1. Purpose:
1.1 To ensure the accuracy and reliability of High-Performance Liquid Chromatography (HPLC) results.
1.2. To maintain compliance with regulatory requirements.
2. Scope:
2.1. Applicable to all HPLC instruments in the laboratory.
2.2. Covers calibration procedures for low-pressure quaternary gradient liquid chromatographs.
3. Responsibilities:
3.1. Analyst:
Responsibilities of the Analyst:
3.1. Preparation:
3.1.1. Review the calibration schedule to ensure awareness of upcoming activities.
3.1.2. Gather all necessary materials and standards required for the calibration.
3.1.3. Verify the availability and proper functioning of the required equipment.
3.2. Initial System Check:
3.2.1. Power on the HPLC instrument and allow it to stabilize for at least 30 minutes.
3.2.2. Verify the proper installation of the column and the condition of mobile phase solutions.
3.2.3. Check for any visible leaks or abnormalities in the system.
3.3. Instrument Verification:
3.3.1. Perform an initial system suitability test using a standard sample.
3.3.2. Verify and document the system parameters, including flow rate, pressure, and temperature.
3.3.3. Communicate any irregularities to the supervisor or maintenance personnel.
3.4. Detector Calibration:
3.4.1. Inject a standard solution and optimize detector parameters.
3.4.2. Confirm the accuracy of the wavelength using a certified standard.
3.4.3. Record all detector settings and any adjustments made during calibration.
3.5. Flow Rate Calibration:
3.5.1. Utilize a calibrated flowmeter to measure the actual flow rate.
3.5.2. Adjust the flow rate settings on the instrument based on the measurements.
3.5.3. Document all adjustments made during flow rate calibration.
3.6. Pressure Calibration:
3.6.1. Connect a calibrated pressure gauge to the system.
3.6.2. Compare the gauge reading with the instrument display.
3.6.3. Collaborate with maintenance if pressure adjustments are required and document changes.
3.7. System Suitability Testing:
3.7.1. Run a system suitability test using a standard mixture.
3.7.2. Monitor peak resolution and retention times, ensuring they meet acceptance criteria.
3.7.3. Document all results and report any deviations to the supervisor.
3.8. Documentation:
3.8.1. Record all calibration data, observations, and adjustments in the calibration logbook.
3.8.2. Ensure that entries are clear, complete, and compliant with established procedures.
3.2. Laboratory Manager:
3.1. Planning and Coordination:
3.1.1. Develop and maintain a comprehensive calibration schedule for all HPLC instruments.
3.1.2. Coordinate with analysts to ensure adequate resources, including standards and equipment, are available for calibration.
3.1.3. Review and approve the calibration plan before implementation.
3.2. Resource Management:
3.2.1. Ensure that all necessary standards, reagents, and equipment are in stock and meet quality standards.
3.2.2. Oversee the proper storage and handling of calibration materials to prevent contamination or degradation.
3.2.3. Allocate resources efficiently to minimize downtime during calibration activities.
3.3. Training and Qualification:
3.3.1. Ensure that analysts involved in the calibration process are adequately trained.
3.3.2. Verify that analysts have the necessary qualifications and certifications to perform calibration tasks. 3.3.3. Facilitate ongoing training and professional development for the calibration team.
3.4. Oversight of Calibration Activities:
3.4.1. Review and approve the initial system checks performed by analysts.
3.4.2. Verify that instrument verification, detector calibration, flow rate calibration, and pressure calibration are conducted according to SOPs.
3.4.3. Monitor and address any deviations or issues during the calibration process.
3.5. Documentation Review:
3.5.1. Regularly review calibration logbooks for completeness, accuracy, and adherence to established procedures.
3.5.2. Verify that all necessary signatures and approvals are documented before closing out calibration records.
3.5.3. Investigate and address any discrepancies or non-compliance issues.
3.6. Continuous Improvement:
3.6.1. Periodically assess the effectiveness of the calibration process.
3.6.2. Identify opportunities for improvement and implement corrective actions.
3.6.3. Collaborate with the Quality Assurance team to enhance calibration procedures and ensure compliance with industry standards.
