.33                                       ± 0.05

                  20.0                                               26.61                                       ± 0.05

                  30.0                                               39.86                                       ± 0.05

                 40.0                                                53.06                                       ± 0.05

                 50.0                                                66.23                                       ± 0.05

5.2          Record the observations in the polarimeter calibration format as given in Annexure - 2.

5.3          Calibration frequency: Once in a month and after each maintenance job.

6.0          Routine maintenance:

6.1        Clean the instrument with a clean dry cloth.

6.2        Clean the eye piece with tissue paper.

7.0          Documentation:
7.1    Annexure - 1   -Polarimeter usage log book.           XXX/FRM/000
7.2    Annexure - 2   -Polarimeter calibration Format.     XXX/FRM/000

8.0         History of Revision:

Revision No.

Effective Date

Revision details

Reason for revision

01

DD/MM/YEAR

Add revision history, routine maintenance and defined responsibility.

Periodic review

Annexure – 1
Date	Name of Sample	A. R. No.	Batch No.	Used by	Remark
SOP No. : XXX/SOP/000-00                                                         Format No. : XXX/FRM/000-00

Annexure – 2

Calibration of Polarimeter Calibrated Date   :                                                         Next due date for calibration : Make               :                                                             Model No. :

Calibration : Sucrose Make  :                                                            Batch No.  :   Preparation of Solution ( 50 % w/v) : à Take 50.0 gm of sucrose and dissolved in water to make 100 ml with Purified water. (Solution-A; 50 % w/v) à Take 40 ml of solution-A in 50 ml volumetric flask and make volume with Purified water. (Solution-B; 40 % w/v) à Take 30 ml of solution-A in 50 ml volumetric flask and make volume with Purified water. (Solution-C; 30 % w/v) à Take 25 ml of solution-B in 50 ml volumetric flask and make volume with Purified water. (Solution-D; 20 % w/v) à Take 25 ml of solution-C in 50 ml volumetric flask and make volume with Purified water. (Solution-E; 10 % w/v) Sr. No. Concentration of Sucrose (%) w/v Angle of Rotation (+) Observed at 25° Limits of Angle of Rotation (+) at 25° 01 10.0 13.28 – 13.38 02 20.0 26.56 – 26.66 03 30.0 39.81 – 39.91 04 40.0 53.01 – 53.11 05 50.0 66.18 – 66.28

Remarks : Satisfactory / Not Satisfactory

Calibrated by  :                                                                      Checked by      :

SOP No. : XXX/SOP/000-00                                                   Format No. : XXX/FRM/000-00