Automated Flash Column Chromatography
2.Chromatographic Column (Rotation Type)
3.Chromatographic Column (Manual)
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Description
Technical Parameters
Automated Flash Column Chromatography is an efficient mixture separation technique, particularly suitable for rapid separation and purification of complex mixtures. Its basic principle is similar to column chromatograph and thin-layer chromatograph, both of which are based on the difference in distribution coefficients of different substances between the stationary phase and the mobile phase for separation. However, automatic rapid column chromatograph significantly improves separation efficiency and speed by using automated equipment and compressed air to push the solvent.
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Parameter
Principles of Physics and Operating Procedures
1. Principles of Physics
(1) Distribution coefficient: The distribution coefficient of a substance between the stationary phase and the mobile phase is a key factor determining its separation rate. The larger the distribution coefficient, the longer the retention time of the substance on the stationary phase; On the contrary, the smaller the distribution coefficient, the faster the material moves with the mobile phase.
(2) Adsorption capacity: The adsorption capacity of the stationary phase (such as silica gel) on substances can also affect the separation efficiency. Substances with strong adsorption capacity have a longer residence time on the stationary phase, while substances with weak adsorption capacity are more likely to move with the mobile phase.
(3) Solubility: The solubility of a substance in the mobile phase also affects its separation rate. Substances with high solubility are more likely to move with the mobile phase, while substances with low solubility may remain on the stationary phase for a longer period of time.
2. Operation process
(1) Sample preparation: Firstly, it is necessary to determine the dryness level of the mixture to be separated and weigh it. Then, based on the results of thin-layer chromatograph, select an appropriate solvent system to keep the Rf value within an appropriate range (usually 0.2~0.3). For complex mixtures, gradient elution may be necessary.
(2) Sample column loading: Depending on the properties of the sample, the net sample method, solution method, or silica gel adsorption method can be selected to load the sample onto the colum. The net sample method is suitable for non viscous oily substances, the solution method is suitable for liquid and solid samples, and the silica gel adsorption method is suitable for some liquid and all solid samples.
(3) Silicone filling: Choose the appropriate ratio of silicone and compound, as well as the appropriate size of the separation colum. Then, fill the fume hood with a separation colum, ensuring that the colum is vertical and avoiding loss of silicone gel. Add sand as a protective layer on top of the silicone to prevent damage to the silicone layer.
(4) Elution and collection: Add eluent to the separation colum to begin the elution process. Maintain an appropriate flow rate and regularly replace the collection tubes. Meanwhile, thin-layer chromatograph can be used to track the separation process to ensure the collection of pure samples.
(5) Gradient elution: For complex mixtures, gradient elution may be necessary. Gradually changing the polarity of the eluent to elute compounds of different polarities. During the gradient elution process, it is necessary to slowly increase the polarity of the solvent to avoid silica gel splitting and a decrease in separation efficiency.
Application area
Automated Flash Column Chromatography technology has been widely used in multiple fields due to its high efficiency, speed, and ease of automation. Here are some of the main application areas:
(1) Natural product extraction and separation: Automatic rapid colum chromatograph technology can be used to extract and separate biologically active compounds from natural products. These compounds have broad application prospects in fields such as drug development, food industry, and cosmetics.
(2) Drug synthesis and purification: In the process of drug synthesis, automatic rapid colum chromatograpy technology can be used for the purification of synthesized products and the removal of impurities. This helps to improve the purity and quality of drugs, thereby meeting the requirements of drug regulatory agencies.
(3) Environmental pollutant analysis: Automatic rapid colum chromatograph technology can be used for the analysis and detection of pollutants in environmental samples. By selecting appropriate stationary and mobile phase conditions, effective separation and enrichment of trace pollutants in environmental samples can be achieved, providing scientific basis for environmental pollution control.
