“内异消”缓释胶囊的研制及其药物代谢动力学研究

“内异消”缓释胶囊的研制及其药物代谢动力学研究 “内异消”缓释胶囊的研制及其药物代谢动力学研究 目录 摘 要„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„(1 Abstract(„(„„„„„(„„„„„„„((„„„((„„(„„„„„„„„„„„„(„„„„((„„„((„„„((„„IIl 英汉缩略语名词对 照„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„V1 前 言„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„(1 第一章“内异消”缓释胶囊制备工艺研 究„„„„„„„„„„„„„„„„„„„„„„4 第一节“内异消“药物含量测定方法及体外释放方法的建立„„„„„„„„„„„„„6 1实验材 料„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„6 1(1药品与试 剂„„„„„„„„„„„„„„„„„„„„„„„„„„„6 1(2实验仪器„„„„„„„„„„„„„„„„„„„„„„„„„„„„6 2实验方法与结 果„„„„„„„„„„„„„„„„„„„„„„„„„„„一6 2(1HPLC检测三种药物的含 量„„„„„„„„„„„„„„„„„„„„6 2(2紫外测定B药的含量„„„„„„„„„„„„„„„„„„„„„„(13 2(3释放度条件的确 立„„„„„„„„„„„„„„„„„„„„„„„((14 3，J、结„„„„„„„„„„„(„„„„„„„„(„„„„„„„„(„„„„„((„„„„„(„„„((15 第二节“内异消”缓释胶囊辅料的筛 选„„„„„„„„„„„„„„„„„„„„„16 1实验材 料„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„(16 6 11实验药品、试剂及辅 料„„„„„„„„„„„„„„„„„„„„„((1 1(2实验仪 器„„„„„„„„„„„„„„„„„„„„„„„„„„„(16 7 2实验方 法„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„(1 2(1胶囊的制备方 法„„„„„„„„„„„„„„„„„„„„„„„„((17 7 2(2辅料的筛 选„„„„„„„„„„„„„„„„„„„„„„„„„„((1 3，J、结„„„(„„„„„„„„(„„„„„„„„„„„„„„„„„„(„„„„„„„„„„„„„23 第三节“内异消”缓释胶囊制剂的处方优 化„„„„„„„„„„„„„„„„„„„(24 1实验材 料„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„(24 1(】实验药品、试剂及辅 料„„„„„„„„„„„„„„„„„„„„„((24 1-2实验仪 器„„„„„„„„„„„„„„„„„„„„„„„„„(’„„(24 2实验方法与结 果„„„„„„„„„„„„„„„„„„„„„„„„„„„24 2(1制备方 法„„„„„„„„„„„„„„„„„„„„„„„„„„„((24 2(2单因素考 察„„„„„„„„„„„„„„„„„„„„„„„„„„((25 2(3Box(Behenken 设计 领导形象设计圆作业设计ao工艺污水处理厂设计附属工程施工组织设计清扫机器人结构设计 „„„„„„„„„„„„„„„„„„„„„„„(35 3小 结„„„„„(„„„„„„„„(„„„„„„„„„„„„„„„(„„„(44 本章讨 论„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„46 第二章“内异消”缓释胶囊质量 标准 excel标准偏差excel标准偏差函数exl标准差函数国标检验抽样标准表免费下载红头文件格式标准下载 的研 究„„„„„„„„„„„„„„„„„„„„„48 第一节“内异消”缓释胶囊质量控制相关项目的考 察„„„„„„„„„„„„„„„49 1实验材 料„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„(49 1(1药品与试 剂„„„„„„„„„„„„„„„„„„„„„„„„„„„49 1(2实验仪 器„„„„„„„„„„„„„„„„„„„„„„„„„„„49 2实验方法与结 果„„„„„„„„„„„„„„„„„„„„„„„„„„„(49 49 2(1性状„„ 2(2鉴别„„ 50 50 2(3检查„„ 50 