摘要
本文选用万氏牛黄清心丸作为模型药物，研究了方中诸药有效部位的提取工艺，建立了检测内在质量的定性定量方法，在此基础上研究制备了万氏牛黄清心缓释微丸。
采用TLC方法，对万氏牛黄清心缓释微丸中有效成分盐酸小檗碱、黄芩苷、栀子苷进行了定性鉴别；采用HPLC法对样品中盐酸小檗碱、黄芩苷、栀子苷进行了含量测定，并测定了肠液中盐酸小檗碱和酚红的浓度，建立了测定家兔血浆中盐酸小檗碱浓度的方法。
通过单因素考察实验和正交实验设计法分别确定了黄连、黄芩、栀子、郁金的最佳提取工艺；采用β-CD对提取的挥发油进行了包合研究，并通过均匀实验设计法进行了工艺的优化。
对壳聚糖的脱乙酰度和分子量进行了测定；以壳聚糖为原料制备了羟丙基壳聚糖。通过单因素实验，考察了碱化、反应温度、反应时间、反应介质对产物取代度的影响，并对产物进行了定性鉴别，考察了产物的水溶性和特性粘度。
采用大鼠在体灌流方案，通过不同肠段的结扎对盐酸小檗碱的吸收部位和吸收动力学进行实验研究。实验结果表明：盐酸小檗碱在肠道的吸收较差，主要集中在小肠的上部吸收，按十二指肠、空肠、回肠和结肠的顺序，吸收百分率和吸收速率常数K_a依次降低。各肠段间吸收百分率和吸收速率常数有显著差异。药物在肠道内的吸收机制是被动扩散的方式。
以MCC为稀释剂，3％HPMC水溶液为粘合剂，采用离心造粒法，在优化条件下可制得表面较为光滑、圆整度较高的万氏牛黄清心微丸。体外
沈阳药科大学硕士学位论文 摘要
溶出实验表明，微丸中盐酸小梁碱的溶出度符合制剂要求。
以自制的羟丙基壳聚糖和pH依赖型辅料丙烯酸树脂的水分散体作为
包衣材料制备了3种包衣微丸，体外模拟人体胃肠道pH条件下测定盐酸小
巢碱的释放度，结果表明释药呈现出一种pH依赖型梯度释药特征。
家兔口服自制的万氏牛黄清心缓释胶囊后盐酸小凳碱的血药浓度测定
结果表明，盐酸小梁碱的峰值血药浓度Cm。明显降低，作用时间显著延长，
体内也呈现了预期的梯度脉冲释药特征。
Wanshi Niuhuang Qingxin pills(WSNHQXP) was selected as model drug and the preparation techniques of active fraction of medicinal materials in prescription were investigated. The qualitative and quantitative analysis methods were developed to control the inherent quality of formulation. Based on these results, Wanshi Niuhuang Qingxin sustained-release pellets(WSNHQX SRP) were investigated.
TLC methods were used to identify Berberine Hydrochloride, Baicalin, Geniposide in WSNHQX SRP; the method of HPLC for Berberine Hydrochloride, Baicalin, Geniposide were developed to determine their contents in preparation. The concentration of Berberine Hydrochloride and phenolsulfonphthalein in intestine solution and concentration of Berberine Hydrochloride in rabbit plasma were also assayed by HPLC method.
The extraction techniques of Coptis chinensis Franch, Scutellaria baicalensis Georgi, Gardenia jasminoides Ellis, Curcuma wenyujinY.H.Chen et C.Ling were optimized by orthogonal experiment design method and single-factor test method, respectively. The b-CD inclusion complex of Zedoary Turmeric oil was formulated by the method of uniform experiment design.
Deacetylation degree and molecular weight of chitosan were determined. Chitosan was selected as a material to synthesize hydropropylated chitosan and the effects of alkalization, reaction temperature, time, solvent on degrees of
substitution were studied. Hydropropylated chitosan was identified, solubility and intrinsic viscosity were investigated, too.
The absorption sites and the absorption kinetics were studied from various intestinal segments ligation using in situ perfusion method in rats. The research demonstrated that Berberine Hydrochloride was poorly absorbed in intestine in general and mostly absorbed in the upper small intestine. The absorption fraction and absorption rate constant were gradually decreased from Duodenum to Colon. The mechanism of gastrointestinal absorption of Berberine Hydrochloride is that the drug transfers to cell via passive diffusion.
Wanshi Niuhuang Qingxin pellets with perfect shape and surface characteristics were prepared in a centrifugal granulator by using MCC as filler and 3% HPMC solution as adhesive agent under optimum condition. The dissolution rate of Berberine Hydrochloride meets the demands of preparation.
