The History And Principles Of HPLC
HPLC is the abbreviation of high-performance liquid chromatography. It is some kind of chromatographic analysis technology and used to separate the mixtures to confirm and quantize the proportion of each composition. Basically, HPLC depends on the pump to pressure the samples and makes them go through the HPLC columns that are filled with adsorbing materials. As a result, each component of the samples can be separated. In general, HPLC is mostly applied in biological chemistry and analytical chemistry.
In the 1960s, as the gas chromatography technology had limitations on the analysis of the organics with high boiling points. In order to separate the macromolecular substances that are difficult to be gasified, such as protein and nucleic acid, the classical liquid chromatography was introduced to the theories and methods of gas chromatography. At the end of the 1960s, Kirkland and other scientists invented the first HPLC instrument in the world and thus opened the era of HPLC.
As a whole, HPLC uses stationary phases with smaller grain sizes to fill in the HPLC columns, increases the numbers of the column plates and drives the stationary phases with high pressure, which can make the separation finish within only hours or even dozens of minutes.
The Principles of Using HPLC Columns
In general, HPLC technology uses all kinds of interaction forces to separate the mixtures. These interaction forces are mostly some kind of non-covalent properties between the analytes and the HPLC columns. When using the HPLC columns, the liquid samples that are to be detected will be injected into the HPLC columns at different times. Through pressure moving in the stationary phases, as the interaction forces between different substances in the samples and the stationary phases are different, the substances will leave the HPLC columns in sequence. The laboratory workers will get different peak signals by using the detectors, and each peak point represents one kind of chemical compound. Finally, through analyzing and comparing the signals, we can judge the substances that are contained in the samples to be detected.
Furthermore, the chromatographic instrument that is designed with the liquid as the mobile phase is called liquid chromatography. In comparison, the liquid chromatography that is equipped with a high-pressure infusion pump, high-efficiency stationary phase, and the high-pressure sensitive detector is called high-performance liquid chromatography. There is a great variety of HPLC which are used in the laboratories. However, no matter what kind of HPLC, it has four basic parts: high-pressure infusion system, sample loading system, separation system, and detection system.
Good Separation Effect
Diol HPLC column is mainly used for the separation of peptides, proteins, and polar drug molecules. This column has ultra-pure fully porous spherical silica gel, which greatly improves the stability of chromatographic column separation and has good symmetrical peak shape and high column effect; besides, it ensures the inert, homogeneous and reproducible surface of silica gel with unique surface modification method. Moreover, Diol HPLC columns are mainly used for sample analysis, especially for mobile phase and sample filtration in chromatographic analysis, which has a good effect on protecting chromatographic column system and injection valve from contamination, and can be widely used in gravimetric analysis, microanalysis, colloid separation, and aseptic experiments.
Compared with the conventional HPLC column, the Diol HPLC column is a much more stable and reproducible orthophase column than the silica gel column and amino column. It is modified on the basis of pure silica gel, and some compounds can provide better selectivity and chromatographic peak shape. And it adopts unique bonding technology to ensure stable stationary phase structure and excellent batch and batch reproducibility.