2(4含量测定 2(5有关物质检 查„„„„„„„„„„„„„„„„„„„„„„„„„(51 1 2(6释放度„„„„„„„„„„„„„„„„„„„„„„„„„„„„(5 3小结„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„54 4(讨j仑„„„„„„„„„((„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„(54 第二节“内异消”缓释胶囊质量标准 草案 及起草说明„„„„„„„„„„„„„„55 第三章“内异消”缓释胶囊的稳定性研 究„„„„„„„„„„„„„„„„„„„„„„59 】实验材 料„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„(59 1(1药品与试 剂„„„„„„„„„„„„„„„„„„„„„„„„„„(59 1(2(实验仪 器„„„„„„„„„„„„„„„„„„„„„„„„„„„59 2实验方法与结 果„„„„„„„„„„„„„„„„„„„„„„„„„„„(59 2(1影响因素实 验„„„„„„„„„„„„„„„„„„„„„„„„„((60 2-2加速实 验„„„„„„„„„„„„„„„„„„„„„„„„„„„((65 2-3长期实 验„„„„„„„„„„„„„„„„„„„„„„„„„„„((69 3小结„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„ „„„„„„„„„(69 4讨1仑„„(„„(„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„69 第四章“内异消’罐爱释胶囊药物代谢动力学的研 究„„„„„„„„„„„„„„„„„„(70 1实验材 料„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„(70 1(1药品与试 剂„„„„„„„„„„„„„„„„„„„„„„„„„„70 1(2实验仪 器„„„„„„„„„„„„„„„„„„„„„„„„„„„((70 1(3实验动 物„„„„„„„„„„„„„„„„„„„„„„„„„„„(7l 2实验方法与结 果„„„„„„„„„„„„„„„„„„„„„„„„„„„(7l 2(1 HPLC检测大鼠血浆中三个药物的含 量„„„„„„„„„„„„„„„7l 2(2大鼠体内药动学研 究„„„„„„„„„„„„„„„„„„„„„„，76 3小结„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„80 4讨j仑(„„„„„„„„„„„(„„„„„„„„„„„„„„„„„„„„„„„„„„„„(„„((80 全文总 结„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„8l 创新及意 义„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„82 思考与展 望„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„83 参考文 献„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„84 文献综 述„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„86 致 谢„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„„(97 攻读硕士学位期间科研工作汇 报„„„„„„„„„„„„„„„„„„„„„„„„„98 摘要 “内异消"缓释胶囊的研制及其药物代谢动力学研究 研究生-唐勤 硕士专业:药物分析学 指导教师:徐晓玉教授 摘要 目的: 制备“内异消”缓释胶囊，使其药物组分阿魏酸、盐酸川芎嗪、延胡索 乙素 三药均能达到缓释效果，并从质量标准、稳定性和体内缓释性方面进行评价。 方法: 1(?内异消彦缓释胶囊的制各工艺研究考察亲水凝胶缓释辅料H、不溶性缓 释辅料E及不同溶蚀性辅料 O、M、S、G 对阿魏酸、盐酸川芎嗪、延胡索乙 素三种药物的缓释效果，筛选最佳的辅料和制备方法;在单因素考察的基础上， h、6h、12 采用Box(Behenken设计，以复方中三个药物2h的累积释放的综合评 分为指标，对药物辅料比 蜀 、两辅料的比例 弱 、颗粒过筛目数 x3 进行优 化，最终确定“内异消”缓释胶囊的制备方法。 2(。