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SOP for Operation of Shimadzu HPLC (Prominence – i LC – 2030)

OBJECTIVE:
To lay down a procedure for the operation of HPLC system using Lab Solution software. 

SCOPE:
This is applicable for the operation of HPLC system using Lab Solution software for Shimadzu Prominence – i LC – 2030 HPLC system. 

RESPONSIBILITY:
It is the responsibility of QC Executive. 

ACCOUNTABILITY:
Manager – Quality Control 
Manager – Quality     
PROCEDURE:
Operation of High Performance Liquid Chromatography
  • Before starting analysis, ensure that the HPLC System is placed on a clean and dust free platform. 
  • Ensure that the instrument has been connected to power supply. Ensure that the all communication cords between the units are connected properly. 
  • Switch on the instrument (Including Injector, Pump, Column Oven and UV lamp) by clicking the “ON” button on the extreme left middle corner of the screen and screen will be change red to green. Initialization will occur for 3 - 5 minutes; self-checkup of the instrument will occur. 
  • Ensure that appropriate solvent mixture (water: Methanol) (70:30, v/v) is pumped for seal wash. 
  • Ensure that the outlet tube from the detector is dipped into a waste bottle. 
  • Connect the column specified in the individual monograph in column oven compartment in the flow direction marked on the column. Initially, wash the column by flushing with water: Methanol (70:30) or appropriate solvent if specified in the monograph which is filtered through 0.45 µ filter for at least 30 minutes. 
  • Prepare the mobile phase as mentioned in the monograph of specific product. Fill it in the reservoir bottle. Suspend the inlet tube into the reservoir bottle and ensure that the filter is below the level of mobile phase. 
  • After the completion of self-diagnosis, the main menu of work station is displayed on HPLC Screen. 
  • Double click the Lab Solution icon on desktop screen to open the Lab Solution software & Logging in. 
  • Click on Instrument à HPLC – 2030 à Click on Instrument ON at down assistant Bar. 
PEOPLE ALSO READ: SOP for System Suitability Test in HPLC analysis and HPLC Documentation

Procedure for Method Creation:
  • Click on Instrument à HPLC – 2030 à Go to File à New Method File à Create Method as per Monograph à Download & Close à File à Save Method File As à Save in Specific Product Folder. 
  • Open the purge valve of the instrument and purge the corresponding solvent line (s) in which mobile phase have been placed by setting the flow rate of the pump as 1.0 ml/min for few minutes until the solvent line is completely filled with new mobile phase. 
  • Ensure that no air bubbles are noticed in the solvent line. 
Procedure for Batch Creation:
  • Click on Main at Left Assistant Bar à Real Time Batch à Go to File à New Batch File à Input Data à File à Save Batch File As à Save in Specific Product Folder. 
Procedure for Single Run:
  • Click on Acquisition at Left Assistant Bar à Start Single Run à Input Require Detail à Press OK à Run the Method & Screen change green to dark blue. 
Procedure for Sequence Run:
  • To Run Batch, Click on Main at Left Assistant Bar à Real Time Batch à Start Real time Batch 
  • The instrument run automatically after some time. 
  • While the sequence is running, chromatogram can be viewed online from the postrun. 


Procedure for Data Analysis:
  • Click on Data explorer sub-window, & double click on specific chromatogram. 
  • Click on “Edit” to edit each parameter value. 
  • Click on “Integration”. Input Width 5 sec & Slope 200 uV/min. 
  • Click on “View” to perform processing on the data, & the processing results are displayed in Chromatogram view and result view – peak table. 
  • Click on “Edit”. Clik Quantitative. For quatitative method select ‘External Standard’. 
  • Select 1 for # of calib. Levels. 
  • Select Conc. for X axis of calib. Curve. 
  • Click on “Compound”. Selecting the retention time cell, & clicking the peak in chromatogram view automatically enters the retention time of that peak to the currently selected [Ret. Time] cell. The retention time can be set by simply clicking the mouse. 
  • Click on ‘save’ from top assistant bar. 
  • Click on ‘Apply to Method’ to save method file. Tick on ‘current setting’ & ‘Data processing parameters’. 
  • Click ‘OK’ to save the new data processing parameters. 
  • Click on Batch sub-window & open perticular batch. In method file column add new data processing parameters method & fill down. Save batch. 
  • Select batch & click on Start. Data processing will start. 
  • Click on the “Report Format” icon. Open report format. The final report is displayed according to the report style selected. 
  • Adjust the scale of the chromatogram if required by selecting chromatogram properties. The window of the chromatogram properties will have an option for time scale and chromatogram scale which are adjusted as required. Click OK to close this window. 
  • Save any changes made in report format. Click on Print icon at the top of the screen to print the chromatogram. To view the next chromatogram, drag and drop other chromatogram and take print out similarly. 
Procedure for System Shutdown:
  • After completion of analysis, wash the column by downloading washing method & keep it back in the relevant place. 
  • Click on ‘Instrument ON’ at down assistant Bar to disconnect instrument form Pump, Lamp & Column oven. 
  • Close the panel by click on file & then exit. 
  • Switch off small round button & switch off the mains of system controller. After completed switch off the PC. 
  • Enter the column & instrument usage details in the logbook. 

ABBREVIATIONS:
HPLC - High Performance Liquid Chromatography 
UV - Ultra Violet 
PC - Program Controller[full_width]


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