【作者】 周国勤；

【导师】 王淑君；

【作者基本信息】 沈阳药科大学 ， 药物制剂， 2009， 硕士

【摘要】 藏药独一味(Lamiophlomis rotate (Benth.)Kudo)为藏族习用药材,具有活血止血,祛风止痛,干黄水等功效。近年来,独一味由于其疗效好,迄今未发现毒副作用而广泛应用。文献报道独一味总黄酮是独一味镇痛的有效成分,临床可用于治疗骨科骨折引起的疼痛,风湿痛等。有报道研究表明藏药独一味作为癌痛第一阶梯的止痛药,可避免由使用吲哚美辛产生的副作用。首先建立了独一味总黄酮和黄酮类成分木犀草素的检测方法,为后来的提取分离和制剂工作提供依据。建立紫外.可见分光光度法测定总黄酮的含量,采用亚硝酸钠、硝酸铝、氢氧化钠显色法以芦丁为对照品计算总黄酮的含量,标准曲线方程为A=12.07C+0.0057(r=0.9998),芦丁浓度在0.019～0.069 mg.mL-1之间浓度与吸光度具有良好的线性关系。精密度实验、稳定性试验、回收率实验等结果显示此方法可以准确、可靠地测定独一味总黄酮的含量。同时建立高效液相色谱法测定制剂中木犀草素的含量,色谱条件为：色谱柱：迪马Diamonsil(TM)钻石柱(C18,5μm,4.6mm×200mm)；流动相：甲醇-乙睛-0.1%甲酸溶液(4：1：5)；流速：1.0 mL·min-1;检测波长：350 nm；柱温：室温；进样量：5μL。标准曲线方程为A=33786C+6099(r=0.9999),木犀草素浓度在4.26～21.28μg·mL-1范围内进样量与峰面积之间具有良好的线性关系。精密度实验、稳定性试验、回收率实验等结果显示,此方法可以准确、可靠地测定制剂中木犀草素的含量。通过单因素实验和正交实验优化独一味总黄酮的制备工艺。通过提取方法、提取溶剂、粉碎粒度、浸泡时间等单因素考察和提取溶剂用量、提取次数、提取时间的三因素三水平正交实验优化提取工艺,最佳提取工艺为：药材粉碎,过40目筛,以pH值为9的氢氧化钙溶液配制的50%乙醇溶液为提取溶剂,浸泡12 h,10倍量溶剂回流提取3次,提取时间为1 h。通过石油醚萃取、上样浓度考察、上样速率考察、洗脱溶剂考察、洗脱体积考察等结果确定最佳分离富集工艺。最佳分离富集工艺为：提取液经石油醚萃取后配制成15mg·mL-1的上柱液,以1 BV·h-1的速度上样,预吸附2小时,过柱液重吸附一次,静置吸附24 h。用大量蒸馏水洗脱至洗脱液无色,再以50%乙醇溶液洗脱4 BV,收集50%乙醇洗脱液,浓缩,干燥,即得独一味总黄酮精制中间体。通过药理实验验证独一味总黄酮精制中间体的镇痛和止血作用。通过醋酸扭体实验和热板实验考察了水提、醇提、纯化不同制备方法所得物的镇痛效果,结果显示镇痛效果与总黄酮的量具有潜在的量效关系。通过毛细管法和断尾止血实验,结果显示独一味精制中间体具有一定的凝血作用。充液胶囊以全新概念的固体剂型液体释放而备受关注,被称为“液体和半固体制剂的理想容器”。各种液体或半固体状天然植物提取物,均可采用该技术。本文通过稀释剂的选择,稀释剂PEG400含水量的考察,助悬剂的选择和用量的考察,封口乙醇浓度的考察等实验优化独一味充液胶囊处方和工艺。其处方为：稀释剂为含水量10%的PEG400,稀释剂、助悬剂(HPMC-K15M)和药物的比例为48：2：6。抑菌剂为0.15%的尼泊金乙酯,用50%的乙醇溶液封口。溶出度实验结果显示独一味充液胶囊在20 min内的累积溶出度可达80%以上。稳定性试验结果显示,制剂在常温下放置稳定。滴丸剂为中药提取物的一种良好的剂形,药物高度分散在基质中,可以提高药物的溶出度和生物利用度。通过基质选择、载药量的考察、冷凝剂的选择等单因素实验和药液温度、滴距、冷凝剂高度三因素三水平正交实验筛选最佳处方和工艺。最佳处方和工艺为：PEG4000和PEG6000联合应用为基质、比例为2：1；载药量为30%；药物与基质85℃熔融混合并保温；冷凝剂为20：1的液体石蜡与蜂蜡混合物,冷凝剂高度为40 cm,下端15 cm以0-5℃的冰水浴冷却；滴口内径2.5 mm；滴距为5 cm；滴速20 d·min-1。溶出度实验结果显示独一味滴丸在20 min内的累积溶出度可达85%以上。稳定性试验结果显示,制剂在常温下放置稳定。更多还原

