High Performance Liquid Chromatograph (HPLC) system is an important branch of chromatography and is an indispensable separation and analysis technology in the fields of modern chemistry, medicine, industry, agriculture, commodity inspection and judicial inspection. It uses liquid as the mobile phase and uses a high-pressure infusion system to pump a single solvent with different polarities or mixed solvents, buffers, etc. into a chromatographic column equipped with high-efficiency stationary phase. After the components in the column are separated, they enter the detector for detection, thereby achieving efficient, fast and sensitive analysis of the sample.
Structural components
The high-performance liquid chromatography system mainly includes liquid storage tanks, high-pressure infusion pumps, samplers, chromatographic columns, detectors, recorders, and data processing devices. Among them, the high-pressure infusion pump is one of the core components of the entire system, which is responsible for sending the mobile phase into the chromatographic column with stable pressure and flow. The chromatographic column is a key component of separation, filled with a stationary phase with extremely fine particles. The partition coefficients of different compounds between the stationary phase and the mobile phase are different, thereby achieving separation. The detector converts the separated components into electrical signals, and outputs the graph and results through the recorder and data processing device.
How it works
The working principle of the high-performance liquid chromatography system is based on the separation principle of chromatography, that is, when each component dissolved in the mobile phase passes through the stationary phase, it has different sizes and strengths of the interaction with the stationary phase (adsorption, distribution, exclusion, and affinity), and the residence time in the stationary phase is different, so it flows out of the stationary phase one after another. Specifically, the high-pressure pump penetrates the mobile phase in the liquid storage tank into the system, and the sample solution enters the mobile phase through the injector, and is loaded into the chromatographic column as the mobile phase flows. Because the distribution coefficients of the components in the sample solution are different in the mobile phase and the stationary phase, when the two phases are moved relative to each other, after continuous adsorption-desorption distribution process, the distance between the mixed components is gradually opened, and finally flows out of the column in sequence as separate individual components. The detector converts the sample concentration into an electrical signal and transmits it to the recorder to print out the sample data in the form of a graph.
Advantages and applications
The high performance liquid chromatography system has the characteristics of high separation efficiency, fast analysis speed, good detection sensitivity, and the ability to analyze and separate thermally unstable physiologically active substances with high boiling points and cannot be vaporized. Its application range is wide and is almost not limited by the volatility and thermal stability of the analytical objects. Almost all compounds, including high boiling point, polarity, ionic compounds and macromolecular substances, can be analyzed and determined by high performance liquid chromatography. In the fields of pharmacy, traditional Chinese medicine and traditional Chinese medicine, food science and technology, high performance liquid chromatography systems are widely used in drug analysis, Chinese medicine fingerprint research, pesticide residue detection, food nutritional component analysis, etc.
Development trends
With the rapid development of modern biotechnology and pharmaceutical industry, high-performance liquid chromatography systems are also constantly developing and improving. The current development trend is to tilt towards biochemical and drug analysis and preparation, and to develop more efficient, sensitive and rapid separation and analysis methods. At the same time, in response to the problems of high consumption and toxicity of mobile phases, environmentally friendly and sustainable alternatives are also actively exploring.
To sum up, high performance liquid chromatography systems play an increasingly important role in the field of modern science and technology with their unique separation and analysis capabilities and wide application prospects.
