Cleaning method of liquid chromatography column

High performance liquid chromatography (HPLC) is a separation and analysis technology developed in the late 1960s. It is an important means of modern separation and determination. Since its advent, it has been widely used in biochemistry, medicine and clinical analysis due to its high separation efficiency, fast analysis speed, good detection sensitivity and analysis of thermally unstable physiologically active substances with high boiling point but can not be gasified. High performance liquid chromatography (HPLC) column is a consumable. With the increase of using time or injection times, the peak height, peak width and column efficiency will decrease. Regular thorough cleaning and regeneration are required. Different cleaning methods of chromatographic column are different, such as:
1. Reversed phase chromatographic column
Methanol: water = 90:10, pure methanol (HPLC grade), isopropanol (HPLC grade), dichloromethane (HPLC grade) and other solvents were used as mobile phases. Each mobile phase flowed through the chromatographic column at least 20 times the volume of the column, and then washed in the reverse order.
2. Normal chromatographic column
Use n-hexane (HPLC grade), isopropanol (HPLC grade), dichloromethane (HPLC grade) and methanol (HPLC grade) as mobile phases respectively. Each mobile phase flows through the chromatographic column at least 20 times of the column volume (isopropanol has high viscosity, which can reduce the flow rate and avoid high pressure). Note that the order of using solvents should not be reversed. After washing with methanol, the reverse order should be followed Flush to n-hexane, all mobile phases must be strictly dehydrated.
3. Ion exchange chromatographic column
The ion exchange capacity of the chromatographic column will decrease when it is used in the buffer solution for a long time. The cation column can be regenerated by washing with dilute acid buffer solution, and the anion column can be regenerated by washing with dilute alkali buffer solution.
In addition, the solvent that can dissolve the pollutants in the column can be selected as the mobile phase for washing in the positive and reverse directions. If the column is reversed, it can be adjusted back, but the support in the column may collapse. If the column has to be reversed, the column should not be reversed.
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