【作者】 黄敬群；

【导师】 黄熙；

【作者基本信息】 中国人民解放军第四军医大学 ， 中西医结合临床， 2003， 硕士

【副题名】丹参复方的制剂学监测及冠心Ⅱ号来源的阿魏酸血清浓度测定的方法学

【摘要】 从二十世纪六十年代起，“治疗药物监测”广泛用于监测病人体液中的药物浓度，并研究浓度与效应关系，以利于制订个体化用药方案，减轻毒副作用，达到最佳疗效。无论如何，上面提到的思路或方法很少或没有用在中医临床。在对“证治药动学”长期研究的基础上，黄熙教授提出了“方剂治疗药物监测”的设想。旨在通过对方剂制剂学、药动学、药效学、治疗学的全面监测，建立可靠的方剂质量控制标准，阐明方剂药效物质、方剂配伍理论的科学内涵，尽可能地实施个体化、量化的辨证施治方案，以提高临床疗效和减轻毒副作用。 本文首先进行了丹参复方的制剂学监测(监测吸收入体内的成分)及冠心Ⅱ号来源的阿魏酸血清浓度测定的方法学研究。上述研究结果有助于方剂治疗药物监测的进一步研究。 目的： 为了初步探索方剂治疗药物监测中的制剂学和药动学中的方法学，本实验选择植物化学、药理与临床研究较为成熟的丹参及其复方冠心＊号为工具，在对丹参复方制剂学监测的基础上，建立RP－HPLC测定人血清中阿魏酸浓度的改进方法。方法与结果：1．高效液相色谱法测定不同煎煮条件下丹参和冠心＊号中丹参素、原儿茶醛的含量： （l）方法：反相高效液相色谱法。Kromasil 分析柱p50nun X 46rum，sum，No：C18B020307），流动相：乙胎一0刀…帅·L’磷酸二氢钾溶液汐：gi），磷酸调至pH 2．8，紫外检测波长：280urn，流速：l＊rnL·Inn1。 （2）结果：丹参素在 0．245～5．880哈（F09998），原儿茶醛在 0＊＊ ～0．504ty（r＝0．9996）范围内呈线性，丹参素和原儿茶醛最低检测浓度分别为9．SM”血－’和2．IM·血－’，回收率分别为93．24％（RSD 为2．53％）和102＊8％（RSD为1．gi％）。 门）结论：中药的煎煮时间和加水量对中药有效成分的煎出量均有影响。在煎煮时间相同的情况下，无论是丹参素还是原儿茶醛，加 12倍水的溶出率都高于加8倍水；随着煎煮时间的延长，丹参素和原儿茶醛的溶出率逐渐增高；冠心＊号煎剂五味药合煎，在任何加水量，任何煎煮时间条件下，其丹参素和原儿茶醛的溶出率都明显高于相同条件下的丹参煎剂。2．高效液相色谱法测定五种不同产地丹参中丹参素和原儿茶醛含量： （二）方法：反相高效液相色谱法。Ko——】分析柱仅50mx 46llllll－slirD．NO汇18B 02lllo），流动相：甲醇一1％冰醋酸 20：80，紫外检测波长：280urn，流速：1．oml．·a‘。 u）结果：丹参素在4．2－33．6ng·mL‘，原儿茶醛在0．36～l．SN·InL－‘范围内线性关系良好，丹参素和原儿茶醛最低检测限分别为10．sng和0．6ng。回收率分别为 99．36％（RSD为 l．289％）和 97．56％（RSD为 0．774＄）。 门）结论：丹参素的含量（陀·mL’）卢氏野生丹参＞洛阳丹参冲氏家种丹参＞蒲城丹参＞陕南丹参。原儿茶醛的含量（卜g·mL－’）蒲城丹参＞洛阳丹参＞卢氏野生丹参＞卢氏家种丹参＞陕南丹参。3．RI，nPLC测定人血清中阿魏酸浓度的改进方法： （）方法：y丑PLC法，采用 lro…il（ 柱为分析柱，甲醇－水－冰醋酸 （3．4：63：0石）为流动相，流速1刀InL·min－’，检测波长sHnm，内标物为对羟基苯甲醛。用乙脂既去除人血清中的蛋白，又提取血清中的阿魏酸。 Q）结果：阿魏酸在 9．94－159刀4ng伽L’范围内与峰面积呈良好线性关系 （r＝0．9925），日内和日间变异系数均＜10％，平均回收率为99．77％，最低检测浓度为sllg·InL‘。 （3）结论：该法简便、快速、灵敏、重现性好，低毒，适用于人血清中阿魏酸的定量分析。更多还原