内异消”缓释胶囊质量标准研究按照药典的相关要求结合本制剂特点， 从性状、装量差异、粒径、有关物质、鉴别、含量测定、释放度等方面建立“内 异消”缓释胶囊的质量标准，拟定质量标准 草案 。 3(搿内异消一缓释胶囊的稳定性研究对“内异消”缓释胶囊进行影响因素 实验、加速实验和长期稳定性实验，考察其稳定性。 4(“内异消’’缓释胶囊药物代谢动力学的研究采用高效液相色谱法建立了大 鼠血浆样品中“内异消”缓释胶囊各组分的含量测定方法:大鼠灌胃给予原料药 和缓释胶囊内容物 180 mg??kg’1 ，定时取血，检测各时间点血浆中的药物浓度。 DAS软件计算药代动力学参数，考察缓释胶囊在大鼠体内的缓释性能。 结果: T 西南大学硕士论文 1(“内异消一缓释胶囊制备工艺研究确定“内异消”缓释胶囊的制备方法为: 药物与辅料的比例为X，两辅料H和G的比例为Y，将药物与辅料H混匀后;0hA( 到熔融的辅料G中，搅拌，混匀，冷却30rain，过Z目筛，装胶囊。 2(“内异消”缓释胶囊质量标准研究 1 含量测定方法:甲醇(0(5,醋酸 42:58 为流动相，检测波长280 nllq，三药的含量均在标示量的90,110,之间。 2 装量差异 士7(5,: 3 能通过80目和未通过60目粒径的质量小于总质量 的10,; 4 药物阿魏酸在2h、6h、12h的的释放量应满足10,20,、60,70,、 90,100,;盐酸川I芎嗪在2h、6h、12 h的的释放量应满足35--45,、60,70,、 85,95,; h、6h、12 延胡索乙素在2 h的的释放量应满足20,30,，55---65,， 80-90,。 5 有关物质采用主成分阿魏酸稀释25倍对照法，小于对照溶液中阿 魏酸面积的5,; 3(“内异消一缓释胶囊稳定性研究影响因素实验中湿度对胶囊的影响 最大; 加速实验3个月的结果显示“内异消”缓释胶囊的含量、有关物质及释放度相对 稳定。 4(“内异消一缓释胶囊药物代谢动力学研究 1 以香豆素为内标，流动相 甲醇(0(5,醋酸 38:62 ，检测波长280nm，建立的体内含量测定方法，该方法 稳定，系统适应性好。 2 大鼠血浆药物代谢动力学研究表明:胶囊内容物中阿 魏酸、盐酸川芎嗪、延胡索乙素三种药物的半衰期是原料药的2(3l、’2(53、2(62倍， 达峰时间分别从0(52h延长至3(25h，从O(46 h， h延长至2(5h，从0(69h延长至5 表明胶囊缓释效果明显:药时曲线下面积分别是原料药的2(1、1(25和1(85倍，药 物的生物利用度有所提高。 结论: 以H和G为缓释辅料，熔融制粒法制备“内异消”缓释胶囊，工艺操作简单， 质量易控，重复性好。制得的缓释胶囊体内外均具有良好的缓释性能，提高了药 物生物利用度。 关键词:子宫内膜异位症:缓释:熔融制粒法;阿魏酸;盐酸川芎嗪;延胡索 乙素 Il 英汉缩略语名词对照 of studies The and Neiyixiao pharmacokinetic preparation Sustained-release capsule Candidate:TangQin Pharmaceutical analysis Major: ProfessorXu Supervisor: Xiao-yu Abstract Objective ferulic 1"o amethodthatcaninsurethe establish acid， hydrochloricligustrazine effectand the toreachsusutainedrelease and in tetrahydropalmatineprescription invitroandinvivo( thesusutainedreleaseeffect evaluate Methods Theeffectsofdifferent of sustained―release Neiyixiao capsules 1(Preparation sustainedrelease sustainedrelease aboutwater-soluble H，insoluble excipient excipients suchas andGwere ECanderodible 0，M(S investigated; Though excipients excipients modelwasestablishedto the factors Box??Behenken the optimize experiments，A single 12hofthree in were releaseof2 hand factos，thecumulative h，6 drugNeiyixiao index( optimization tothe