Abstract: High performance liquid chromatography is currently the most widely used chromatographic analysis method. The high performance liquid chromatography system consists of a mobile phase storage liquid bottle, an infusion pump, a sampler, a chromatographic column, a detector, and a recorder. Its overall composition is similar to the gas phase. Chromatography, but a lot of adjustments were made for the characteristics of its mobile phase being liquid.
1). Filter the mobile phase and select different membranes according to your needs.
2). Ultrasonic degassing the mobile phase after suction filtration for 10-20 minutes.
3). Open the HPLC workstation (including computer software and chromatograph), connect the mobile phase pipeline, and connect the detection system.
4). Enter the main menu of the HPLC control interface, click manual to enter the manual menu.
5). It is useless for a period of time, or if a new mobile phase is replaced, you need to flush the pump and injection valve first. Rinse the pump directly at the water outlet of the pump with a needle. To flush the injection valve, under the manual menu, first click purge, then click start, the speed should not exceed 10ml / min when flushing.
6). Adjust the flow rate and use a new mobile phase for the first time. You can try the pressure first. The greater the flow rate, the greater the pressure, generally not more than 2000. Click injure, select the appropriate flow rate, click on, walk the baseline, and observe the situation of the baseline.
7). Design the aliasing method. Click file, select se-lectusersandmethods, you can choose a variety of existing aliasing methods. To create a new method, click newmethod. Select the required accessories, including injection valve, pump, detector, etc., depending on the needs. After selecting, click protocol. A complete sampling method needs to include: a. Steady flow before injection, generally 2-5 minutes; b. Baseline zero; c. Loading-inject conversion of the injection valve; d. Sampling time, depending on the sample different.
8). Inject and operate after injecting. Select the aliasing method and click start. For injection, all samples need to be filtered. After the method is finished, click postrun to record data and mark. After all the samples have been walked, use the above method to walk the baseline for a while to wash off the residue.
9). When shutting down, turn off the computer first, then turn off the liquid chromatography.
10). Fill in the registration book and sign it by the person in charge.
Precautions:
1). Chromatographic purity is required for mobile phase, and 20M deionized water is used for water. The mobile phase after degassing should be vibrated carefully so as not to cause bubbles.
2). The column is very fragile. For the first time, do not let liquid pass through the column.
3). All liquid passing through the column should be strictly filtered.
4). The pressure cannot be too great, it is best not to exceed 2000psi.
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