Detailed explanation of performance parameters of liquid Chr
This paper mainly introduces the related parameters of liquid chromatography column, as follows.1. Surface area
The sum of the outer and inner pore surfaces of the particle, expressed in m2 / gram.
High surface area has strong retention capacity, column capacity and resolution for multicomponent sample separation. The packing with low surface area can reach equilibrium rapidly, which is especially important for gradient leaching.
2. Aperture
The average size of the pore or cavity of the particle, ranging from 60-10000 Å.
The results show that the retention time of solute macromolecules on the surface of packing can be prolonged by macroporous filler particles, which can achieve full separation and improve the peak shape; the pore size of 80 ° is selected for sample MW ≤ 4000, and 300 ° for sample MW > 4000
3. Carbon coverage
The effect on chromatographic separation, the amount of bonding phase connected with the matrix material.
High carbon coverage can improve the resolution and the analysis time is long. Low carbon coverage reduces operation time.
4. Sealing end
The effect of end capping on chromatographic separation, the use of shorter alkane chains to bond free silica hydroxyl groups (secondary bonding)
For polar samples, there is a significant difference in selectivity between the unsealed and the capped columns.
5. Particle size
5 um is the industry standard, and 7 um is the preparation column.
Extended reading: how to flush a liquid chromatography column
1) For the washing of reversed phase chromatographic column
Wash the chromatographic column (analytical column) with at least 25ml of the following solvents: mobile phase without buffer salt, 100% methanol, 100% acetonitrile, 75% acetonitrile + 25% isopropanol, 100% isopropanol, 100% dichloromethane and 100% hexane.
Note that if the column is washed with hexane or dichloromethane, the column must be washed with isopropanol before the reverse phase mobile phase is reused.
2) For washing of normal chromatographic column
Wash the chromatographic column (analytical column) with at least 50ml of the following solvents: 50% methanol + 50% chloroform, 100% ethyl acetate.
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