Regeneration method of polymer chromatographic column in liq

Liquid chromatography is widely used in biomedical industry because of its high precision for polymer compounds with low boiling point. The polymer column used to separate biomolecules will be polluted if it is used for a long time, and then the column efficiency will be lower, which will affect the detection speed and sensitivity. Therefore, it is necessary to clean them regularly.
The chemical stability of polymerization data is usually measured by force. In general, the first use of nitric acid or sodium hydroxide solution to clean. Some cleaning methods of reversed phase polymerized chromatographic columns, such as polystyrene divinylbenzene beads and polymerized monomers, such as CIM rp-sdvb sheet and swift column, can withstand wide pH scale (usually ph11-13 or sometimes ph0-14). However, when the column is washed with strong organic solvent, it should be noted that the column will swell or shorten when washed with some organic solvents. Polymers with a crosslinking degree of more than 8-10% usually have good mechanical properties, with little shortening in aqueous solution and little swelling in organic solvents.
The method of regenerating polystyrene divinylbenzene whole column is as follows: using 0.1% 2-propanol to wash more than 10 column volumes at half the operating flow rate; 100% mobile phase B to wash more than 5 column volumes at half of the operating flow rate; using 100% mobile phase a to balance more than 10 volumes at the operating flow rate.
The protein residue on the silica gel bonded reversed phase column of cleaning liquid chromatography can be eliminated by washing sodium hydroxide, water, 20% ethanol solution and working buffer solution of 10 column volumes in sequence. If there are many hydrophobic proteins, the user should insert a 30% isopropanol or 70% ethanol process in the future.
If it is necessary to clean or inactivate microbial colonies, polystyrene divinylbenzene can be completely cleaned with 0.5-1.0 M sodium hydroxide. All columns should be washed with sodium hydroxide for at least one hour at room temperature.
It is necessary to use the column filled with traditional polymer matrix to separate some proteins with low solubility, such as membrane protein, structural protein and viral coat protein, etc. For example, 50% isopropanol solution containing 3M guanidine hydrochloride can only remove these proteins at 60 ℃.
The removal of synthetic peptides in solid phase resin can produce ab initio positive ions eliminated by anisole and phenylthiomethane. The elimination of positive ions produces large aromatic molecules, which contaminate the column during peptide purification. This contaminant has a very high retention on the C18 column and cannot be washed out by 100% methanol or acetonitrile. To clean this kind of column, the column must be reversed with 5 volumes of 100% isopropanol, three to five volumes of dichloromethane, then three to five volumes of isopropanol, Zui and then to the initial solvent system. The elution of aromatic impurities can be detected by 260 nm UV.
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