Medicinal Plant Research 2024, Vol.14, No.1, 45-56 http://hortherbpublisher.com/index.php/mpr 48 4 Analytical Methods for Chemical Characterization 4.1 Extraction techniques 4.1.1 Solvent extraction Solvent extraction is a widely used method for isolating bioactive compounds from plant materials. In the study by (Chen et al., 2021), various solvents such as 95% ethanol, ethyl acetate, n-hexane, and distilled water were employed to extract different components from Chrysanthemum morifolium Ramat. The ethanolic extract demonstrated the highest levels of total flavonoids, polyphenols, and chlorogenic acids, indicating its efficacy in extracting these bioactive compounds. Similarly (Li et al., 2019b), utilized hot water and 75% methanol for extracting chemical compositions from commercial chrysanthemum teas, highlighting the versatility of solvent extraction in different solvent systems. 4.1.2 Supercritical fluid extraction Supercritical fluid extraction (SFE) is an advanced technique that uses supercritical fluids, typically CO2, to extract bioactive compounds. Although specific studies on SFE for Chrysanthemum morifoliumwere not provided in the data, this method is known for its efficiency and environmental friendliness. SFE can selectively extract compounds based on their solubility in supercritical CO2, which can be fine-tuned by adjusting temperature and pressure. 4.2 Chromatographic methods 4.2.1 High-performance liquid chromatography (HPLC) High-Performance Liquid Chromatography (HPLC) is a powerful analytical technique used to separate, identify, and quantify components in a mixture. In the research by Chen et al. (2020), HPLC was employed to construct a spectral tag library for identifying and quantifying various bioactive compounds in Chrysanthemum morifolium flowers. This method allowed for the precise determination of flavonoids, caffeoylquinic acids, and other compounds, demonstrating its utility in chemical characterization. 4.2.2 Gas chromatography-mass spectrometry (GC-MS) Gas Chromatography-Mass Spectrometry (GC-MS) combines the features of gas chromatography and mass spectrometry to identify different substances within a test sample. The study by (Youssef et al., 2020) utilized GC-MS to analyze the essential oils of Chrysanthemum morifolium, identifying camphor as a major constituent. Additionally (Sayed, 2023), employed GC-MS to discover bioactive substances such as lupeol and α-amyrin in the acetone extracts of Chrysanthemum morifolium, showcasing the method's capability in detailed chemical profiling. 4.3 Spectroscopic methods 4.3.1 Nuclear magnetic resonance (NMR) Nuclear Magnetic Resonance (NMR) spectroscopy is a technique used to determine the structure of organic compounds by observing the behavior of nuclei in a magnetic field. In the study by (Loh et al., 2021), a proton NMR-based metabolomics approach was used to characterize xanthine oxidase inhibitors in Chrysanthemum morifolium. This method provided detailed insights into the metabolite profile and helped identify key compounds responsible for the observed bioactivity. 4.3.2 Infrared (IR) spectroscopy Infrared (IR) spectroscopy is used to identify and study chemicals through their infrared spectra. Although specific studies on IR spectroscopy for Chrysanthemum morifoliumwere not provided in the data, this technique is commonly used to identify functional groups and characterize molecular structures in plant extracts. IR spectroscopy can complement other analytical methods by providing additional information on the chemical composition of the extracts. In summary, a combination of solvent extraction, chromatographic, and spectroscopic methods provides a comprehensive approach to the chemical characterization of Chrysanthemum morifolium. These techniques enable
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