Cara Mengoperasikan HPLC atau KCKT

Summary

This video provides a step-by-step guide on how to operate a High-Performance Liquid Chromatography (HPLC) system in a pharmaceutical chemistry lab. It covers the essential preparation, including wearing proper PPE and setting up the equipment, followed by instructions on powering up the system and configuring the software. The video also explains how to prepare the sample, inject it into the system, and monitor the detector reading. Finally, it demonstrates how to properly shut down the system after the analysis is complete. The tutorial is clear, concise, and easy to follow for those new to HPLC operations.

Takeaways

• 😀 Ensure proper preparation before operating the HPLC system, including wearing PPE such as lab coats and masks.
• 😀 Connect the electrical contacts from the stabilizer to the HPLC tool, ensuring the system has power before turning it on.
• 😀 Turn on the HPLC system by powering the pump and detector, followed by the computer used for controlling the HPLC.
• 😀 Open the HPLC application on the computer (LabSolution) and ensure the LCD display is ready for use.
• 😀 Purge the system to remove any air bubbles from the hoses before proceeding with the operation.
• 😀 Make sure the solvent composition for the mobile phase is correctly set and downloaded into the system.
• 😀 Adjust the pump settings and select the desired concentration and composition for the mobile phase.
• 😀 Inject the prepared sample into the HPLC system, adjusting the injection volume and naming the file appropriately in the software.
• 😀 Inject 20 microliters of the prepared sample into the syringe and use the lever to inject the solution into the system.
• 😀 Monitor the chromatogram readings on the detector as the analysis runs, adjusting the flow rate if necessary.
• 😀 Once the analysis is complete, stop the system and turn off the HPLC tool and computer software.

Q & A

What are the initial steps to take before operating the HPLC system?

Before operating the HPLC system, ensure that you are wearing appropriate personal protective equipment (PPE) like gloves, a lab coat, and a mask for safety. Also, check that all necessary components and equipment are prepared and available.

How do you set up the electrical system for the HPLC machine?

First, connect the electrical contacts from the Ki stabilizer to the electricity source. Once this is done, turn on the HPLC machine by powering the pump and detector, followed by the CPU of the computer that will control the system.

What is the first action to take after turning on the HPLC system?

After turning on the HPLC system, launch the LED solution application on the computer to read the data from the HPLC detector. Then, ensure that the LCD screen is ready to use.

Why is purging necessary before running the HPLC system?

Purging is necessary to remove air bubbles from the system or hoses. If air bubbles are not removed, they can interfere with the flow of the solvent, leading to inaccurate results.

How do you ensure the solvent composition is correct before starting the analysis?

Check that the composition of the solvent is correct and that it is ready for use in the HPLC system before turning on the pump.

What do you need to do to configure the HPLC system for sample analysis?

In the software, set the desired concentration of the mobile phase, configure the run as a single run, name the file, and adjust the injection volume and sample type (standard or sample).

How much sample should be injected into the HPLC system?

You should inject 20 microliters of the prepared sample, as specified in the HPLC injection volume settings.

What happens after the sample is injected into the HPLC system?

Once the sample is injected, the system will begin running, and the detector will read the chromatogram. The flow rate of the mobile phase can be gradually adjusted to analyze the sample over time.

How do you stop the HPLC analysis after the sample run is complete?

After the analysis is finished, click on the stop button in the software to end the run, and then turn off the HPLC system and the computer.

Why is it important to adjust the flow rate during the HPLC run?

Adjusting the flow rate allows you to optimize the time analysis and ensure that the chromatogram is accurately recorded, improving the quality of the data collected.