3.7. Communication:
3.7.1. Communicate calibration results and any significant findings to relevant stakeholders.
3.7.2. Ensure that necessary corrective actions are communicated and implemented in a timely manner. 3.7.3. Facilitate a culture of open communication within the calibration team.
4. Materials and Equipment:
4.1. HPLC instrument.
4.2. Certified calibration standards.
4.3. Mobile phase solutions.
4.4. Analytical balance.
4.5. Certified glassware.
4.6. Calibration logbook.
5. Calibration Frequency:
5.1 Factors Influencing Calibration Frequency:
5.1 Manufacturer Recommendations:
5.1.1. Refer to the manufacturer’s guidelines for the recommended calibration frequency.
5.1.2. Take into consideration any specific requirements or recommendations provided by the HPLC instrument manufacturer.
5.2. Regulatory Requirements:
5.2.1. Ensure compliance with relevant regulatory standards and guidelines.
5.2.2. Determine if regulatory bodies prescribe specific calibration intervals for HPLC instruments used in the laboratory.
5..3. Instrument Usage:
5.3.1. Consider the frequency and intensity of use for each HPLC instrument.
5.3.2. Instruments used more frequently or for critical analyses may require more frequent calibration.
5.4. Historical Performance:
5.4.1. Review historical calibration records to identify any patterns or trends.
5.4.2. If there are indications of drift or deviations, adjust the calibration frequency accordingly.
5.5. Maintenance and Repairs:
5.5.1. Take into account any recent maintenance or repairs performed on the HPLC instrument.
5.5.2. Instruments that underwent significant repairs may require more frequent calibration initially.
5.2. Recommended Calibration Frequency:
5.2.1. Monthly Calibration:
5.2.1.1. Recommended for instruments with high usage or critical applications.
5.2.1.2. Ensures continuous accuracy and reliability.
5.2.2. Quarterly Calibration:
5.2.2.1. Appropriate for instruments with moderate usage and stable historical performance.
5.2.2.2. Balances the need for accuracy with operational efficiency.
5.2.3. Semi-Annual Calibration:
5.2.3.1. Suitable for instruments with low usage or consistently stable performance.
5.2.3.2. Ensures compliance while minimizing disruption to routine operations.
5.3. Procedure for Adjusting Calibration Frequency:
5.3.1. Periodic Review:
5.3.1.1. Conduct a periodic review of calibration records and instrument performance.
5.3.1.2. Assess whether any changes in usage or maintenance necessitate an adjustment in calibration frequency.
5.3.2. Risk Assessment:
5.3.2.1. Perform a risk assessment to identify potential consequences of calibration drift.
5.3.2.2. Adjust calibration frequency based on the assessed level of risk.
5.3.3. Documentation:
5.3.3.1. Document any changes in calibration frequency in the laboratory’s calibration schedule.
5.3.3.2. Communicate adjustments to relevant personnel and update the SOP accordingly.
6. Calibration Procedure:
6.1. Preliminary Steps:
6.1.1. Ensure the HPLC system is powered on and stabilized for at least 30 minutes.
6.1.2. Verify that the mobile phase reservoirs are filled with the correct solutions. 6.1.3. Check that the column is properly installed and conditioned.