(4) Food safety testing: In the field of food safety testing, automatic rapid colum chromatograph technology can be used for the detection of harmful substances such as additives and pesticide residues in food. This helps ensure the safety and quality of food, and safeguard the health rights of consumers.
(5) Biochemistry research:
In the field of biochemistry research, automatic rapid colum chromatograph technology can be used for the separation and purification of biomolecules such as proteins, nucleic acids, etc. These biomolecules play important roles in life science research, disease diagnosis, and treatment.
The physical phenomena and factors involved
Automated Flash Column Chromatography technology itself does not have fixed physical properties, it involves various physical phenomena and factors during operation, which are crucial for understanding and optimizing the technology.
1. Physical properties of mobile phase
(1) The choice of solvent:
The physical properties of the solvent, such as polarity, viscosity, density, etc., have a significant impact on the separation efficiency. Choosing a suitable solvent system can ensure that the substance to be separated has an appropriate distribution coefficient between the stationary phase and the mobile phase.
(2) Control of flow rate:
Flow rate is an important factor affecting separation speed. A faster flow rate can shorten the separation time, but it may also lead to a decrease in separation efficiency. Therefore, it is necessary to adjust the flow rate according to the actual situation to achieve the best separation effect.
2. Physical properties of stationary phase
(1) The properties of silicone:
Silicone is one of the commonly used stationary phase materials. The physical properties such as particle size, porosity, and specific surface area can all affect the separation efficiency. Silica gel with smaller particle size has a larger specific surface area and porosity, which can provide better separation efficiency; However, particles that are too small may also cause colum blockage and a decrease in separation speed.
(2) Stability of stationary phase:
The stability of stationary phase is crucial for long-term operation and repeated experiments. A stable stationary phase can ensure the accuracy and reliability of the separation results.
3. The physical structure of the separation column
(1) The size and shape of the column:
The size and shape of the colum will affect the separation effect and flow rate. A shorter colum can shorten the separation time, but it may also lead to a decrease in separation efficiency; Longer colums may provide better separation efficiency, but the separation time will also be correspondingly prolonged. The shape of the colum also affects the separation effect, and cylindrical colums are usually chosen to ensure uniform distribution of the mobile phase.
(2) The tightness of the packing:
The tightness of the packing will affect the flow rate and separation effect of the mobile phase. Overly tight packing may lead to a decrease in flow rate and separation efficiency; Overly loose packing may lead to colum blockage and incomplete separation.
4. The influence of temperature and pressure
(1) Temperature control:
Temperature is one of the important factors affecting separation efficiency. Appropriate temperature can ensure optimal volatility of solvents and solubility of substances, thereby improving separation efficiency. Excessive temperature may lead to rapid solvent evaporation and substance degradation; However, excessively low temperatures may lead to a decrease in solvent solidification and separation rate.
(2) Pressure control:
In automatic rapid colum chromatoraphy, compressed air is usually used to push the solvent through the colum. Therefore, pressure control is crucial for ensuring the stability of flow rate and the accuracy of separation effect.
Application and Optimization of Physical Phenomena
(1) Diffusion and convection:
In the separation process, diffusion and convection are two important physical phenomena. Diffusion refers to the process of transferring substances from high concentration areas to low concentration areas under the action of concentration gradients; Convection refers to the transfer of matter caused by fluid motion. In automatic rapid colum chromatograhy, diffusion and convection work together to affect the separation speed and efficiency of substances. Therefore, it is necessary to improve diffusion efficiency and convection velocity by optimizing operating conditions and colum structure, in order to achieve better separation effects.
(2) Chromatograph effect:
Chromatograph effect refers to the process of multiple adsorption and desorption of substances between the stationary phase and the mobile phase. This effect can increase the retention time of substances in the colum, thereby improving the separation efficiency. However, excessive chromatographic effects may also lead to a decrease in separation speed and colum blockage. Therefore, automated flash column chromatography is necessary to balance the relationship between chromatograph effect and separation speed during the operation process to achieve the best separation effect.
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