Balance and use of the hottest spectral column

2022-09-21
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Balance and use of chromatographic column

the reverse chromatographic column is stored in methanol (or acetonitrile)/water after factory test. Please make sure that the mobile phase you use is miscible with methanol (or acetonitrile)/water. Because the chromatographic column may dry out during transportation, you should use multiple volumes of methanol or acetonitrile to balance the chromatographic column before analyzing the sample with the mobile phase

if the mobile phase you use contains buffer salt, you should pay attention to the transition with the mobile phase with high water content (about 95%), and the same is true when washing the column after the sample Inner Mongolia (Naiman) is analyzed by an nonferrous materials Co., Ltd. In the past, the traditional method used pure water flushing. For non-polar columns, such as C8 or C18, because pure water cannot infiltrate and fill, if the oil hole of the buffer is blocked by the surface of the material and the carbon chain is washed down, the column efficiency will be reduced

in addition, when analyzing samples, because pure water cannot infiltrate and form a water film on the surface of the filler, the samples are not retained and cannot be separated, and the reproducibility is poor, so for general C18 or C8, the organic solvent cannot be less than 5%. Silica gel column or polar chromatographic column is stored in n-heptane after factory test. If the chromatographic column needs to use a mobile phase containing water or methyl alcohol or acetonitrile, please balance it with ethanol or isopropanol before using the mobile phase

how to balance the chromatographic column:

during the balance process, slowly increase the flow rate and balance the chromatographic column with mobile phase until a stable baseline is obtained (if the concentration of buffer salt or ion pair reagent is low, it takes a long time to balance)

regeneration of chromatographic column:

recommended solvent volume for flushing

column specification column volume solvent volume

150 × 4.6 2.50ml 50ml

250 × 4.6 5.0ml 100ml

250 × 10 20ml 400ml

please choose your regeneration method for expanding its application field according to the following table:

regeneration of polar stationary phase (such as Si, NH2, diol chromatographic filler): n-heptane chloroform ethyl acetate acetone ethanol water

regeneration of non-polar stationary phase (such as reverse filler C18, C8, etc.): water acetonitrile chloroform (or isopropanol) - acetonitrile water

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