A pH-dependent gradient-release pellet system was investigated by using hydropropylated chitosan and the dispersion system of various methacrylic acid copolymers in water as coater. In vitro experiment makes it clear that various components with different properties are released synchronously.
The plasma concentration of Berberine Hydrochloride in three rabbits after a single oral administration of WSNHQX SRP was studied with WSNHQXP as a reference preparation. The results showed that the plasma concentration of Berberine Hydrochloride was steadier and its Cmax decreased significantly, A characteristic of gradient-release was observed in the concentration-time curves of Berberine Hydrochloride of WSNHQX SRP.
引文
[1] 王爱勤，贾宝全，谭干祖等．水溶性甲壳质、甲壳胺及其衍生物的制备与应用．中国海洋药物，1996，(3)：31～36
[2] 赵文伟，于黎，钟晓光等．水溶性壳聚糖的制备与应用．化学通报，1999，(4)：32～33
[3] 王振华，杜勤，许晓峰等．栀子提取工艺研究．中药材，2000，23(4)：224～226
[4] 李其兰，王国珍，卢喜珍等．用正交实验法优选小儿退热贴中栀子的提取工艺．中药材，2001，24(4)：288～290
[5] 李立荣，夏用平．TLC及柱层析分离栀子甙的比较．中成药，1999，21(12)：645～646
[6] 刘燕，吕高荣，鲁静．栀子中环烯醚萜甙成分的HPLC含量测定．中国中药杂志，1993，18(5)：298～299
[7] 徐以亮，李松，徐海波等．高效液相色谱法测定栀子浓缩颗粒中栀子苷的含量．时珍国医国药，2000，11(10)：881～882
[8] 刘乃强，江云丽，张村等．双黄连含片黄芩含量测定方法的改进．中国药学杂志，2000，35(12)：835～836
[9] 王凤有，乔艳．HPLC测定复方黄芩耳用滴丸中黄芩苷的含量．天津医科大学学报，1999，5(1)：79～80
[10] 王永林，兰燕宇，周宝珠．HPLC测定妇舒液中黄芩苷含量．中国药学杂志，1999，34(4)：265～266
[11] 王进，居文政，郁锡峰．复方薤白胶囊中盐酸小檗碱含量测定．时珍国医国药，
2001，12(1)：37～38
[12] 吴剑峰，李海燕，杨安平等．高效液相色谱法分析黄连炮制品中盐酸小檗碱的含量变化．中成药，1999，21(8)：408～409
[13] 金佩芬，缪华蓉，钱亚萍等．高效液相色谱法测定泻毒散中盐酸小檗碱的含量．中国中医药科技，2001，8(2)：102
[14] Ozdemir N, Ordu S. Improvement of dissolution properties of furosemide by complexation with beta-cyclodextrin. Drug Dev Ind Pharm,1998,24 (1), 19～25
[15] 陆彬主编．药物新剂型与新技术．第一版．北京：人民卫生出版社，1980．235～237
[16] Shobham Sabnis, Lawrence H Block.Improved infrared Spectroscopic Method for the Analysis of Degree of N-deacetylation of Chitosan.Polymer Bulletion, 1997, 39, 67～71
[17] 黄晓佳，王爱勤，袁光谱．铜离子模板法合成6-0-2-羟丙基壳聚糖．合成化学，1999，7(3)：229～231
[18] 韩蕴华．壳聚糖的羟丙基化研究．天津师范大学学报(自然科学版)，2001，21(2)：229
[19] 张千弘，马思忠，郭玉芬等．羟丙基壳聚糖的研制和鉴定．天津化工，2000，3：15～16
[20] Sannan, Shimotakaido, Suginami et al. Hydroxyalkylchitosan salt, production process thereof and cosmetic composition containing the same. European Patent Office, 1993-08-31, 585～885
[21] 王爱勤，谭干祖，贾宝全等．羟丙基壳聚糖的制备与表征．天然产物研究与开发，1997，9(1)：33～36
[22] 王爱勤，季生福．水溶性羟丙基甲壳素的合成与性能研究．天然产物研究与开
发，1995，7(3)：79～81
[23] 林瑞洵，蒋苏洪，张慕珊．脱乙酰度测定方法．化学通报，1992，5：39～42
[24] 陈鲁生，周武，姜云生．壳聚糖粘均分子量的测定．化学通报，1996，4：57
[25] Ghobad G. Maghami, George A F. Roberts.Markromol.Chem.1988, 195-189
[26] 中华人民共和国药典委员会．中华人民共和国药典．北京：化学工业出版社，2000，附录180
[27] 中华人民共和国药典委员会．中华人民共和国药典．北京：化学工业出版社，2000，附录178
[28] Lee V. Univ. Microfilms, Ann. Arbor. 1974:29446
[29] Roberts G A, Domszy J G. Int. J. Biol. Macromol. 1982, 4:347
[30] 中华人民共和国药典委员会．中华人民共和国药典．北京：化学工业出版社，2000，附录 180
