High-performance liquid chromatography or HPLC is a method of column chromatography in which the mobile phase (a sample mixture or sample dissolved in solvent) is pumped through a stationary phase (typically a column packed with chromatographic material) at high pressure.
HPLC is used to identify, quantify, or purify the individual compounds within a mixture. The sample is moved through the system using a carrier gas stream. The retention time (the amount of time that the analyte takes to move through the system) of each sample varies depending on the compound being analyzed, the solvent used, and the stationary phase.
HPLC (High-Performance Liquid Chromatography) preventive maintenance is critical to preserving the accuracy, precision, and dependability of this analytical technology. It consists of a set of routine inspections, calibrations, and cleaning operations designed to detect and avoid possible issues before they arise.
Proper HPLC preventative maintenance can assist in eliminating downtime, reducing the need for repairs, extending the instrument's lifespan, and guaranteeing that the HPLC findings are correct and consistent over time. Inspecting and cleaning the sample injection system, examining the pump for leaks or anomalies, validating the flow rate accuracy, and evaluating the detector's sensitivity and wavelength accuracy are some of the usual duties involved in HPLC preventative maintenance.
Overall, preventative maintenance is an important component of any laboratory's quality control program, and frequent attention to HPLC maintenance may assist in guaranteeing that the instrument continues to perform ideally and reliably for years to come.
ALSO READ: Troubleshooting Guide – HPLC
OBJECTIVE
This SOP describes the procedure for Preventive Maintenance of High-Performance Liquid Chromatography.
RESPONSIBILITY
Executive – Quality Control– To perform the preventive maintenance activity.
Head – Quality Control – To ensure compliance with the established schedule.
PROCEDURE
Precautions:
- Ensure that the power supply to the instrument is switched “off” before starting the preventive maintenance.
- Clean the instrument with a clean dry cloth every day.
- Performance checks shall be done for the following parameters as per Calibration sop After doing preventive maintenance
- Enter the values in the standard maintenance card as per the SOP Preventive Maintenance procedure for Laboratory Instruments.
- The following components of HPLC shall be subjected to preventive maintenance on the scheduled date.
General cleaning:
- Before starting the cleaning you must ensure the power supply to the HPLC system should be switched off.
- Clean the instrument with a clean dry cloth every day.
Performance Verification:
A. Solvent Delivery Module
- Check flow accuracy as per SOP
- Acceptance criteria: ± 2 % of set flow rate.
B. Detector
- Check the Detector Wavelength accuracy as per SOP.
- Check the Detector Wavelength accuracy as per SOP.
C. Injector
- Check the Injector Linearity Precision as per the SOP.
Acceptance Limit is:
- The % RSD for all peak areas should be NMT 1.0.
- The correlation coefficient should be NLT 0.999.
Procedure for Preventive Maintenance:
Pump Solvent Separation Module
- Switch off the main power switch.
- Clean the front panel and cover
- Sonicate solvent inlet filters, drain valve filter inlet, and outlet check valves in methanol for 5 min.
- open the pump head check the plunger and plunger seal for scratches on the plunger and replace if necessary
- Check seal wash assembly seals, and replace if necessary.
- Check and sonicate inlet valve cartridges
- Check pump functioning for pressure fluctuations.
Detector
- Remove dirt from the panel and cover.
- Reassemble cell and Sonicate flow cells with methanol or IPA for 10 minutes.
- Fix the flow cell back after sonication.
- Reassemble cells, and check for leaks.
- Perform automatic wavelength calibration.
- Check sample energy and reference energy, Replace the lamp if necessary
- Check for leak sensor functioning.
- Stick the preventive maintenance label before doing preventive maintenance
- Frequency:- After each maintenance
Auto-Injector
- Check the injector syringe, and replace it if necessary
- Clean all tubes in the injector
- Perform the purge and compression ensures all fitting are tight.
- Adjust the seal pack if necessary to optimize the needle port.
- Check the needle and the seal pack seal replace if necessary.
Column Oven
- Remove dirt from the panel and cover
- Check the function of the circuit cooling fan, and replace it if it is not working.
- Check column oven functioning
- Check leak sensor functioning
- Check tubing connections for damages, cut tubing, and replace new tubing if Necessary.
Enter the preventive maintenance checks in the standard maintenance card as per Stick the preventive maintenance label before doing preventive maintenance as per SOP.
The frequency of the schedule is every 3 months.
ABBREVIATIONS
SOP – Standard Operating Procedure
HPLC – High-performance liquid chromatography
QC-Quality Control.
ANNEXURES
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