Three points for attention in the use of liquid chromatograp
OneInstallation of chromatographic column
Avoid strong collision and vibration when using chromatographic column. The chromatographic columns produced by different manufacturers have different interface sizes. If the type of column head and the type of stainless steel capillary joint do not match, there will be leakage or excessive dead volume. The depth of the joint is longer than that of the column head, so it is not easy to tighten and leak liquid; the depth of the joint is shorter than that of the column head, and there is a gap in the column head, resulting in the birth and death volume, broadening the spectrum band and tailing the peak.
Two
Equilibrium of chromatographic column
The equilibrium of chromatographic column refers to the complete analysis program, running several times of test, etc. until the peak shape, retention time and peak area are stable, the balance is completed. In addition, when using buffer salt, the buffer salt should be introduced into the chromatographic column after water transition to prevent salting out and damaging the chromatographic column.
Three
Preservation of chromatographic column
It is best to follow the retention method in the column instructions. In practical application, it can also be used as follows: it is better to keep the mobile phase used for short-term retention, which can reduce the balance time of the next use. In general, it is recommended to use 80% methanol or acetonitrile for a long time. The pure organic phase can reduce the hydrolysis of the bonded phase to the maximum extent, but the pure methanol (acetonitrile) will elute the hydrolyzed and temporarily adsorbed bonded phase in the column, which will accelerate the loss process of the stationary phase and shorten the service life of the stationary phase. In addition, when the pure organic phase is preserved, the solvent is easy to volatilize, resulting in the drying of the column bed and easy damage to the chromatographic column. In general, the positive phase column should be kept in the used mobile phase.
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