High-performance liquid chromatography (HPLC) is a well-established analytical technique in which the mobile phase is a liquid. It has evolved significantly from classical liquid chromatography, with improvements in particle size, column pressure, and the introduction of gas chromatographic principles. By incorporating high-pressure pumps, advanced stationary phases, and sensitive detectors, HPLC enables fast analysis, high separation efficiency, and automated operation. This technology has become the most critical method for analyzing traditional Chinese medicine (TCM), due to its stability, reliability, and efficiency. TCM contains complex chemical compositions, and conventional techniques like thin-layer chromatography often fail to meet modern requirements in terms of precision, accuracy, sensitivity, and reproducibility. HPLC has been widely used to determine active components such as alkaloids, saponins, flavonoids, quinones, and coumarins in various TCMs.
In recent years, research on the application of HPLC in TCM monitoring has grown rapidly. The integration of HPLC with other techniques, such as mass spectrometry, has enhanced its performance in terms of efficiency, selectivity, and analysis. Despite some limitations, such as solvent changes and ionization issues when coupling with mass spectrometry, advances like atmospheric pressure ionization interfaces have improved the applicability of these combined systems. Electrospray ionization, for example, offers high sensitivity and specificity, making it ideal for low-concentration drug analysis.
Recent developments in HPLC include improved sample pretreatment methods, such as solid-phase microextraction, which helps isolate low-concentration compounds. Derivatization techniques are also widely used for substances with weak or no UV absorption. Ion-pairing reagents, like sodium dodecyl sulfate, help separate polar and lipophilic compounds more effectively. Additionally, the combination of HPLC with fluorescence detection, evaporative light scattering detection, and refractive index detection has expanded the range of applications, especially for compounds that are difficult to detect using traditional methods.
Research trends in HPLC focus on reducing analysis time and improving separation efficiency through gradient elution, column switching, and multi-dimensional HPLC. Automation and intelligent control systems are also being developed to enhance accuracy and reduce manual intervention. The integration of HPLC with mass spectrometry and nuclear magnetic resonance (NMR) has opened new possibilities in drug analysis and metabolite identification.
Looking ahead, HPLC will play an even greater role in the full-component analysis of TCM. With the development of capillary electrophoresis and capillary electrochromatography, the ability to analyze complex mixtures has increased. These emerging techniques offer faster, more efficient, and higher-resolution separations, complementing traditional HPLC methods. Combined with advanced detection technologies and molecular biology, HPLC will continue to advance the understanding and quality control of traditional Chinese medicine.
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