sustained-release controlof capsulesAccording 2(Quality Neiyixiao size， relatedsubstances， difference，particle content，loading Pharmacopoeia，the the ofthe were tocontrol identification capsules， thequality investigated quality wasestablished draft sustained??re]ease standards capsule ofNeiyixiao TheSustained。 release of sustained??release capsules 3(StabilityNeiyixiao andthe accelerated weretakenunderthefactor experiment experiment，the capsule to the ofthe studystabilityproduct long(termexperiment inratsThe of sustained―release 4(Pharmacokinetics capsules Neiyixiao 111 西南大学硕士论文 ratswere methoddetectthethreesubstancesinvivowereestablished HPLC;The by doseof1 administratedofofbulk and atthe intragastric drugscapsule 80mg,kg， the wastestedHPLCtodetectthe atdifferenttimeafteradministration(The by drug plasma were DASsoftwere( pharmacokineticparametersanalysisedby Results method of sustained-releaseThe Neiyixiao capsulespreparation 1(Preparation arex(HandGare andHwere wasestablished:Atthe and xcipients Y(thedrug drug with werecooledafter30 addedtothemoltenG stirring，thecompound accompanyed mininroom and temperature，sievedcapsuled controland of sustained-release 2(Quality stabilityNeiyixiao capsules 1 At ofmethanol-0(5,acetic themobile nmas thecondition acid 42:58 as phase，280 ferulic and detection contentof wavelength(The acid， hydrochloricligustrazine atthe90-110,astothelabeled wereall Tetrahydropalmatine amount 2 Load sizewascontrolledwithin90, between60mesh difference 士7(5,( 3 Theparticle and80 cumulativedreleaseofferulicacidin2 2hshouldmeet h，6h，1 mesh( 4 The off 10,20,、60-70,、90MOO,:Theacumulativedrelease ligustrazine in2 2hshouldmeetthe h，6h，1 35,45,，60,70,，85, 95,;The hydrochloride in 2 2 h shouldmeet acumulativedreleaseof h，6h，1 tetrahydropalmatine 20-30,，55-65,，80-90,( of sustained-release resultsshowedthat 3(StabilityNeiyixiao capsulesStability threefactors(It’S wasthe factors the content， related humidity importantamong intheaccelerated inthree substancesandreleasewere stable