Jiangsu Manrui New Materials Co., Ltd , https://www.manruiwire.com
1. First, make sure the column and instrument fittings and piping match. In order to reduce the dead volume, the inner diameter of the connecting line between the injection valve, column, and detector should be as small as possible, while controlling the length of the connecting line between the injector, the column, and the detector. Before installing the column, verify that the solvent in the flow system is normal. For the analysis of more complex samples, it is recommended to install guard columns.
2. In order to achieve the best connection between the column and the instrumentation system, nut and cone fittings that match the column interface should be used as much as possible. If the original fittings are long-term compatible with other types of columns, it is recommended to connect new chromatography. Check the match before the column to avoid damage to the column or leakage due to column mismatch.
3. Use universal joints of PEEK material. Hand-tightening does not require special wrenches. The use pressure is 5000psi; the use temperature must not exceed 100°C.
Mobile phase
1. Before using the sample, use at least 20 column volumes of mobile phase to fully equilibrate the column. The mobile phase must use chromatographic grade solvents. If the aqueous phase buffer is used, it should be formulated daily to keep it fresh from bacteria.
2. The mobile phase needs to be filtered by a microporous membrane before use to eliminate the damage of the particles in the mobile phase to the chromatographic system and the column. After mixing the buffer with other mobile phases, it should be re-filtered to avoid new solubility due to solubility changes. The C18 column should not be rinsed with pure water as a mobile phase to avoid column performance deterioration (flushing the column with 5% organic solvent can also achieve the buffer salt cleaning effect. It can also make the column easier to equilibrate).
3. The mobile phase should be used after degassing to avoid abnormal operation of the pump and detector caused by bubbles. If the mobile phase used during the test is significantly different from the mobile phase used for column preservation, the over-distribution should be used for the balance. Avoid damage to the column and instrumentation system caused by an excessive increase in column pressure due to sudden changes in the mobile phase or crystallization of the mobile phase buffer salt. Normal phase columns require longer equilibration times than reversed phase columns.
Sample Preparation
1. The sample should be dissolved as much as possible in a solvent that is compatible with the mobile phase. Unless otherwise specified, the resolution of the sample column may be reduced if a strong solvent is used.
2. The sample solution should be filtered beforehand by using a needle filter before sampling. Changing the composition of the mobile phase frequently will accelerate the decrease of efficiency.
3. The column is filled with high-pressure homogenizer and can withstand higher pressures. In order to obtain the best separation effect, please do not exceed 200kgf when using, and avoid sudden increase or change in pressure, otherwise it will cause damage to the silica gel packing and reduce the service life of the column. The maximum operating temperature shall not exceed 60°C unless otherwise specified.
Save operation
1. If the mobile phase contains acid or inorganic salts, rinse with deionized water (20 column volumes) first. Then save the column with 100% acetonitrile or methanol. Finally sealed with the column's joints and stored in a stable environment.
2. Avoid direct mechanical shock or drop of the column to avoid degradation of column performance.
Column regeneration
Rinse the column in the following order with the equivalent of 20 column volumes of solvent. It is recommended that the column be flushed in the direction of the arrow so that it does not recoil as much as possible. The reference solvent sequence used for flushing is water, methanol, chloroform, isopropanol.