STANDARD OPERATING PROCEDURE LAB EQUIPMENT H.P.L.C

STANDARD OPERATING PROCEDURE

SUBJECT :                     LAB EQUIPMENT  H.P.L.C

DEPARTMENT           : QA SOP NO.                      : Page No                       : Effective Date             : Next Review                : SOP REVISION           :

1.0       OBJECTIVE:

To provide guidelines for Operation & Maintenance of H PLC System.

2.0       POLICY: 

To ensure establish a procedure for operation of H.P.L.C.

3.0       PROCEDURE:

3.1       Switch on the Power supply and UPS.

3.2       Switch on the Pump, Detector, computer and Printer, allow to warm the system.       

3.3       Check the Column fitted on the system is suitable or not, if not install the required Column.

3.4       Wash the Column with washing solution (methanol & water each) for 30 minutes.

3.5       Prepare the Mobile phase and degas carefully.

3.6       Run the Mobile phase at required flow rate to saturate the Column and check the baseline.

3.7       Set the required detector parameters on Detector.

3.8       If the base line is o.k. Then set the required chromatogram parameters on Method file or run the injections on default method first and set suitable parameters.

3.9       Prepare the standard solutions and run five standard injections on the system and check system suitability. The injecting sample does not have any air bubbles prior to injection.

3.10     If the RSD of the Standards chromatograms is less than two, then the system is suitable for Sample           injections. Otherwise repeat the Standard injections till you get the RSD below two.

3.11     Inject the required Sample and take the chromatograms and calculate the Percentage of Active ingredient on the basis of respective peak areas.

3.12     After completion of work, the Column must be washed with wash solution for one hour

3.13     For better performance of HPLC system adhere to the following guidelines:

3.13.1  Use only HPLC grade chemicals.

3.13.2  Mobile phase and all solutions of HPLC must be filtered and degassed.

3.13.3  Filter the standard and sample solutions using 0.45 micron nylon 66 membrane filter.

3.13.4  Do not operate the system above 3000psi.

4.0       FREQUENCY:  As and when required.

5.0       RESPONSIBILITY:  Executive Q.A.

6.0       REVIEW GUIDE: Log register & Calibration register.

7.0       ANNEXURE: -------

8.0       SOPs DISTRIBUTION:

            This SOP shall be distributed to following Departments:

Name of the Department	Master Copy	Circulatory Copy
QA/QC	√	√
Production	x	X
Warehouse	x	X
General	x	X
Maintenance	x	X