experiment relatively month sustained??releasein the 3(Pharmacokineticsof Neiyixiao rats: 1 At capsules conditionofmethanol??0(5,acetic themobile amas acid 38:62 asphase，280 detection andCoumarinasinternal effective standard，the wavelength drug ingredients allhada liner methodwas forthedeterminationthemin good good relationchip，the witha and the in experiments plasmagoodprecionrecovery( 2 Frompharmacokinetic IV 英汉缩略语名词对照 offerulic and were rats，thet1,2 acid， hydrochloricligustrazineTetrahydropalmatine 2(31、2(53、2(62timesastobuik ofthethree wereextendfromthe T drugs;the drugs to the0(46hto2(5 theAUCwere2(1、 0(52hto3(25h(fromthe0(46 2(5h(from h(and 1(25和1(85timstothebulkdrugs( Conclusion: The Sustained―release melt was granulation Neiyixiao capsulepreparedby proved tohaveasustainedeffectinvitroandin the controland vivo，and quality stability a and tocontrol it’S easy experimentprovedsimple preparation( fusion words:Endometriosis;Sustained??release;hotmethod; Ferulicacid， Key Hydrochloricligustrazine;Tetrahydropalmatin V 西南大学硕士论文 英汉缩略语名词对照 中文名称 英文缩写 英文名称 阿魏酸 A ferulicacid curve 药时曲线下面积 AUC Areaunder 盐酸川芎嗪 B ligustrazine hydrochloric 延胡索乙素 C tetrahydropalmatine ofvariation 变异系数 CV toefficient 清除率 CL Clear 天 d day similarfactors 相似因子 五 克 gram g hour 小 时 h 高 效液相色谱 HPLC chromatogram highperformanceliquid 盐 酸 HCL acid Hydrochloric 千 克 kilogram k 分钟 mln mmute month 月 m 毫米 mm millimeter molar 摩尔 mol 毫克 milligram mg M1 milliliter 毫升 Mole 摩尔 M residencetime 平均滞留时间 MRT mean 纳克 Nanogram ng 纳米 nm nanometer buffers 磷酸盐缓冲液 PBS Phosphate 聚乙烯吡咯烷酮 PVP Polyvinylpyrrolidone relativestandarddeviation 相对标准偏差 RSD 相对湿度 RH Relative humidity mlcrogram 微克 “g microlitre 微 升 uL 微米 micrometer 肛m ? 摄氏度 temperature VI 前言 ―jI―(L 刖吾 1(子宫内膜异位症缺乏有效的治疗药物 体和间质 出现在子宫腔以外的其他地方 主要是女性的内脏和腹腔壁上 ，是一 种妇科良性疾病，但种植的子宫内膜具有浸润、扩散等恶性行为，常伴有慢性骨 盆的疼痛、痛经、月经量增多、慢性盆腔包块、不孕等症状，给女性的生活带来 了极大的不便。据统计，子宫内膜异位症在育龄妇女中发病率高达10,以上，并 且随着年龄的增长和病程的延长，异位种植的子宫内膜细胞具有癌变的倾向【1之]。 因此，上世纪九十年代本病被世界卫生组织 WHO 列为疑难疾病。 目前治疗EMs的方法有手术和药物治疗，手术治疗有腹腔镜检异味组 织切除 手术，但需要考虑病人的对生育的态度。