【Abstract】 Since 1960’s, Therapeutic drug monitoring (TDM) has been widely used to monitor the body fluid’s drug concentration of patients. TDM studied the relationship between the concentration and effect after the administration of drug to men and adjusted individual dosage regimen so as to get the best therapeutic effectiveness as well as decrease the toxicity and side-effect of that drug [1] However, TDM has little or never used to the clinical treatment of Traditional Chinese Medicine (TCM) so far. On the basis of study on the relationship between TCM Syndrome (terminology on traditional Chinese medicine) and pharmacokinetics(PK) for a long time[2,3], Professor Huang Xi put forward the idea about Recipe TDM [4] recently. Recipe means a terminology about pharmacology of traditional Chinese medical formulae. Recipe TDM consist of Pharmaceutics (PKT), pharmacokinetics (PK), pharmacodynamics and therapeutics monitoring. By monitoring these contents, it helps to develop the standard of quality controlling of Traditional Chinese Recipe(TCR) and elucidate the scientific mechanism of TCR. It is also adjusted individual dosage regimen so as to increase the therapeutic effectiveness as well as decrease the toxicity andside-effect of that recipe.This experiment primely monitor the pharmaceutics of Salvia Miltiorrhiza Bge prescription and develop the improved methodology of detecting ferulic acid serum concentration after oral administration of Coronary Heart No II Decoction to healthy human. Above results help to further study recipe TDM. ObjectiveTo initially explore the PKT and PK methodology of recipe TDM , This work select the Salvia Miltiorrhiza Bge and one of it’s prescription Coronary Heart No II dcoction as the research tool, Which accumulated many evidences in phytochemistry, pharmacology and clinical medicine. On the basis of PKT monitoring of Salvia Miltiorrhiza Bge prescription, develop the improved RP-HPLC method for the determination of Ferulic Acid concentration in human serum.Methods and results1. Danshensu and Protocatechuic Aidehyde contents of Salvia Miltiorrhiza Bge and Coronary Heart No II Decoction in Different Decocting Methods are Quantitatively determined by HPLC respectively.(1) MethodsrChromatographic conditions including Kromasil column (250mm X 4 6mm,5nm), the mobile phase consists of acetonitrile and 0.01 moL ·L-1 potassium dihydrogen phosphate (9:91,adjusted to pH 2.8 with phosphoric acid), the UV detector was set at 280 nm and the flow rate was 1.0 mL·min-1.(2) Results:The result showed that the standard curve of Danshensu was linear over the range of 0.245~5.880ug(r=0.9998); Protocatechuic Aidehyde was linear over the range of 0.011 ~0.504μg(r=0.9996).(3) Conclusion: Both amount of water added and decocting duration required had an influence on the output of the concents of TCM by decocting. At the same decocting duration required, When added 12 times water, the output of the Danshensu or Protocatechuic Aidehyde by decocting are higher than that of added 8 times water. The decocting duration required longer, the output of Danshensu or Protocatechuic Aidehyde by decocting higher. When the five crude drugs of Coronary Heart No II decoction boiling together, whichever amount of water added or decocting duration required, the output of the Danshensu or Protocatechuic Aidehyde by decocting are awalys higher than that of Salvia Miltiorrhiza Bge’s dcoction.2. Danshensu and Protocatechuic Aidehyde contents of Salvia Miltiorrhiza Bge in five place of production are Quantitatively determined by HPLC respectively.(1) MethodsrChromatographic conditions including Kromasil column (250mm X 4 6mm,5nm), the mobile phase consists of methanol and 1% Acetic acid (20:80), the UV detector was set at 280 run and the flow rate was 1.0 mL· min-1.(2) Results: The result showed that the standard curve of Danshensu was linear over the range of 4.2~33.6#g·mL-1, Protocate Chualdehyde was linear over the range of 0.36~1.8#g·mL-1’.(3) Conclusion: The quant更多还原