[31] 申竹芳，谢明智．高效薄层荧光光密度法测定生物样品中的小檗碱含量．药学学报，1993，28(7)：532～536
[32] 安昊，王文彬．~3H-盐酸小檗碱的吸收、分布和排泄的初步研究．北京中医学院学报，1983，2：13～15
[33] 盛美萍，孙洪，王宏．盐酸小檗碱在狗静脉注射和口服药动学研究．中国药理学通报，1993，9(1)：64～67
[34] 奚念朱主编．药剂学第二版．人民卫生出版社，1994，535～540
[35] Lawrence XY, Lipka E, Crison JR, et al. Transpot approaches to the biopharmaceutical design of oral drug delivery system:prediction of intestine absorption. 1996, 19:359
[36] Stewart Ba, Chan OH, Jezyk N, et al. Descrimination between drug candidates using models for evaluation of intestinal absorption. Adv Drug Del Rev,
1997，23：275
[37] 黄志宏，方积乾主编．数理统计方法．人民卫生出版社，1987，143～147
[38] 杨正管，朱家璧，岗艳云．盐酸地尔硫卓大鼠肠吸收动力学及在口服缓释制剂设计中的意义．中国药科大学学报，1998，29(3)：179
[39] Rashid HA, Heinamaki J, Antikainen O, et al. Effect of process variables on the size, shape and surface characteristic of microcrystalline cellulose beads prepared in a centrifugal granulator. Drug Dev Ind Pharm, 1999, 25(5): 605～611
[40] 金红花，张恒弼．缓释微丸制剂研究近况．药学实践杂志，1996，14(2)：103～107
[41] 陈庆华．现代微丸制备与包衣装置的类型与应用．中国医药工业杂志，1996，27(3)：134～139
[42] 于少云，王洪光，刘璐等．微丸的进展．中国新药杂志，1999，8(12)：802～806
[43] Frank W. Goodhart. Centrifugal Equipment. Pharmaceutical Pelletization Technology. Marcel Dekker, New York, 1989, p101
[44] Nesbitt RV, Majid Mahjour, Nancy L. Mills, et al. Effect of substrate on mass release latex coated pellets. J Control Release,, 1994(32): 71
[45] 朴尤奎．离心造粒机的半自动论．中国药学杂志，1990，25(4)：34～37
[46] 陈心，隋惠卿，张芝菊．BZJ-360M型包衣造粒机在中药微丸研制中的应用．中成药，1994，16(1)：49
[47] Rashid HA, Heinamaki J, Antikainen O, et al. Effect of process variables on the size, shape and surface characteristic of microcrystalline cellulose beads prepared in a centrifugal granulator. Drug Dev Ind Pharm, 1999, 25(5): 605～611
[48] 毕殿洲主编．药剂学第四版．人民卫生出版社，1999，77～78
[49] Nakahara. US Patent 3, 277, 520. October 1966
[50] Evans DF, Pye G, Bramley R, et al. Measurement of gastrointestinal pH profiles in normal ambulant human subjects. Gut, 1988, 29: 1035～1041
[51] Davis SS, Hardy JG, Taylor MJ, et al. A comparative study of the gastrointestinal transit of a pellet and tablet formulation. Int J Pharm, 1984, 21:167～177
[52] Khan MZ, Prebeg Z, Kurjakovic N. A. pH-dependent colon targeted oral drug delivery system using methacrylic acid copolymers.Ⅰ.Manipulation of drug release using Eudragit L100-55 and Eudragit S100 combinations. Contr Rel, 1999, 58(2): 215—222
[53] Khan MZ, Stedul HP, Kurjakovic N.A pH-dependent colon targeted oral drug delivery system using methacrylic acid copolymers. Ⅱ. Manipulation of drug release using Eudragit L100 and Eudragit S100 combinations. Drug Dev Ind Pharm, 2000, 26(5): 549—554
[54] Kislalioglu MS, Khan MA, Blount C et al. Physical characterization and dissolution properties of ibuprofen: Eudragit coprecipitates. Pharm Sci, 1991, 80(8): 799—804