而药物治疗主要采用激素治疗，包括口 服避孕药类、促性腺激素类、雄性激素类以及孕激素类，但这些药物都存在不同 程度的副作用，如乳房胀痛、体重增加、粉刺、闭经、妇女多毛以及骨质疏松等， 并且在停止用药后容易复发?治疗效果均不令人满意。 2(中药治疗子宫内膜异位症有其特有的优势 祖国医学典籍中类似于本病的记载很多，对其病因病机、诊治等皆有论述， 如《金匿要略》中有:“经水不利，少腹满痛„”的记述。《医林改错》中用少腹 逐瘀汤治疗“少腹积块疼痛，或有积块不疼痛，或疼痛而无积块”等。中医 多根 据辨病辨症相结合的原则，用补肾祛瘀、清热化瘀、活血化瘀、活血通腑、温阳 活血及补肾固冲等方法治疗本病。其病症主要表现为内膜周期性出血，为“离经 出血”，或称为蓄血或淤血;其病位在下焦，血瘀是本病的基本病理因素，活血化 瘀是治疗基本法则，再根据疼痛的部位、性质、及伴随症状、舌象、脉象结合病 史诊合，寻求血瘀的成因，采取辨证施治，在改善疼痛、缓解症状、提高受孕率 以及减轻毒副作用等方面有较好的优势【钔。现代中医学家也普遍认为本病的根本病 机是血瘀，1990年中国中西医结合学会妇产科专业委员会第三届学术会议将子宫 内膜异位症中医诊断标准修订为血瘀证。西医研究认为子宫内膜异位症是由多因 素相互影响形成的结果，这恰与中药多靶点、整体治疗相吻合。 l 西南大学硕士论文 随着对EMs研究的深入，我国中医药界利用传统中医药的特长和优势，在治 疗子宫内膜异位症方面做了不少的努力和探讨，发现了许多有效的复方‘51，并积累 了丰富的经验，取得了较为满意的疗效"】。 但中药的传统剂型一一水煮煎剂在长 适合长期使用的中成药新剂型对治疗 期的应用中比较困难，故开发有效、方便、 本病具有重要的意义。 3(中药缓控释制剂适用于慢性疾病的治疗 中药的缓释思想在古医书上中早有记载，“丸者，缓也，不能速去之，其用药 之舒缓而治之意也”[81，但传统丸剂只是缓释制剂雏形，其缓释行为具有模糊性， 不是现代意义上的缓释制剂。现代意义的缓控释制剂是指通过恰当的方法，延缓 药物在体内的吸收、分布和代谢的过程，使药物在体内的血药浓度控制在治疗窗 范围内。目的是在提高药物疗效、降低毒副作用的同时，减少用药次数，提高患 者用药顺应性。 西药在缓释制剂方面已目趋成熟，中药缓释制剂是在遵循中医理论的基础 上，借助西药缓释制剂药动学设计、辅料应用、成型工艺、生物药剂学特性、质 量评价等发面的理论，在考虑原处方中各成分相互作用及疾病需要的同时，合理 发展的中药剂型【91。但中药及其复方具有多成分、多作用、多途径、多靶点的整合 调节作用，它又不同于结构清楚、成分单一、作用靶点明确的化学药品，这些体 现中药缓释制剂的复杂性。 缓释制剂在治疗需长期服药的慢性疾病患者方面有独特的优势，可减少用药 次数，提高患者的顺应性[10]。子宫内膜以为症是一种妇科慢性疾病，因此在药物 的治疗上采用缓释制剂更适合疾病的发病特点，同时提高患者的顺应性。 4(“内异消一治疗子宫内膜异位症有比较好的疗效 “内异消”是由课题组自主配方、用于治疗子宫内膜异位症的中药复方制剂， 是由中药著名方剂佛手散中当归和JII芎的主要药效成分阿魏酸 A 和盐酸川芎嗪 B 加上具有止痛作用的延胡索乙素 C 三种药物按照一定比例组成。课题组 近年来对其主要药效学进行了考察111-]2]，表明3种药物联合使用对治疗子宫内膜 异位症具有良好的效果，没有显著毒副反应，已经申报国家技术发明专利，并得 ， 前言 到科技部“重大新药创帛0”重庆孵化基地专项资助。但药物要作用于机体，必须 制备成适当的剂型，因此将药物制备成一定的剂型对于新药来说显得如此重要。 而缓释制剂对慢性疾病的病人可提高其顺应性。 基于以上背景，本课题欲将对子宫内膜异位症有很好治疗作用的中药复方药 物“内异消”开发为缓释胶囊，旨在能更加适应疾病的发病特点和提高患者顺应 性。为建立中药复方缓释制剂对理化性质差异较大的各组分实现缓释的手段和评 价方法提供参考。 西南大学硕士论文 第一章“内异消"缓释胶囊制备工艺研究 复方“内异消”中含有三种主要药效成分包括阿魏酸、盐酸川芎嗪和延胡索 乙素。通过文献调研，对三药的性质考察如下: 是植物中广泛分布的一种芳香酸，有顺式和反式两种 结构如图1(】一1A所示 ， 溶点为174。C，溶于热水、乙醇和乙酸乙酯，稍溶于乙醚，难溶予苯和石油醚。 阿魏酸溶液不稳定。研究表明，阿魏酸溶液对光照极为敏感，在强光照射下会 部分转化为其顺式产物;随温度的增加，降解速率增加;酸性和碱性环境下降 解速率增加，而在中性环境下最为稳定【】31。 且不易溶于水，为了增加原料药的稳定性，因此药典中收载的是川芎嗪的盐酸盐 和磷酸盐的两种形式。该药为我国所特有，未见国外药典收载及相关产品上市。 