Agilent Chemstation Software Manual

1.0 OBJECTIVE: To lay down the procedure for operation and calibration of High Performance Liquid Chromatograph. 2.0 INSTRUMENT IDENTIFICATION: Name of Instrument: High Performance Liquid Chromatography (Auto Sampler) Manufacturer: AGILENT Technologies 1260 series 3.0 GENERAL CLEANING: 3.1 Ensure that the power supply to the instrument is switched off and main Cord is removed from supply.

3.2 Clean the instrument with a clean dry cloth every day. A wet cloth dipped In dilute soap solution may be used occasionally. 3.3 Precaution must be taken to clean the instrument immediately with dry Cloth after wet cleaning. 4.0 PROCEDURE (OPERATION) 4.1 Ensure that the HPLC system is placed in a clean and dust free area. 4.2 Ensure that instrument have been connected to the main power unit.

4.3 Switch ‘ON’ the Computer. 4.4 Switch ‘ON’ the HPLC instrument. (Switch on the detector, column compartment, Pump and vacuum degasser simultaneously). 4.5 Ensure that the outlet from the detector dips into a waste bottle.

5.0 Start the Open Lab Chemistration by double-clicking the Short cute Open Lab Chemistration located on your desktop. 5.1 Click on Open Lab Chemistration online. 5.2 Then click on ‘on’ option.

Agilent and the user have a separate. In this manual we describe the efficient use of the data acquisition, analysis. About the ChemStation Software 12.

5.3 Initialization process of HPLC system will be start which shows by yellow colour. 5.4 Green colour and ‘Ready’ indication will be appearing on screen after the Initialization process. 5.5 Click on set up- pump, feed the flow rate NMT 4%,% of solvent and then start the purging by rotating the purge wall in anticlockwise direction approx 10 minute. 5.6 Then ‘OFF’ the Purging by rotating the purge valve in clockwise direction.

Make sure the sample to be run is properly loaded in the auto sampler your instrument. 5.7 Start an instrument application an Instrument Wizard will appear. This wizard is designed direct to the basic functions of the instrument.

5.8 Create or modify a method This button starts the Method Wizard that will enable you to step through creating or modifying a method. 5.9 When you start the Method Wizard, click to create a new method the wizard will Guide you through the method sections. For each section, enter the run Parameters Required for the sample you will be injecting 5.10 Data acquisition and instrument control parameters – (a) Instrument Setup. Select each tab and enter the correct parameters for running this sample (b) Integration events. Additional integration events are not necessary for a test Sample.

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Calibration of Peaks/Groups. Calibration parameters are not necessary for a test sample. From the File menu, select Report Template and then click Open.

From the list of templates, select Area%.rep to generate an Area%report when run the sample 5.11 Create a sequence This button starts the Sequence Wizard that steps you through creation of an acquisition reprocessing sequence. Or for creating sequence enter file menu which display Sequence, from Sequence enter new method. 5.12 Run one sample this button opens a dialog where you can use a stored method to run a single sample.

• In the Single Run Acquisition dialog box, enter a number to be used as a Sample ID. • Click the method open button and then select the method file that was provided or the method you saved in the previous step. • Enter a Data file name you wish to use, or click the adjacent button to select a parameter that will be used to create the data file name for you. • Enter the Vial number and Injection Volume to be used.

5.13 Run sequence of samples this button opens the Run Sequence dialog where you can start data acquisition using a stored sequence. 5.14 Save the method. When you have completed setting up the method parameters, from the File menu, followed by Method and click Save As. Type a name to be used for your method and then click OK 5.15 Maintain Log book of HPLC 5.16 Report any discrepancy observed during operation and calibration of instrument to Section In charge or his representative for corrective and preventive action. 5.17 Section In charge or his representative will take the necessary action and report the same to Manager – Quality Control.