【作者】 陈华山；

【导师】 李发美；

【作者基本信息】 沈阳药科大学 ， 药物分析学， 2006， 硕士

【摘要】 天山雪莲收载于《中国药典》（2005年版一部），其药理作用有抗炎镇痛、抗衰老及抑制癌细胞增生等。随着天山雪莲现代药理研究的深入及其在抗风湿、类风湿性关节炎等临床上的应用，天山雪莲日益引起国内外药学工作者的关注。临床上已有雪莲注射液用于风湿性关节炎、类风湿性关节炎，疗效确切、可靠、毒副作用小。以天山雪莲药材为原料，采用现代科学技术手段，制成冻干粉针能够增强其的稳定性，因此研制了注射用雪莲（冻干）。本论文研究内容包括：（1）注射用雪莲（冻干）的制备；（2）注射用雪莲（冻干）指纹图谱的研究；（3）注射用雪莲（冻干）中指标成分分析方法及质量标准的建立；（4）注射用雪莲（冻干）稳定性研究；（5）注射用雪莲（冻干）在大鼠体内的药物动力学研究。经处方筛选和工艺优化，以天山雪莲提取液1000mL为原料，以150g甘露醇为支持剂，-40℃预冻5小时，-20℃减压干燥（＜10Pa）5小时，将温度升至20℃，继续减压干燥6小时制得产品外形饱满、疏松，成型性好。使用C₁₈色谱柱，以乙腈-50mmol·L^-1KH₂PO₄水溶液（H₃PO₄调pH3.6）为流动相进行梯度洗脱，紫外法检测，建立了体现注射用雪莲（冻干）化学成分特征的HPLC指纹图谱。选用紫丁香苷为参照物，通过对10批注射用雪莲（冻干）供试品指纹图谱的分析，确定了13个共有指纹峰，建立了共有模式。相似度分析表明，10批供试品的相似度均大于0.95，说明注射用雪莲（冻干）制备工艺稳定，批间重现好。制剂、中间体和药材的指纹图谱具有很好的相关性。采用简便的TLC鉴别方法，以对照药材和芦丁为指标，对制剂进行了鉴别。采用高效液相色谱法和比色法建立了芦丁和总黄酮两种指标性成分的含量测定方法，方法可靠，重现性好。加速试验和留样试验结果表明，制剂的稳定性良好。建立了测定大鼠血浆中芦丁含量的反相高效液相色谱分析方法，以新北美圣草苷为内标，血浆经甲醇沉淀蛋白后进样分析。芦丁在0.050～3.2μg·mL^-1（r=0.9901）范围内线性关系良好。方法的定量下限为0.050μg·mL^-1。测定了芦丁的血药浓度-时间曲线，计算其相应的药物动力学参数。注射用雪莲（冻干）腹腔给药后，芦丁的C_max为1.410μg·mL^-1，t_max为8.0 min，t_1／2为84.2 min，AUC_0-∞为131.6 mg·L^-1·min。更多还原

【Abstract】 Saussurea involucrate （ Kar.et Kir.） Sch. Bip., listed in《Phamacopoeia of the People’s Republic of China》（Edition 2005,Volumn I ） is a kind of ethnic medicine, possessing anti-inflamamation, analgesia, anti-aging and contraception. With its clinical application to the treatment rheumatic arthritis and rheumatoid arthritis, Saussurea involucrate （ Kar.et Kir.） Sch. Bip. increasingly attracts the attention of pharmaceutical researchers. Xuelian injection has definite therapeutic effectiveness. If Xuelian is prepared into freeze dried powder with modern technology, its stability must be enhanced. This study includes: （1） the preparation design of Xuelian for injection （2） the HPLC-finger-printing development of Xuelian for injection （3） quality control method of Xuelian for injection （4） the stability test of Xuelian for injection （5） the pharmacokinetics of Xuelian for injection in rats.The prescription and technological process were optimized by the criterion of appearance, moisture and resolubility. An good product was obtained with 300 mg mannitol each bottle. The optimized technological process is as follows: pre-freeze for 5 hours at -40℃, and then for 6 hours under vacuum at -20℃, raising the temperature to 20℃and drying for another 6 hours under vacuum.An HPLC-FP of Xuelian for injection was developed with C₁₈ column employingCH₃CN-50mmol·L^-1KH₂PO₄（pH=3.6） as mobile phase for gradient elution with ultraviolet detection. Thirteen common peaks were achieved after analyzing 10 batches of Xuelian for injection, Syringin was choosed as the reference substance, and the HPLC-FP common pattern of Xuelian for injection was developed. The similarities of 10 batches of the samples were all above 0.95, indicating the preparation process of Xuelian for injection is reproducible. There is a good correlationship among HPLC-FP of Xuelian for injection, intermediate and Saussurea involucrate （ Kar.et Kir.） Sch. Bip..Standard herbs and rutin, as typical principles were qualitalively analyzed with simple and proper TLC methods. An HPLC method was developed to determine the content of rutin in the preparation. Flavonoids in the preparation were determined colourmetrically. All these methods are accurate and sensitive.The preparation was stable in the accelerated test and at room temperature.An RP-HPLC method was developed for the determination of rutin in plasma of rats after treated with Xuelian for injection intraperitoneaUy. After added Neoeriocitrin as internal standard, plasma samples were extracted with methanol. The linear range for rutin was 0.050～3.2μg·mL^-1 （r=0.9901）. The LLOQ was 0.050μg·mL^-1. Both accuracy and precision of the assay were satisfactory. The plasma concentration-time curve of rutin was plotted and the pharmacokinetic parameters were calculated.The C_max, t_max, t_1/2 and AUC_0-∞ of rutin in rats were 1.410μg·mL^-1, 8.0 min, 84.2 min and 131.6 mg·L^-1·min, respectively.更多还原