HPLC Tutorial Part 1_Solvent and Sample Preparations

Mount the nylon filter membrane at the center of the filtration chamber, then carefully attach the upper chamber and secure both components together with the clamp.

Connect the tooth deform to the filtration apparatus and ensure it is tightened properly, as the vacuum pressure is critical for solution filtration.

Pour the mobile phase solution into the compression chamber, switch on the vacuum pump, and allow the solution to pass through the filter until fully filtered.

Remove the filtration assembly and gently place the filtered mobile phase into a clean HPLC bottle. Rinse the bottle as a precaution with a small portion of filtered mobile phase and discard.

Pour the filtered solvent into the prepared HPLC bottle and sonicate the mobile phase to remove any trapped air bubbles in the solvent before analysis.

Label the HPLC vial and inject the prepared sample solution into it, ensuring the sample volume reaches between 0.5 and 1.5 milliliters for proper needle access.

Withdraw the sample into a syringe, attach a membrane filter by twisting clockwise, discard the first 0.5 milliliters of filtrate, then dispense the filtered sample directly into the vial.

Verify that the sample volume is adequate and around 0.5 milliliters, then tighten the vial cap securely to prepare it for HPLC analysis.