Buy a new HPLC column, must know the 6 things_

Purchased a brand new HPLC column

Because of the high cost of consumables, most students are afraid of how to unpack a new column.

Today we will introduce the things to note when unpacking a new column!

Note: Since there are many different types of columns, here are some things to note for most of them.

***6 things you must know***

1. Know the maximum pressure, temperature, and pH range of the column

2. How long does it take to equilibrate the column?

3. Solvent preparation

4. line configuration and selection, guard column, and solvent filer

5. Pressure record

6. COA test/storage

1. Know the maximum pressure, temperature, and pH range of the column

Google directly or ask the salesman.

- Maximum pressure resistance: set on the pump to prevent the pressure from causing injury to the column.

- Maximum temperature resistance: special attention should be paid only when there is a temperature box.

- PH value range: To avoid irreversible damage to the tube column caused by too much acid and alkali.

2. How long does it take to equilibrate the column?

According to the volume of the column x 20 times

Ex: I use uHPLCs C18 5um 4.6x250mm as an example

- Inner diameter (diameter): 4.6mm (diameter: 0.46cm) [so the radius is 0.23cm].

- Length: 250mm (25cm)

- Volume formula: radius * radius * 3.14 * length (all use centimeters to calculate)

- V=0.23*0.23*3.14*25=4.15 (ml) (use 4.2ml to do the calculation)

- Please look at the first cell of the table: the volume of the tube column is about 4.2ml, 20 times about 84ml

- If the flow rate is set at 1.0cc/min, it will take about 84 minutes to complete and stabilize

The last line is a new column to run the moving phase of the time required to balance.

3. Solvent preparation

- Use HPLC-grade solvent
- De-aerating with an ultrasonic shaker and filtration with a small flying disk
- Purchase a solvent filter to remove impurities from the solvent

4. configuration and selection of pipeline, guard column and inlet solvent filter

Material: PEEK

Medium pressure resistance; used for pressure zone piping connection; easy to disassemble

I recommend buying the protection column together with the filter head.

From left to right: Inlet solvent filter -> Guard column -> HPLC column

5. Pressure record

- Flow rate should be 0.1cc/min up/down: avoid direct high pressure to the column.

- Table design: My proposed way as follows will make it easy for sales staff and engineers to determine which side is wrong

6. COA test / save

- If you don’t know what COA is, you can refer to this article first

How to determine whether a long-lived tube column is good or bad COA?

Some companies will be very careful to determine whether the column is the same as the original factory. They will play the sample on the COA first to confirm whether there is a difference.

For students who do not have to test, put away the COA paper until the column begins with questions that will come in handy! (If you don’t know, please read that article)

These are the six points that you must know about the new column!

Sometimes there are special notes for special columns; you can also double-check with the manufacturer!