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1、超临界流体色谱法应用于几种药物有效成分分析的研究昆明医学院药学院 研究生：杨 敏 导 师：郭亚东 教授中文摘要目的：探讨超临界流体色谱法（Supercritical Fluid Chromatography，SFC）应用于两种西药（阿咖酚散和去痛片）以及三种中药（厚朴、吴茱萸和木香）有效成分分析的研究，并对其含量进行测定。研究内容：1.超临界流体色谱法分离测定阿咖酚散中阿司匹林、咖啡因和对乙酰氨基酚的研究；2.超临界流体色谱法分离测定去痛片中苯巴比妥、非那西丁、咖啡因和氨基比林的研究；3.超临界流体色谱法分离测定厚朴中厚朴酚和和厚朴酚的研究；4.超临界流体色谱法分离测定吴茱萸中吴茱萸次碱和吴茱
2、萸碱的研究；5.超临界流体色谱法分离测定木香中去氢木香内酯和木香内酯的研究。 实验方法：采用单因素试验法，以厚朴、吴茱萸和木香中主要有效成分的含量为评价指标，考察超声提取法和微波提取法中温度、时间、料液比对提取效果的影响，对提取方法的总体效果进行比较。使用超临界流体色谱仪，以加入适量甲醇作改性剂的超临界CO2为流动相，考察流动相组成、温度、压力等对待侧组分分离的影响，并以优化的色谱条件测定阿咖酚散、去痛片以及厚朴、吴茱萸、木香中有效成分的含量。结果：通过超临界流体色谱分析条件摸索，阿咖酚散使用Kromasil Slica填充柱（250 4.6 mm, 5 m），流动相为含18 %甲醇的CO2，
3、流速2.0 mLmin-1，柱温50 ，背压200 bar，检测波长275 nm；去痛片使用Kromasil Slica填充柱（250 4.6 mm, 5 m），流动相为含21 %甲醇的CO2，流速1.0 mLmin-1，柱温60 ，背压150 bar，检测波长214 nm；厚朴使用Kromasil Slica填充柱（250 4.6 mm, 5 m），流动相为含5 %甲醇的CO2，流速1.0 mLmin-1，柱温60 ，背压200 bar，检测波长294 nm；吴茱萸使用Kromasil Slica填充柱（250 4.6 mm, 5 m），流动相为含13 %甲醇的CO2，流速2.0 mLmin
4、-1，柱温60 ，背压200 bar，检测波长225 nm；木香使用LiChrospher 100 Amino填充柱（250 4.6 mm, 5 m），流动相为含4 %甲醇的CO2，流速1.0 mLmin-1，柱温60 ，背压130 bar，检测波长225 nm。 结论：本研究首次采用超临界流体色谱法分离测定阿咖酚散、去痛片以及厚朴、吴茱萸、木香中的主要有效成分，方法简便快速、条件温和。同时，本研究所建立的含量测定方法，具有分离效果佳、系统适应性好、结果稳定等特点，可为上述五种药物（药材）的质量控制方法提供参考。 关键词：超临界流体色谱法 阿咖酚散 去痛片 厚朴 吴茱萸 木香Study on
5、application of supercritical fluid chromatography in 5 drugsFaculty of Pharmaceutical science，Kunming Medical CollegePostgraduate：Yang MinAcedemic supervisor：prof.Guo YadongAbstractObjective：To investigate the application of supercritical fluid chromatography in 2 chemicals and 3 chinese herbal drug
6、s，and to determinate the contents of effective components in these 5 drugs by SFC.Contents：1.To establish a SFC method to determinate the three components in Akafen powder. 2.To establish a SFC method to determinate the four components in Qutong tablet. 3.To establish a SFC method to determinate the
7、 magnolol and honokiol in Officinal Magnolia Bark. 4.To establish a SFC method to determinate the rutaecarpine and evodiamine in Medicinal Evodia Immature Frui. 5.To establish a SFC method to determinate the dehydrocostuslactone and costunolide in Costusroot.Method：By taking the extraction rates of
8、the main effective components in the 3 chinese herbal drugs as the evaluation index，single factor experiments were adopted to observe the influences on ultrasound extraction and microwave extraction，temperature，extracting time and methanol volume were selected as the examing factors. To study the ef
9、fects of flow rate, modifier concentration, back pressure and column oven temperature on chromatographic resolution，to determinate the contents of effective components in these 5 drugs by optimal SFC method. Result：The optimal SFC parameters were using the Kromasil Slica column (250 4.6 mm i.d. , 5
10、m) and setting back pressure of 200 bar, column oven temperature of 50 C，Modifier concentration of 18 % and flow rate of 2.0 mLmin-1 for Akafen powder. Using the Kromasil Slica column (250 4.6 mm i.d. , 5 m) and setting back pressure of 150 bar, column oven temperature of 60 C，Modifier concentration
11、 of 21 % and flow rate of 1.0 mLmin-1 for Qutong tablet. Using the Kromasil Slica column (250 4.6 mm i.d. , 5 m) and setting back pressure of 200 bar, column oven temperature of 60 C，Modifier concentration of 5 % and flow rate of 1.0 mLmin-1 for Officinal Magnolia Bark. Using the Kromasil Slica colu
12、mn (250 4.6 mm i.d. , 5 m) and setting back pressure of 200 bar, column oven temperature of 60 C，Modifier concentration of 13 % and flow rate of 2.0 mLmin-1 for Medicinal Evodia Immature Frui. Using the LiChrospher 100 Amino column (250 4.6 mm i.d. , 5 m) and setting back pressure of 130 bar, column
13、 oven temperature of 60 C，Modifier concentration of 4 % and flow rate of 1.0 mLmin-1 for Costusroot. Conclusion：The packed column SFC method have the advantage of quick analysis time, simple preparation of sample, low-cost organic solvent consumption and environment-friendly, etc. Besides, using sup
14、ercritical carbon dioxide as mobile phase modified with little organic solvent help extend the useful life of packed column. In a word, The introduction of SFC to the area of traditional Chinese medicine is very promising.Key words：Supercritical Fluid Chromatography; Akafen powder; Qutong tablet; Officinal Magnolia Bark; Medicinal Evodia Immature Frui; Costusroot.
