Logo COMPANY NAME
STANDARD OPERATING PROCEDURE | |
Doc No. : ABC/ QC / SOP / 00X | Department: Quality Control |
Revision No: X | Prepared on: xx/yy/zzzz |
Distributed To: QC Department | Effective Date : |
TITLE: OPERATION OF WATERS ALLIANCE 2695 HPLC SYSTEM UV- VISIBLE/ PDA DETECTOR | |
I. PURPOSE:
- To provide a procedure for Operating the HPLC, Make: Waters Alliance equipped with 600 Controller, 2695 Dual absorbance detectors, 2996 Photodiode array detector, and 717 Plus autosampler.
II. SCOPE:
- This procedure applies to the HPLC, Make: Waters, equipped with a 600 Controller, UV-visible detector Waters 2487/2996 Photodiode array detector, and 717 Plus autosampler used in the Quality Control department of ABC Limited. Unit, Location.
III.RESPONSIBILITY:
- It is the responsibility of all the personnel involved in the operation of HPLC, Make: Waters equipped with 695 HPLC, UV-visible detector, 2996 Photodiode array detector, and 717 Plus autosampler.
IV. ABBREVIATIONS:
HPLC : High-Performance Liquid Chromatography
PDA : Photo Diode Array
SOP : Standard Operating Procedure
QA : Quality Assurance
QC : Quality Control.
cGMP : Current Good Manufacturing Practice
Annexures: List of Waters HPLCs.
V. OPERATION:
1.0 WATERS HPLC SYSTEM CONSISTS OF:
1.1 Pump
1.2 In - Line degasser
1.3 600 controller
1.4 UV- Visible Detector Waters 2695
1.5 Photo Diode Array detector Waters 2996
1.6 Autosampler Waters 717 plus
1.7 Check for the calibration status of the instrument. If it is calibrated use it, otherwise calibrate it accordingly and put it into regular use as follows:
1.7.1 Switch on Pump, In-line degasser, 600 Controller, Autosampler and either Dual absorbance detector or PhotodiodeArray Detector depending on the analysis to be performed.
1.7.2 Open the purge valve of the pump and press the home button, this will result in the cursor going to the Flow rate position.
1.7.3 Purge the pump with a flow rate of 5 mL/minute and ensure that no air bubble is stuck up in the line. If exists, repeat the same process until no air bubble persists in the line. After purging, stop the flow and close the purge valve.
1.7.4 Double-click the Empower Login icon on the desktop to login to the waters HPLC system.
1.7.5 Enter the User Name as system and password as manager and click the OK button.
1.7.6 A dialogue box appears which contains
1.7.6.1 Run samples
1.7.6.2 Browse Project
1.7.6.3 Configure system
1.7.6.4 Process data
1.7.6.5 Review data
1.7.6.6 Print data
1.7.7 Click Browse Project. If the desired project already exists, select it and click the OK button to appear in the selected project wizard.
1.7.8 Select the desired chromatographic system by clicking “Tools” and from that Run samples.
1.7.9 In the run samples window, select the desired Instrumental method and click the Setup button to download the method parameters.
1.7.10 To create a new method, click the Edit button load the required parameters as per the standard test procedure and save the method.
2.0 TO MAKE SINGLE INJECTION MODE:
2.1 Go to the View menu, select Single inject, and enter the sample details, such as sample name, function, method set, vial, injection volume, and run time.
2.2 To make an injection, go to the inject menu and select make the injection.
3.0 TO MAKE A SEQUENCE RUN:
3.1 Go to the View menu, select sequence table, and enter the details like Vial, Sample Name, label,
Injection volume, no. of injections Method set Name and run time, save the sequence.
3.2 To run the sequence, go to inject and click Run sample set.
3.3 If the desired project does not exist, click Configure System to create a new one.
4.0 CREATING A NEW PROJECT:
4.1 From the dialogue box that appeared after entering your User Name and Password, click Configure System.
4.2 Click the file menu and select ‘New’ from that Project.
4.3 Enter adequate Table space i.e. 200 MB and then click next.
4.4 From the enabled options select Photo Diode Array, System Suitability and click Next until
Project Name has appeared and save the Project with the desired Project Name.
5.0 CREATING A NEW PROCESSING METHOD:
5.1 Go to the file menu click new method and select “Processing method”
5.2 Select either LC or PDA depending upon the analysis to be performed as the Processing
Type and click ‘OK’
5.3 Enter the appropriate Peak width, Threshold and minimum area in the integration table for optimum analysis of concerned peaks.
5.4 In the components menu enter Peak names and Retention times.
5.5 In the Suitability menu click ‘calculate suitability results’ enter Void volume time depending on the elution and click either US pharmacopoeia, European Pharmacopoeia or all depending on the analysis to be performed and save as appropriate processing method.
6.0 INTEGRATION OF CHROMATOGRAMS:
6.1 Once the analysis was completed double-click on the sample name for the chromatogram to be analysed, which was present in the channels window.
6.2 Then it displays the unintegrated chromatogram, click the file menu and open the appropriate ‘Processing method’
6.3 Go to the process menu and click ‘integrate’. This re-inject results in the integration of the chromatogram.
6.4 Then again go to process and click ‘Quantitate’ to name the components in the chromatogram.
6.5 Then go to ‘File’ click ‘Save’ and select ‘All’ from that. This will result in the save the integrated chromatogram.
6.6 Click ‘File’ and click ‘Exit’ again to close the window.
7.0 TO MAKE A PRINTOUT OF THE INTEGRATED CHROMATOGRAM:
7.1 After completion of integration, click the update button and all the saved chromatograms will displayed in the ‘results’ window.
7.2 Right-click the sample name for the chromatogram to be printed, and select ‘Preview / Publisher’
7.3 On the ‘Open report method’ menu select the appropriate report template
7.4 Click ‘OK’ to print out the concerned chromatogram.
8.0 REFERENCE:
- Operation of Instrument Manual- Waters 2487
8.0 REVISION HISTORY
END OF THE DOCUMENT
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