| Supercritical Fluid Chromatography(SFC) is a chromatographic technique which is developed in 1980 s and based on Supercritical Fluid(SCF) as mobile phase. As a new type of chromatographic separation technology, the SFC has a lot of advantages, such as efficient, fast, low dosage of organic solvent, simple post-processing, less organic solvent residues in products, and is according to the new concept of green environmental protection. Therefore, SFC is favored by many scholars both at home and abroad. In recent years, its application areas are more and more widely, such as in the areas of separation of chiral drugs, separation and purification of Chinese medicines and the natural products, and so on.In this paper, SFC was applied to separation and purification of effective components from Chinese medicines and the mobile phase was SC-CO2 and a certain amount of modifier in the chromatographic separation process. And five Chinese medicines such as Angelica dahurica, Rheum officinale Baill., Aucklandia lappa Decne., and so on, were separated and purified by SFC. The purity of the obtained fractions was all above 97% determined by HPLC, and their chemical structures were identified by the nuclear magnetic resonance spectrum(NMR) and mass spectrometry(MS).1. Separation and purification of coumarins components from Angelica dahurica by SFC.In this study, the influence of separation effect of coumarins in angelica dahurica was investigated, which caused by three kinds of chromatographic column(YMC-C18 column, YMC-Diol column, YMC-NH2 column), four kinds of modifier(methanol, ethanol, isopropyl alcohol and acetonitrile), proportion of modifier, flow rate of mobile phase, pressure and temperature. The optimum chromatographic separation conditions were determined. And three coumarin compounds(imperatorin, oxypeucedanine, isoimperatorin) were obtained from 50 mg crude extract of Angelica dahurica, and their qualities respectively were 7.5 mg?10.2 mg and 5.8 mg. In addition, the thermodynamics of chromatographic process was studied. The result showed that under the experimental conditions, the SFC separation process was controlled by enthalpy.2. Separation and purification of active ingredients from Rheum officinale Baill. by SFC.A new method for separation and purification of five hydroxyanthraquinones and cinnamic acid from Rheum officinale Baill. by SFC was established. Through the optimization of experimental conditions, the optimal chromatographic separation conditions were obtained. Besides six compounds(I, II, III, IV, V, VI) were separated and purified from 50 mg crude extract of rhubarb and their qualities respectively were 4.9 mg, 8.9 mg, 7.8 mg, 4.1 mg, 4.1 mg and 2.7 mg.The thermodynamics of chromatographic process was also studied and it revealed that the chromatographic separation process was controlled by enthalpy. In addition, the retention behavior of the compounds on diol column in SFC separation process was discussed based on the chemical structures and the elution order. And the analysis showed that the retention of the compounds on diol column not only was related to the size of polarity, but also was related to strong or weak of hydrogen bonds interaction between the substance and diol groups. Therefore, it was a result of caused by the combining action of two kinds of factors.3. Separation and purification of the effective components from Aucklandia lappa Decne. by SFC.SFC was applied to separation of the active ingredients in Aucklandia lappa Decne. By optimizing the experiment conditions, the optimal SFC separation conditions were determined. And two kinds of sesquiterpene lactones(costunolide, dehydrocostus lactone) were separated and purified from from 80 mg crude extract of Aucklandia lappa Decne, and their qualities respectively were 34.7 mg and 25.3 mg.What's more, the thermodynamics of chromatographic process was also studied and it indicated that the chromatographic separation process was controlled by enthalpy. And it revealed that the separation process of SFC was normal phase chromatographic process under the experimental conditions, by combining the chemical structures and the elution order, compared with reversed phase high performance liquid chromatography(RP-HPLC).4. Separation and purification of the effective components from Ligusticum chuanxiong Hort. by SFC.In this study, SFC was applied to separation and purification of the lactones in Ligusticum chuanxiong Hort.. By optimizing the SFC separation conditions, the optimal conditions were determined. And two lactones(sedanolide, ligustilide) were separated and purified from from 60 mg crude extract of Ligusticum chuanxiong Hort., and their qualities respectively were 11.2 mg and 24.7 mg. Furthermore, the thermodynamics of chromatographic process was also studied and it indicated that the chromatographic separation process was controlled by enthalpy. And it revealed that the separation process of SFC was normal phase chromatographic process under the experimental conditions, based on the chemical structures and the elution order, compared with RP-HPLC.5. Separation and purification of active ingredients from Angelica pubescens Maxim.f. biserrata Shan ct Yuan by SFC.In this study, SFC was applied to separation and purification of the coumarins in Angelica pubescens Maxim.f. biserrata Shan ct Yuan. Through the optimization of experimental conditions, the optimal chromatographic separation conditions were obtained. And two coumarins(osthole, isoimperatorin) were separated and purified from from 50 mg crude extract of Angelica pubescens Maxim.f. biserrata Shan ct Yuan and their qualities respectively were 24.3 mg and 4.2 mg. Besides the thermodynamics of chromatographic process was also studied, and it indicated that the chromatographic separation process was controlled by enthalpy. And it revealed that the separation process of SFC was normal phase chromatographic process under the experimental conditions, based on the chemical structures and the elution order, compared with RP-HPLC. |
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