Core principle: Follow the five step process of "preparation → activation → sample loading → rinsing → elution", without the need for complex debugging. Focus on controlling column activation and flow rate control, which beginners can master in 10 minutes.

Applicable scenarios: Enrichment and purification of target substances in environmental water samples, food samples, biological fluids, and other samples (taking common solid-phase extraction columns as an example).

1、 Preparation before operation (2 minutes, key pollution prevention)

Instrument inspection: confirmSolid phase extraction instrumentThe host is powered on and the vacuum pump (or pressure pump) is connected normally; Check if the extraction column bracket and collection tube bracket are securely installed, and place the corresponding collection tube (centrifuge tube/test tube) on the bracket.

Preparation of reagents and materials: Prepare solid-phase extraction columns (such as C18 columns, cation exchange columns), activation solvents (such as methanol), elution solvents (such as 5% methanol aqueous solution), elution solvents (such as pure acetonitrile), and samples to be processed (filter out particles in advance to avoid column blockage).

Column fixation: Insert the solid-phase extraction column vertically into the instrument bracket, aligning the lower end with the collection tube; If it is a manually adjustable instrument, tighten the column fixing clamp to ensure that the column is stable without shaking.

2、 Five step core operation (6 minutes, precise speed control is key)

Step 1: Column activation (activation of adsorption sites, must do!)

Use a pipette to aspirate 3-5 mL of activated solvent (such as methanol) and slowly inject it into the extraction column;

Turn on the vacuum pump (or start the pressure pump) of the solid-phase extraction instrument, adjust the flow rate to 1-2 mL/min (too fast a flow rate can cause insufficient activation), and turn off the vacuum pump after the solvent is completely dried (to avoid dry burning of the column);

If the extraction column is compatible with aqueous phase, 3-5mL of ultrapure water should be used for rinsing, and the flow rate should be controlled at 1-2mL/min. After drying, it should be used for later use.

Step 2: Sample loading (adsorption of target substance)

Take the preprocessed sample (usually 5-20mL, depending on the column capacity) and slowly inject it into the activated extraction column using a pipette or sampler;

Turn on the vacuum pump and adjust the flow rate to 1-3mL/min (adjust according to the viscosity of the sample, and slow down slightly for turbid samples) to ensure that the sample flows evenly through the column and the target substance is fully adsorbed;

After the sample is loaded, continue pumping for 30 seconds to dry the residual sample in the column.

Step 3: Rinsing (removing impurities and retaining the target substance)

Take 3-5 mL of elution solvent (such as 5% methanol aqueous solution, choose according to the properties of the target substance to avoid elution) and inject it into the extraction column;

Maintain a flow rate of 1-2 mL/min to allow the rinsing solvent to dry. During this process, a new collection tube can be replaced to avoid impurities contaminating the subsequent eluent;

After rinsing, evacuate for 1 minute to thoroughly remove any residual rinsing solution from the column.

Step 4: Elution (collecting target substance, core step)

Align the clean collection tube with the lower end of the extraction column, draw 1-3 mL of elution solvent (such as pure acetonitrile, ensure complete elution of the target substance), and slowly inject it into the column;

Adjust the flow rate to 0.5-1mL/min (if the flow rate is too slow, the efficiency will be low, and if it is too fast, the elution will not be complete), allowing the eluent to flow completely into the collection tube;

The elution can be repeated once (if the recovery rate of the target substance is low), and the two elution solutions can be combined to complete the extraction.

Step 5: Final Cleaning (Extend Instrument Life)

Turn off the vacuum pump and instrument power, remove the used extraction column, and place it in a dedicated waste container;

Rinse the extraction column bracket and pipeline with ultrapure water (if there is residual solvent), and then wipe them clean with a dry cloth;

Organize the reagents and collection tubes, and place the instrument in a dry and ventilated place.