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SOP for Operation of Shimadzu HPLC LC-2010 CHT

OBJECTIVE:
To provide guidelines for the operation of the HPLC SHIMADZU LC-2010CHT system.

PURPOSE:
To lay down a procedure for the operation of the HPLC SHIMADZU LC-2010CHT system.
RESPONSIBILITY: 
HPLC operating chemist / Departmental Head.

PROCEDURE:
  • Switch on the instrument and after initialization enter the password menu will be displayed.
  • Purging the system before start of analysis.
  • Click on purging on the display.
  • Press A, B, C, D & INJ on display.
  • Keep the column into the system.
  • Then go to menu by pressing the cursor ▲ or ▼.
  • Set the system for column washing to the initial composition of A = Mobile phase 100%, B =Methanol 100 %, C = Acetonitrile 100% & D = Water 100% and put column oven @25 to 60ºC and put flow at 1.0ml/min and press the Pump button.
  • Set the flow at 1.0 ml/min leave for 1min and check for leaks. Leave the system to equilibrate for 2 min.
  • Instrument ready for analysis.

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Connecting to the LC Solution software.
  • Click → LC Solution icon.
  • Click →Administration
  • Click →User Administration.
  • User ID – Admin → ok (by default).
  • Method Creation.
  1. Go to → Assistant Bar.
  2. Click → Top.
  3. Click → Data Acquisition.
  4. Click → Instrument Parameters.
  5. Click → Advanced.
  • Data Acquisition→ LC stop time 25 min and then click apply to all Acquisition time.
  • LC time prog
  • Gradient flow → Total flow at 1.0 ml/min.(Adjust the parameters as per the method requirement)
  • Pump → Mode → Isocratic flow and Gradient flow.
  1. Isocratic flow → Flow at 1.0 ml/min, Port – A and pressure limits Maximum - 380 kgf/cm2, Minimum - 0 kgf/cm2.
  2. Gradient flow → Total flow at 1.0 ml/min. (Adjust the parameters as per the method requirement)
  • Detector-A
  1. Lamp-D2
  2. Polarity-+
  3. Response-1.0 sec
  4. Cell temperature- Low
  5. Wavelength-254 nm. (Adjust the parameters as per the method requirement)
  • Column oven → No need to change.
  • Controller → No need to change.
  • Autosampler
  1. Sample Rack-1.5 ml 70 vials- Click-Detect Rack.
  2. Rinsing volume -200 ul.
  3. Needle stroke-52 mm.
  4. Sampling speed-15 ul/sec.
  5. Rinse mode – Before and after Aspiration.
  6. Rinse Dip time – 0.
  • Auto purge → No need to change.
  • Go to file → save method file as Batch No. and then click Download.
  • Click → Batch processing, click, then go to file new batch file → Vial Right click, Add Row → select → click → number of samples-10 ok.


Report Format Creation
  • Click Postrun → click Report format → click file → new report format file.
  • Then go Item → General Text one window will come → Text type the company name → go general Font setting → Times new → Bold → ok → apply → ok.
  • Click Item → common → sample information.
  • Click Item → LC/PDA common → Chromatogram.
  • Click Item → LC/PDA common → Peak table → go to file save report format.

Integrate the Data File
  • Click Postrun → click Top → LC Data analysis→ select browse folder → elegant select.
  • Select the data file → double click → click wizard → Program → Reject peaks → after integration→ click simulate then ok.

Calculate the % RSD for Area and RT
  • Top → click report format → click file → new report format file → Item → common → sample information.
  • Item → LC/PDA common → Summary compound.
  • Click right properties then go file → add → click table.
  • Print the required peak and close.
  • Printing of the chromatogram.
  • Click on chromatogram → preview and publish → window opens → select individual report → ok → print.
  • Column washing after sample sequence completed.
  • When the sequence is finished, stop the flow by pressing “Stop pump” button.
  1. Turn off the instrument.
  2. Use Annexure to check the column performance details.

REVIEW HISTORY:
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1 Comments

  1. Mr.Patel
    It is showing excellent
    Keep it up for helping us with
    Other formulation improvement.

    ReplyDelete

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