High-performance liquid chromatography (HPLC) is a widely used analytical technique in which the mobile phase is a liquid. It is one of the oldest forms of chromatography, but through advancements in particle size, column pressure, and the introduction of gas chromatographic principles, it has evolved into a highly efficient method. HPLC uses high-pressure pumps, advanced stationary phases, and sensitive detectors to achieve fast analysis, high resolution, and automated operation. This technology has become essential in modern scientific research, especially in the analysis of complex mixtures.
In traditional Chinese medicine (TCM), where the composition is highly complex, conventional methods like thin-layer chromatography often fall short in terms of precision, accuracy, and reproducibility. HPLC, on the other hand, offers stability, reliability, and efficiency, making it the most important analytical tool for TCM research. It is commonly used to determine active compounds such as alkaloids, saponins, flavonoids, quinones, and coumarins in various herbal medicines.
Recent developments in HPLC have focused on improving sample preparation, detector selection, and column optimization. Techniques like solid-phase microextraction have enabled the detection of low-concentration components, while derivatization methods enhance sensitivity for substances with weak UV absorption. Ion-pairing reagents help improve retention of polar compounds, and coupling HPLC with mass spectrometry (LC-MS) has significantly advanced the field by providing accurate structural information.
The integration of HPLC with other technologies, such as fluorescence detection, evaporative light scattering detection, and nuclear magnetic resonance (NMR), has further expanded its application. These combinations allow for more comprehensive and accurate analysis of complex samples, especially in pharmaceutical and biomedical research.
Looking ahead, future trends in HPLC include reducing analysis time, increasing separation efficiency, and moving toward automation and intelligent systems. Multi-column techniques, gradient elution, and smart control systems are being explored to enhance performance and adaptability. Additionally, the combination of HPLC with advanced detection methods and molecular biology tools promises to revolutionize the study of TCM, supporting full-component analysis and deepening our understanding of herbal mechanisms.
Overall, HPLC continues to evolve, offering powerful solutions for analyzing traditional medicines and contributing to the advancement of modern pharmacology.
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