【Abstract】 Lamioplomis rotata (Benth.) Kudo widely grows in alpine meadow and wet land at altitude 4000 m. It distributes in Tibet, Qinghai, Yunnan, Sichuan, Gansu province, commonly used as a medicinal herb by Tibet, Mongolia and Naxi people to promote blood circulation, remove blood stasis, subdue swelling and alleviate pain. That main compounds in lamiophlomis were reported as flavonoids, iridoids and phenethanol glycoside. It is reported that total flavonoids of lamiophlomis is the chief component to alleviate pain, which can be used to cure pain caused by bone fracture, rheumatalgia, etc. Because of its good effect and little side effect, it is largely used. Some report indicates that it can be used to alleviate pain at the first stage of cancer, which can avoid side effect caused by indomethacin.we have respectively researched the effective components in Lamiophlomis, then established methods of determining the total flavonoids and luteolin, as well as the preparing technology of Du Yiwei liquid hard capsules and Du Yiwei dripping pills.The influences of different factors and levels on the preparation process were investigated by means of single factor test and orthogonal test. We chose transfer rate and purities of total flavonoids as the assessment index. Through the study of preparation process, we established the optimum conditions for preparation process, including such steps as using the 50% ethanol mixed with Calcium Hydroxide solution (pH=9) as solvent(the ratio of liquid to solid material is 10:1); respectively hot reflux extraction 4 times, once for 1 hour; the solution concentrating at 15 mg·mL-1, separating components by macroporous adsorptive resin; at a adsorption rate of 1 BV·h-1, using 50% ethanol as the eluting reagent, with eluent volume of 4 BV. drying through vaporation to get intermediates, with purity of total flavonoids at 58.64%.UV spectrophotometry was established to determine the total flavonoids, the content of the total flavonoids was calculated with rutin as standard substance; the HPLC was established to determine the luteolin.The effect of alleviating pain was confirmed by acetic acid writhing and hot plate test in mice, from the test we know that the total flavonoids has significant analgesic action, and the effect of alleviating pain was dependent on the dose of total flavonoids.To optimize the formulation of Du Yiwei liquid hard capsules. A series tests were carried out for the selection of diluent agent and suspending agent. The balanced amount of water and the best proportion was obtained with PEG400 (amount of water was 10%) as diluent agent. The lamiophlomis liquid hard capsules were prepared with the mixture of diluent agent, HPMC-K15M and drug (the ratio is 48:2:6), which was dissolved quickly and adequately.We studied the formula and forming technology of Du Ywei dripping pills according to the results of the preliminary test。The single factor test and orthogonal test was carried out to optimize the formula with the degree of circularity and hardness as index. Then we established the best formula and preparing technique as following:the dispersing agent was PEG4000/PEG6000 (3:1), the ratio of drug and dispersing agent was 1:2; the melting temperature was 95℃; the condensate is liquid paraffin, the temperature of refrigerant was 0～5℃. Through the stability test, the result showed that the Du Yiwei dripping pills was stable, the quality was controllable.更多还原