OPERATION OF HIGH PRESSURE LIQUID CHROMATOGRAPHY

1.0. PURPOSE:
To provide procedure for Operation of High Pressure Liquid Chromatography.
2.0. SCOPE:
Applicable to HPLC (CodeNo. ) :
Make : Shimadzu
Model : LC-2010A
Pump : LC-10Atvp Detector : SPD-10Avp
System Controller : SCL-10Avp
Auto Injector : SIL-10Advp
3.0. RESPONSIBILITY:
3.1. QC- officer
3.2. QC- Executive
4.0. ACCOUNTABILITY:
Manager-Quality Control
5.0. DEFINITION:
Nil
6.0. PROCEDURE:
6.1.1. Check and ensure that the instrument is clean and calibration is valid.
6.1.2. Switch on SCL10Avp system control,SPD 10Avp UV-Visible detector, followed bySQL/10ADvp autoinjector & followed by LC10ATvp pump.
6.1.3. Then start the computer (Softwere),wait for a while;Till enter Password, appear on the monitor.
6.1.4. Enter the password -On screen WELCOMES TO WINDOWS appear.
6.1.5. Enter Esc wait for a while.
6.1.6. Icons appear. Then double click on class-vp system & wait for a while.
6.1.7. Class -vp system will open.
6.1.8. Choose Auto Sampler with double click.
6.1.9. On monitor login appear,enter the users name & Password . Enter User name as System , Password as 2001
6.1.10. Then Icons appear on the screen click on File,choose for METHOD, open a New file or stored file & same by going back to method.
6.1.11. Once again go back to file, Choose the SEQUNCE & save,enter file name data name & save.
6.1.12. Next go to METHOD icon on the screen click over method.
6.1.13. Choose INSTRUMENT SETUP then PUMPS/ SPD 10 AVP (Detector-A) will appear.
6.1.14. Set the Pump for flow rate & max pressure.
6.1.15. In SPD10VP (Detector-A),choose for wavelength & click APPLY followed by DOWNLOAD & save the method.
6.1.16. Once again come back to main screen,then choose icon on the main screen.
CONTROL,run down click over DIRECT CONTROL the series of icons appear on the screen.
6.1.17 Click icon on instrument ON/OFF Now the system is ready for operation Once analysis is over close the system as follows.
6.1.18. Click over file run down to EXIT.
6.1.19. Click once again on screen SHUTDOWN the computer,it's now safe to turn off your computer message appear.
6.1.20. Similarly switch off SCL10AVP system controller, followed by SPD10AVP, UV-VISIBLE DETECTOR followed by Auto-Injector, & then LC10ATVP pump.
Acceptance criteria : maximum pressure variation must not exceed 5.0 Kgf/cm2.
6.1.21. SELECTION OF COLOUMN: Check for the coloumn specified in the individual Monograph, and wash properly with solvents specified to get the proper resolution.
6.1.22 SYSTEM SUITABILITY: Carry out the system suitability test mentioned in individual Monograph.
6.1.23 ‘Frequency of washing’ : Column washing should be carried out after each analysis .
6.1.24 Extra washing should be carried ,whenever abnormal pressure variation or distortion of peaks occurs during analysis.
7.0. REFERENCES:
Instrument Manual
8.0. ENCLOSURES:
HPLC usage log
No.SOP:QCOP:00x6:xx
9.0. ABBREVIATIONS:
Nil