HPLC Maintenance and Care

1. Manual sampler:

Usually should pay attention to the use of secondary distilled water and methanol in the loading state and injection analysis state cleaning; If there is leakage phenomenon, the reason is likely to wear or contamination of the rotor gasket, generally apply for repair or replacement parts.

2. Drain valve:

Here can not be completely sealed or leakage of the gasket is generally one of the contamination or wear, can be removed after the removal of its gasket with isopropyl alcohol ultrasonic cleaning or replacement of the gasket.

3, check valve:

If the pump pressure is unstable or the mobile phase cannot be pumped, there may be a problem with the one-way valve. Remove the ultrasonic cleaning with isopropyl alcohol. When cleaning, place it in a small beaker according to the direction in which it is installed. Remember not to install it. Invert super wash!

4, workstations:

Deadlock can restart the computer; when it is not running properly, it can be replaced by computer to test its hardware failure; or re-plug the interface on the machine, re-install the software.

5, filter:

When abnormality occurs in the chromatographic peak, there is a possibility that the component is contaminated. Remove the isopropyl alcohol or replace the filter gasket.

6, suction filter head:

Under special circumstances, the filter head can be removed to determine whether it is clogged. It is also possible to use a syringe to suck the mobile phase to determine if it is blocked by the suction filter. If there is a blockage, isopropyl alcohol can be used for ultrasonic cleaning; if it is unsuccessful, it needs to be replaced.

7. Extraction requirements for mobile phase:

High performance liquid chromatography alcohol, double distilled water, buffer salt must be filtered;

Degassing of the mobile phase is critical (can use suction filtration, ultrasonic degassing and other methods)

8, the flow pool:

When the chromatographic peak is not normal, this part may be contaminated. The detachable gaskets are removed and washed with isopropyl alcohol (can be operated according to the instructions or repaired).

9, pump head:

Leakage or other failure of the pump head is generally required to apply for maintenance (such as leakage, or replace the plunger rod and its seals, etc.)

Unstable pump pressure:

1. The pump head has air bubbles - mobile phase degassing / large flow flush pump / pumping fluid flow with a syringe

2, check valve failure - cleaning

If the above operation still cannot be solved, use isopropyl alcohol to flush the flow (no column flushing, directly into the detector from the manual injector), flow rate is 3-4ml/min, rinse for about 50 minutes, then reinstall the column and replace it. Balanced mobile phase, so it can not solve the problem of pump pressure fluctuations, can apply for replacement parts.

HPLC maintenance:

Leakage treatment:

The C18 column must not contain protein samples, blood samples, or biological samples.

Pay attention to the pH range of the column and do not inject samples of strong acids and alkalis, especially alkaline samples.

If the instrument is not used for a long time, the column should be sealed and sealed with a plug. Note that the column should not be preserved with pure water, but should be preserved with an organic phase (such as methanol) because the pure water is prone to mildew.

Blocking of the flow path: The pressure caused by the blockage is too high. Press the pre-column → filter in the mixer → line filter → check valve and clean it. Cleaning methods: 1 with civilian propanol as a solvent rinse 2 in isopropyl alcohol in ultrasonic cleaning 3 with 10% dilute nitric acid cleaning

Use of buffer salts:

When the mobile phase contains salt, the flow path must be flushed after the experiment is finished;

First flush with water, open the drain valve and use the PURG key to convert the original salt-containing mobile phase, then close the drain valve and open the pump to flush the entire flow path for more than 45 minutes. Replace the mobile phase so that the mixture of water and methanol flushes the entire flow path. 30 minutes (this step can be generally omitted, according to the experiment), the last replacement of pure methanol flushes the entire flow path for 45 minutes or more.

Pump head flushing;

Rinse 3-5 ml with distilled water and pure methanol with needles and syringes in spare parts.

If the mobile phase does not contain salt, the machine can be regularly flushed and maintained, depending on the number of experiments.

Manual sampler flushing;

Also, use a syringe in the spare part and a special flushing needle to flush the 3-4 ml of distilled water with the loading status and injection status of the manual injector, and then rinse with 3-4 ml of pure methanol. Make sure that every time you finish the experiment, all flow paths are full of pure methanol!



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