Progress in the application of high performance liquid chromatography in traditional Chinese medicine research

One of the old zui in chromatography, liquid chromatography refers to the chromatographic technique in which the mobile phase is liquid. However, by improving the particle size and column pressure of the filler, the classical liquid chromatography is based on the theory of gas chromatography. The technology uses a high-pressure infusion pump, high-efficiency stationary phase and high-sensitivity detector to achieve fast analysis, high separation efficiency and automatic operation. This chromatographic technique is called high performance liquid chromatography (HPLC). Complex, conventional thin-layer chromatography and other methods can not meet the needs of modern Chinese medicine because of its precision, accuracy, sensitivity and reproducibility. High-performance liquid chromatography is a stable, reliable and efficient Chinese medicine research. The important analytical method of Zui. At present, high performance liquid chromatography has been widely used in the determination of active ingredients of various traditional Chinese medicines such as alkaloids, saponins, flavonoids, quinones and coumarins. Many, because of the advantages of HPLC and GC in high performance liquid chromatography, regardless of efficiency and selectivity The level of analysis has reached or exceeded. In recent years, the lack of high performance liquid chromatography has been improved, and this technology has been improved.
Through the study of different compatibility and decoction time from single-flavor to compound, the general trend of traditional Chinese medicine research is full-component analysis. Only when we discover the changes of chemical composition in traditional Chinese medicine, we find the organic mechanism between Chinese medicines. There are many kinds of traditional Chinese medicines, and the properties of various ingredients are popular in all polar segments and acid-base ranges. The simple way to achieve multi-component analysis is to use a mild mobile phase on a long enough column to elute for a long enough time. But this is contrary to the simplicity and speed of modern analytical requirements. Through a large number of applied studies, high-performance capillary electrophoresis has great advantages in the analysis of traditional Chinese medicine components, especially in the analysis of high-polarity chemical components, and has shown a high ability in analyzing a large number of compound preparations. Since the capillary tube is almost free of bed contamination that often occurs in high performance liquid chromatography, and the used capillary column is rinsed in a short time, the second sample can be analyzed, fast and efficient, and resolution. Very high. Emerging capillary electrochromatography is a new type of electro-separation microcolumn liquid chromatography technology that combines the advantages of high performance liquid chromatography and capillary electrophoresis. It moves a variety of filler particles from high performance liquid chromatography into a capillary tube between the sample and the stationary phase. The interaction acts as a separation mechanism to electroosmotic flow as a mobile phase driving force for the chromatographic process. Zui is close, some advanced testing instruments have been successfully used in high performance liquid chromatography, which makes the application of high performance liquid chromatography more widely, and makes full use of the advantages of high efficiency, fastness, good selectivity and high sensitivity to establish a more systematic component analysis. method. Combined with mass spectrometry, gradient elution, column switching technology, advanced detection technology, and molecular biology, modern molecular pharmacology, it will play a significant role in the analysis of traditional Chinese medicine.