6.2. Instrument Verification:
6.2.1. Perform an initial system suitability test using a standard sample.
6.2.2. Verify the system parameters, including flow rate, pressure, and temperature.
6.2.3. Record the initial conditions in the calibration logbook.
6.3. Detector Calibration:
6.3.1. Inject a standard solution and optimize detector parameters.
6.3.2. Confirm the wavelength accuracy using a certified standard.
6.3.3. Record the detector settings in the logbook.
6.4. Flow Rate Calibration:
6.4.1. Use a calibrated flowmeter to measure the actual flow rate.
6.4.2. Adjust the flow rate settings on the instrument accordingly.
6.4.3. Document the flow rate adjustments in the logbook.
6.5. Pressure Calibration:
6.5.1. Connect a calibrated pressure gauge to the system.
6.5.2. Compare the gauge reading with the instrument display.
6.5.3. Adjust the pressure settings if necessary and record changes.
6.6. System Suitability Testing:
6.6.1. Run a system suitability test using a standard mixture.
6.6.2. Ensure that peak resolution and retention times meet acceptance criteria.
6.6.3. Document the test results in the logbook.
7. Documentation:
7.1. Documentation Preparation:
7.1.1. Calibration Logbook:
7.1.1.1. Maintain a dedicated calibration logbook for each HPLC instrument.
7.1.1.2. Record instrument identification, date, and details of the calibration activities.
7.1.2. Standard Operating Procedures (SOPs):
7.1.2.1. Ensure all personnel are familiar with the relevant SOPs for calibration.
7.1.2.2. Document any deviations from the SOP and the justification for such deviations.
7.2. Pre-Calibration Documentation:
7.2.1. Calibration Plan:
7.2.1.1. Develop a calibration plan specifying the instruments, standards, and equipment to be used.
7.2.1.2. Include details of the calibration frequency and any specific requirements.
7.2.2. Material Verification:
7.2.2.1. Document the verification of calibration standards and reagents.
7.2.2.2. Confirm the expiration dates and traceability of all materials used.
7.3. Calibration Activities Documentation:
7.3.1. Initial System Checks:
7.3.1.1. Record the results of the initial system checks performed by the analyst.
7.3.1.2. Document any observations, abnormalities, or adjustments made.
7.3.2. Instrument Verification:
7.3.2.1. Document the parameters verified during the system suitability test.
7.3.2.2. Record any deviations and corrective actions taken.
7.3.3. Detector Calibration:
7.3.3.1. Record detector calibration parameters and adjustments made.
7.3.3.2. Include details of the standard solution used for calibration.
7.3.4. Flow Rate Calibration:
7.3.4.1. Document the flow rate measurements using a calibrated flowmeter.
7.3.4.2. Record any adjustments made to achieve the desired flow rate.
7.4.5. Pressure Calibration:
7.4.5.1. Document the results of pressure calibration using a calibrated gauge.
7.4.5.2. Record any adjustments made to meet specified pressure requirements.
7.5.6. System Suitability Testing:
7.5.6.1. Record the results of the system suitability test.
7.5.6.2. Include details of peak resolution, retention times, and other relevant parameters.
7.4. Post-Calibration Documentation:
7.4.1. Review and Approval:
7.4.1.1. Ensure that the calibration logbook is reviewed by the Laboratory Manager.
7.4.1.2. Obtain necessary approvals and signatures to confirm completion.
7.4.2. Deviation Reports:
7.4.2.1. If deviations occurred, prepare a deviation report.
7.4.2.2. Include details of the deviation, its impact, and corrective actions taken.
7.5. Record Retention:
7.5.1. Maintain calibration records for a specified retention period as per regulatory requirements.
7.5.2. Store records in a secure and accessible location.
7.6. Training:
7.6.1. Ensure all personnel involved in calibration activities are trained on proper documentation procedures.
7.6.2. Periodically review and update training programs.
7.7. Review:
7.7.1. Periodically review this SOP to ensure alignment with industry standards and regulatory requirements.
7.7.2. Update the documentation process as needed based on new information or changes in laboratory procedures.
7.8. Approval:
7.8.1. This SOP is approved by the Laboratory Manager.
7.8.2. Review and approval shall be documented in the SOP.
8. Training:
8.1. Ensure that all personnel involved in HPLC calibration are adequately trained.
8.2. Periodically review and update training programs.
9. References:
9.1. Manufacturer’s manual for the specific HPLC instrument.
9.2. Relevant industry standards and regulations.
10. Review:
10.1. Periodically review and update this SOP as needed.
10.2. Conduct internal audits to ensure compliance.
11. Approval:
11.1. This SOP is approved by the Laboratory Manager.
11.2. Review and approval shall be documented in the SOP.
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