Short Description:
Preparative liquid chromatography: same structure as an analytical type, but with high pump flow, large injection volume, and preparative column; post-column fraction collector.
The purpose of the preparative HPLC column is to extract a certain component in a sample. If you want to obtain the component corresponding to a certain peak on the spectrum, you only need to extract the mobile phase from the peak start time to the end time.
Preparative scale liquid chromatography is used for the purification and isolation of molecules that are of value in the pharmaceutical, biopharma, and associated chemical industries. The production of highly pure compounds in these industries is increasingly important in terms of the chemical activity, toxicity and usefulness for screening in highly defined and often costly experimental processes.
Specifically designed for high performance preparative liquid chromatography (Prep HPLC).
Specifications:
Particle Size: 5 µm
Pore Size: 100 Å
Length: 250 mm
Inside Diameter: 10 mm
Installation of the HPLC column.
a. Unpack the column and confirm the HPLC column types, size, factory date, and the solvent stored in the column.
b. Unscrew the sealing plugs from the connectors at both ends of the column and return them to the box for spare use.
c. Connect the inlet end of the column to the outlet of the inlet valve through the connection tube in the direction of the mobile phase flow indicated on the column tube (if possible, it is recommended to use a guard column before the lc column); connect the outlet of the column to the detector. The connection tube is a stainless steel tube with an outer diameter of 1.57 mm and an inner diameter of 0.1 – 0.3 mm. Both ends of the connection tube have hollow screws and pressure rings for sealing. It is important to reduce the dead volume outside the column when taking over. After passing the hollow screw and pressure ring, try to tighten the hollow screw clockwise until it is not tightened, and then continue to tighten it clockwise with a wrench for 1/4-1/2 turn. Do not use too much force. If the column leaks after being pressurized by the mobile phase, continue to tighten it clockwise with a wrench for 1/4 turn until it does not leak.
FAQ
1. I remember that when I got a C18 column before, I would use methanol to slowly activate it from a small flow rate to a large flow rate. What is the purpose of doing this? Is it activated with a solvent first? Then use the sample? Also, the short peptides are always flushed out. How to solve it? It is with the solvent peak, or before the solvent peak.
– The C18 column will be activated slowly with methanol from a small flow rate to a large flow rate because the time for the chromatographic column to reach the user is different. During the storage and transportation process, the storage liquid may be a little volatilized. Wetting the stationary phase to make the bonded phase stretch well. After balancing the system with solvent, you can take multiple injections or increase the injection volume to obtain reproducible chromatograms faster. In this way, the sample will have The active sites in the chromatographic column saturated, and there will be no abnormal chromatographic phenomena.
2. The C18 column used in the national standard is used to measure the spiciness of pepper essence, and Tween series emulsifiers are added to the pepper essence to make water-soluble capsicum essence. Does the emulsifier affect the C18 column?
– Emulsifiers are similar to ion-pair reagents, with polar and non-polar ends, which will definitely affect the C18 column. However, if capsaicin is a polar substance, the presence of an emulsifier can improve its retention on a C18 column.
3. What is the protective solvent for a new liquid chromatographic column? What is the best way to balance and clean, and how long should it take to balance?
– Depending on the type of chromatographic column, the storage solvent will also be different, depending on the instructions in the manual. For reversed-phase columns, methanol (acetonitrile) is generally used for storage, and methanol/water with a methanol concentration of about 80% is also used for storage. Generally, 10-20 column volumes are enough to equilibrate with the mobile phase. (Note: The column volume is not equal to the volume of the empty column tube. The empty column tube volume of a 4.6×250mm chromatographic column is 4.15mL, while the column volume is only about 2.5mL)
FAQ
1. I remember that when I got a C18 column before, I would use methanol to slowly activate it from a small flow rate to a large flow rate. What is the purpose of doing this? Is it activated with a solvent first? Then use the sample? Also, the short peptides are always flushed out. How to solve it? It is with the solvent peak, or before the solvent peak.
– The C18 column will be activated slowly with methanol from a small flow rate to a large flow rate because the time for the chromatographic column to reach the user is different. During the storage and transportation process, the storage liquid may be a little volatilized. Wetting the stationary phase to make the bonded phase stretch well. After balancing the system with solvent, you can take multiple injections or increase the injection volume to obtain reproducible chromatograms faster. In this way, the sample will have The active sites in the chromatographic column saturated, and there will be no abnormal chromatographic phenomena.
2. The C18 column used in the national standard is used to measure the spiciness of pepper essence, and Tween series emulsifiers are added to the pepper essence to make water-soluble capsicum essence. Does the emulsifier affect the C18 column?
– Emulsifiers are similar to ion-pair reagents, with polar and non-polar ends, which will definitely affect the C18 column. However, if capsaicin is a polar substance, the presence of an emulsifier can improve its retention on a C18 column.
3. What is the protective solvent for a new liquid chromatographic column? What is the best way to balance and clean, and how long should it take to balance?
– Depending on the type of chromatographic column, the storage solvent will also be different, depending on the instructions in the manual. For reversed-phase columns, methanol (acetonitrile) is generally used for storage, and methanol/water with a methanol concentration of about 80% is also used for storage. Generally, 10-20 column volumes are enough to equilibrate with the mobile phase. (Note: The column volume is not equal to the volume of the empty column tube. The empty column tube volume of a 4.6×250mm chromatographic column is 4.15mL, while the column volume is only about 2.5mL)
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