1.0 Purpose:
1.1 This Standard Operating Procedure (SOP) outlines the systematic procedure for the calibration of High-Performance Liquid Chromatography (HPLC) equipment, specifically the Low-Pressure Quaternary Gradient Liquid Chromatograph, to ensure accurate and reliable analytical results.
2.0 Scope:
2.1 This procedure is applicable to all HPLC instruments utilizing a Low-Pressure Quaternary Gradient system within the [Your Company Name] laboratory.
3.0 Responsibilities:
3.1 The Analyst –
4.0 Equipment and Materials:
4.1 HPLC instrument with a Low-Pressure Quaternary Gradient system.
4.2 Certified standard solutions for calibration.
4.3 Graduated cylinders.
4.4 Analytical balance.
4.5 Chromatographic columns.
4.6 Mobile phase solutions.
4.7 Injection syringes.
4.8 HPLC-compatible solvent.
4.9 Calibration report template.
5.0 Procedure:
5.1 Preliminary Checks:
5.1.1 Ensure the HPLC instrument is clean and free from any contamination.
5.1.2 Verify that the instrument is in the standby mode and is properly connected to the power source.
5.1.3 Check the condition of the chromatographic column and replace if necessary.
5.2 Instrument Warm-up:
5.2.1 Turn on the HPLC instrument and allow it to warm up for a minimum of 30 minutes.
5.3 System Suitability Test:
5.3.1 Run a system suitability test using a standard solution to ensure the proper functioning of the HPLC system.
5.4 Calibration of Detectors:
5.4.1 Calibrate the UV/Visible and other detectors according to the manufacturer’s specifications.
5.5 Calibration of Pumps:
5.5.1 Prime the pumps with the HPLC-compatible solvent.
5.5.2 Calibrate the flow rate and pressure using certified standard solutions.
5.6 Calibration of Injection System:
5.6.1 Load the injection syringe with a known concentration of the standard solution.
5.6.2 Inject the standard solution and ensure proper peak shape and retention time.
5.7 Calibration of Gradient System:
5.7.1 Prepare a set of standard solutions covering the desired concentration range.
5.7.2 Run a gradient program using the standard solutions and verify the linearity of the detector response.
5.8 Data Analysis:
5.8.1 Analyze the chromatograms obtained during the calibration for peak resolution, symmetry, and accuracy.
5.8.2 Document the results in the calibration report template.
6.0 Documentation:
This Standard Operating Procedure (SOP) outlines the specific guidelines for the comprehensive documentation of the High-Performance Liquid Chromatography (HPLC) equipment calibration process, emphasizing the Low-Pressure Quaternary Gradient Liquid Chromatograph system.
7.0 Training:
7.1 Ensure that all personnel involved in HPLC calibration are adequately trained and understand this SOP.
8.0 References:
8.1 Manufacturer’s manuals for HPLC instrument and components.
9.0 Revision History:
| Revision No. | Description | Date |
|---|---|---|
| 1.0 | Initial release | [Date] |
10.0 Approval:
10.1 [Name and Signature of Approving Authority]
For more articles, Kindly Click here.
For pharmaceutical jobs, follow us on LinkedIn
For Editable SOPs in Word format contact us on info@pharmaceuticalcarrier.com
For more information kindly follow us on www.pharmaguidelines.co.uk
Download Above SOP