【作者】 谢昀；

【导师】 于治国；

【作者基本信息】 沈阳药科大学 ， 药物分析， 2005， 硕士

【摘要】 黄杨木作为药用，在《本草纲目》中已有记载，民间用其治疗心血管病。黄杨木中主要活性成分环维黄杨星D（商品名为黄杨宁），目前已从黄杨木中提取分离，并与其制剂黄杨宁片一起收载于中国药典（2000年版）。本文建立了环维黄杨星D的反相高效液相色谱测定法；测定了不同产地的黄杨木药材中环维黄杨星D的含量并建立了黄杨木药材的质量标准；对黄杨木中提取分离环维黄杨星D的方法进行了改进；研制开发了环维黄杨星D分散片并制定了质量标准。1．环维黄杨星D含量测定方法的研究中国药典中对环维黄杨星D原料及片剂的含量测定，分别采用非水滴定法和分光光度法；文献报道的方法有HPLC／MS法、荧光或紫外衍生化HPLC法。本文建立了一种测定环维黄杨星D的液相方法，简便快速，不需衍生化，方法专属性和重现性好。该方法使用ODS柱，流动相为0.005mol·L^-1庚烷磺酸钠溶液（内含0.15％的三乙胺，磷酸调节pH至3.1）-乙腈（73：27，ν／ν），检测波长203nm。环维黄杨星D在0.5～8.0μg范围内峰面积线性关系良好（r=0.9999），方法重现性RSD为0.9％，平均回收率为98.6％。测定了3批不同产地的市售环维黄杨星D原料，结果含量约为85％。2．黄杨木中环维黄杨星D的含量测定及质量标准研究建立了黄杨木中环维黄杨星D的含量测定方法。样品用氯仿回流提取，经氯仿和乙酸乙酯预处理后，在ODS柱上分离，流动相为0.005mol·L^-1十二烷基硫酸钠溶液（内含0.15％的三乙胺，磷酸调节pH至3.1）-乙腈（54：46，ν／ν），检测波长为203nm，柱温45℃。在1.24～12.4μg范围内，峰面积线性关系良好（r=0.9993），方法平均加样回收率为98.1％，重现性RSD为1.5％。本法结果准确、重现性良好，适用于黄杨木中环维黄杨星D的含量测定。在此基础上，建立了黄杨木药材的质量标准。采用TLC法鉴别黄杨木中的环维黄杨星D：使用硅胶G板，以乙酸乙酯-甲醇-氨水（17：2：1，ν／ν／ν）为展开剂，改良碘化铋钾为显色剂。采用上述HPLC法测定黄杨木中环维黄杨星D的含量，要求不得少于0.090％（g／g）。3．环维黄杨星D制备工艺的改进采用环己烷溶剂提取法，通过正交试验，优化提取分离条件，确定以氨水与95％乙醇的混合液（pH=10）为碱化溶液，碱化1h后提取。以磷酸二氢钠缓冲液（pH4.0）为沉淀剂，制得粗品；粗品使用氯仿-丙酮重结晶得最终产品。经HPLC法测定，产品纯度为89％。4．环维黄杨星D分散片的制备和质量标准研究通过处方筛选，确定以交联聚维酮为崩解剂，乳糖和微晶纤维素为填充剂，选择亲水性粘合剂PVPK₃₀；崩解剂的加入采用内外加法。制成的分散片在30s内完全崩解并通过2号筛，在5分钟时溶出度达到95％以上。制订了环维黄杨星D分散片的质量标准，选择与环维黄杨星D含量测定相同的HPLC法，测定分散片中环维黄杨星D的含量。建立了一种灵敏度较高的HPLC-ELSD法测定溶出度，色谱条件：ODS柱，流动相为0.2％的甲酸和0.2％的三乙胺溶液-乙睛（80∶20，v／v），流速0.8mL·min^-1；检测器条件：漂移管温度105℃，气体流速2.8 L·min^-1。更多还原

【Abstract】 1. Studies On the determination methods of cyclovirobuxium DIn this paper a method using IP-HPLC was developed to determine cyclovirobuxium D . The assay was conducted on a C₁₈ column with acetonitrile -0.005 mol·L^-1 solution of 1-Heptanesulfonic acid sodium （mixing 0.15% TEA and modify pH to 3.1 with H₃PO₄） （27:73, v/v） as mobile phase. The detection wavelength was 203 nm. Cyclovirobuxium D obtained good isolation from its related substances. The calibration curve was linear in the range of 0.5⁸.0μg of cyclovirobuxium D with correlation coefficient 0.9999. The average recovery of cyclovirobuxium D was 98.6%, RSD was 0.9%.Three batches of cyclovirobuxium D from different source were determined and the determination results was about 85%.2. The determination of cyclovirobuxium D in huangyangmu and quality specification of huangyangmuA rapid and accurate liquid chromatography method was establiahed to determine cyclovirobuxium D in huangyangmu. Using chloroform as extracting solvent, the sample of huangyangmu was refluxed for 3 hours. Then the extract was pretreated by extraction separation ,with chloroform and ethyl acetate as extractant. The assay was carried out on C₁₈ column, the mobile phase was acetonitrile-0.005mol·L^-1 solution of sodium dodecylsulfate （mixing 0.15% TEA and modify pH to 3.1 with H₃PO₄） （46:54, v/v）. The detection wavelength was set at 203 nm, column temperature was 45℃. The calibration curve was linear in the range of 1.24-12.4μg of cyclovirobuxium D with correlation coefficient 0.9993. The average recovery of cyclovirobuxium D was 98.1%, RSD was 1.5%. The method was reliable and could be used for the determination of cyclovirobuxium D in huangyangmu with satisfactory reproducibility.On this base, the quality specification of huangyangmu was worked out. The cyclovirobuxium D in huanyangmu was identified by TLC with modified development system, using silica gel G plate, ethyl acetate-methanol-ammonia water （17:2:1,v/v/v） as developing solvent. The content of cyclovirobuxium D in huangyangmu should not be less than 0.090%（g/g） using the above liquid chromatography method.3. The preparation process of cyclovirobuxium DUsing cyclohexane as the extraction solvent, the optimal extraction methods were studied by orthogonal design with yield of raw crystal as index. The result showed that the optimal alkalization condition was the mixing solution of 95% alcohol and ammonia water（pH=10） as alkalization solvent, alkacified for 1 hour. Then the sodium dihydrogen phosphate buffer（pH4.0） was used as precipitant to obtain more depurated crystal; the product of cyclovirobuxium D was obtained through recrystallization process with chloroform-acetone as recrystallization solvent. The purity of cyclovirobuxium D product was 89%.4. The preparation process and quality specification of cyelovirobuxium D dispersible tabletsCyclovirobuxium D dispersible tablets was prepared with crospovidone as disintegrant, lactose and microcrystaUline cellulose as fillers. A sensitive HPLC-ELSD method was developed to determine the dissolution of cyclovirobuxium D in the tablets. Chromatographic condition included mobile phase water（mixing 0.2% formic acid and 0.2% TEA）: acetonitrile（80: 20, v/v）, flow rate 0.8 mL·min^-1, drift tube temperature 105℃, gas flow 2.8 L·min^-1. The result showed that the dissolution of cyclovirobuxium D was more than 95%. The quality specification of cyclovirobuxium D dispersible tablets was also established.更多还原