1 Recent developments in high performance liquid chromatography

Mainly consider the pre-treatment of the sample and the selection of the analytical column and detector. The pretreatment of the sample, the application of high performance liquid chromatography in the analysis of the drug. The solid phase extraction which has been developed recently has made many low-content components refined and purified, which is suitable for the determination of high performance liquid chromatography, and Sun Xinguo uses countercurrent extraction. The content of ligustrazine has achieved good results. Some of the traditional Chinese medicines have weak UV absorption or no characteristic UV absorption. They are directly determined by high performance liquid chromatography. Their sensitivity and resolution are not satisfactory. These components can be accurately determined by pre-column or post-column derivatization. come out. For the polar and fat-soluble substances, the YWGCl8 column is not easy to retain. The sodium dodecyl sulfate is used as the ion-pairing reagent to reduce the polarity and prolong the storage time on the column, so that better separation is achieved. The two powerful analytical techniques of liquid chromatography and mass spectrometry are linked together online. After 30 years of development, it has become a more mature analytical tool, but from the beginning of the formation there is a problem: flowing from the liquid chromatography At the time of mass spectrometry, the change of mobile phase, the composition of solvent, and the problem of high temperature and high pressure ionization restrict the development of this combination technology. The atmospheric pressure ionization interface has the dual functions of removing solvent and ionization, and the introduction of this technology makes it possible in various fields. A wide range of applications. Electrospray ion source is a soft ionization technique, generally only generated (M+H+ and (MH- two molecular ion peaks, selective monitoring (mz190 negative molecular ion peak, with high sensitivity, accuracy, specificity) It satisfies the needs of low-concentration drug research. The three-dimensional high performance liquid chromatography method studied by Zhang Li et al. can simultaneously determine two indicators of puerarin and ferulic acid. It is proved by experiments that if the appropriate column temperature and other chromatographic conditions are selected, Ethanol is used as the mobile phase of reversed-phase high performance liquid chromatography to analyze the active ingredients in traditional Chinese medicines and proprietary Chinese medicines. It is safe and accurate. It is difficult to detect common structures with similar structures. High performance liquid chromatography-electrochemical method can effectively determine The content of baicalin and baicalein in the berberine powder is only one group. The sample is separated by chromatography, and the fluorescence intensity is measured by fluorescence spectrophotometer. The influencing factors are many, the determination is complicated, and the improved HPLC is performed. Fluorescence can be obtained without derivatization and collection of the isolates, only by chemical treatment, and can be directly injected after concentration. The method for determining the content of hypericin in Hypericum perforatum L. has also achieved a good knot. High performance liquid chromatography-differential refractive index measurement of Astragalus membranaceus oral liquid in the content of astragaloside has also achieved satisfactory results. For only the UV end absorption, the sensitivity is low with UV detection, the baseline is easy to drift, differential refractive index detection is easy Under the interference of external conditions, the evaporative scatter detector can overcome the above lack, the response value is only related to the sample quality, and its signal is proportional to the mass. The different compounds have the same quality when the quality is the same. The evaporative light scattering detection method is based on non-volatile The scattering level of the sample molecules to light is directly proportional to its mass, regardless of the nature of the groups it contains. By selecting the appropriate detector parameters, the mobile phase and solvent can be completely vaporized without generating a signal, and the individual components in the sample. The presence of non-volatile particles scatters light to be detected. Therefore, the evaporative light scattering detector can be used for simultaneous separation and analysis of multiple components containing different groups. Compared with ultraviolet, fluorescence, etc., evaporative light scattering The detection method has approximately the same response factor for different substances.

It is beneficial to the accurate determination of mixtures in different proportions. High performance liquid chromatography-evaporative light scattering test for the determination of terpene lactone content in Ginkgo biloba leaves, ginseng in red ginseng and Yujing capsules, thus no low concentration, high response or high concentration, low The phenomenon of response.

The contents of saponin Rg1 and Re and the content of gambogic acid in garcinia were all very good.

2 Research trends in high performance liquid chromatography

Improve separation efficiency. Using advanced detection instruments and methods, 2.1 shortening the analysis time. Adjusting or changing the mobile phase and stationary phase, gradient elution and column switching technology are expected to solve this problem. Gradient-eluting high-performance liquid chromatography, which can analyze samples with a wide range of polarities, has great advantages over isocratic elution, but it is somewhat inconvenient for traditional Chinese medicine samples with more complex components and wider polar ranges. Multi-column high performance liquid chromatography is also known as multidimensional high performance liquid chromatography. In addition to the advantages of changing the concentration of the mobile phase with gradient elution, the type of stationary phase, the degree of bonding, the particle size, the length of the column, the diameter of the column, and the type and concentration of the mobile phase can be changed.

LC-MS-MS method for the determination of HIV-1 protease in plasma, 2.2 further developed into automation, intelligence and combination technology. The combination of liquid chromatography and mass spectrometry has become the first choice for the determination of drugs and metabolites in low-concentration biopharmaceuticals. It is accurate and efficient. The endogenous components and other drugs in the plasma do not interfere with the determination, saving data and saving time. . Has been applied to the analysis of drugs in body fluids, plasma, serum. The analysis of cholic acid in Chinese herbal compound injection "Qing Kai Ling" was carried out by liquid chromatography-mass spectrometry and mass spectrometry. High performance liquid chromatography-nuclear magnetic resonance (NMR) has a very good role in drug analysis. The newly proposed intelligent multi-column HPLC system utilizes the modular separation performance of the switching technique to switch the sample into blocks of different properties and elute with a suitable mobile phase. Intelligent control is used throughout the process.

3 Application prospect of high performance liquid chromatography in traditional Chinese medicine analysis

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