实验室以盐酸川芎嗪作为原料药，为白色或类白色结晶性粉末，气味特异，味苦， 熔点85,90。C，在水中易溶，在乙醇或氯仿中溶解，在苯中极微溶解 结构如图 1-1(1B所示 。盐酸川芎嗪的药代动力学特点是半衰期短 t1，2 2(89士0(5h ，目前 临床广泛的用于血管疾病，作用确切，毒副作用小[141。但是临床的静脉滴注给药 后，血药浓度迅速下降，而口服缓释制剂成了近年来广大研究者的热点。水溶性 药物扩散性强，骨架片释放面积较大，往往会存在突释现象[】5]，因此，对于水溶 性药物若能在控制其突释的同时又保证其释药完全，就可以得到优良的缓释制剂。 延胡索乙素是从中药延胡索中提取的一种生物碱，为消旋体，其有效部位为 左旋体，即左旋四氢巴马汀 结构如图1(1(1C所示 ，该药物呈淡黄色结晶状， 熔点148。C116]。它是新中国成立以来，用现代科学技术研究中医药获得成功的第一 个神经系统药物。大量的有关研究证实延胡索乙素除用于镇痛、镇静、安定外， 还是治疗阵发性房颤、快速型室上性心律失常安全有效的药物，并对顽固性呃逆、 剧烈咳嗽有显著疗效?7J。 内异消缓释胶囊制备工艺研究 ( C A B (p Ha??2H，O 0、( H0 0, 图1(1(1阿魏酸 A 、盐酸川芎嗪 B 、延胡索乙素 C 的结构式 1-1-1StructureofFerulic Figure acid A ，ligustrazinehydrochloride B and tetrahydropalmatine C 三个单体药物理化性质差异大，阿魏酸为酸性物质，盐酸川芎嗪为强酸弱碱 盐，延胡索乙素为生物碱;从溶解度来看，三种物种在不同pH环境中的溶解差异 大。这些差异为在同一处方中使三药达到同步缓释带来了难题。并且阿魏酸的溶 液不稳定，盐酸川芎嗪的熔点较低，这些药物本身的特点是选择方法的时候应该 注意的。 本章通过体外筛选不同的辅料，确定制备技术和方法，并通过处方的优 化， 将复方“内异消”研制成为缓释胶囊， 西南大学硕士论文 第一节“内异消"药物含量测定方法及体外释放方法的建立 1实验材料 1(1药品与试剂 氢氧化钠 成都市科龙化工试剂厂 磷酸二氢钠 成都市科龙化工试剂厂 浓盐酸 成都市科龙化工试剂厂 甲醇 色谱纯 阿达马斯公司 冰醋酸 色谱纯 成都市科龙化工试剂厂 纯化水 Millipore纯水系统 阿魏酸对照品 批号:110817-201003 中国药品生物制品检定所 盐酸川芎嗪对照品 批号:110773(201004 中国药品生物制品检定所 延胡索乙素对照品 批号:110726(201002 中国药品生物制品检定所 1(2实验仪器 200高效液相色谱色谱仪 美国安捷伦公司 Agilentl FA2004A电子天平 上海精工电子仪器有限 公司 SHB(IllS循环水式多用真空泵 郑州长城科工贸有限公司 D(800LS智能药物溶出仪 天大天发溶出科技有限公司 U3010紫外分光光度计 日立高新科技有限公司 2实验方法与结果 2(1 HPLC检测三种药物的含量 2(1(1检测波长的确立 对三种药物紫外图谱扫描，确定最佳的检测波长。 内异消缓释胶囊制备工艺研究 A B C 瑚25。 3D。 瑚如0 wave'lengltl』nm 图1(1(2A、B、C三药的紫外扫描图谱 l一1-2The UV ofA，B，C Figur scanningspectra by 如图1(1(2所示，药物A、B、C在280nm均有最大吸收，确定A、B、 C三 物质的检测波长为280nm。 2(1(2色谱条件 通过文献的查阅【181及预实验，确定色谱条件。其色谱条件如下: SB―C18ODS色谱柱 250×4(6 mm:5岬 ; 色谱柱:Agilent 流动相:甲醇(0(5,的醋酸 52:48; 流速:O(8mL(rain,: 柱温:30?; 检测波长:280nm; 进样量:10”L; 2(1(3系统适用性试验 照2(1(2项下的色谱，将原料药和制剂溶于甲醇中，超声溶解，其 图谱如图 1(1(3所示: 7 西南大学硕士论文 A 2 b B C min 1 4 6 7 8 9 1 O 2 3 5 0 2 3 4 S 6 7 8 9 min 图1(1(3“内异消”中药物对照品 a 及制剂 b 色谱图 1-1-3The ofreference materials a and Figure chromatogram preparations b in Neiyixiao min出峰。各组分 分 如图1(1(3所示，辅料对测定无干扰，溶剂与辅料在前3 min，C与A的分离度为2(7， 离度好，A、B、C的保留时间分别为5(2、7(3、3(7 A与B之间的分离度为3(3，三种物质的理论塔板数分别为2500，2345，2400。 2(1(4线性相关实验 mL棕色容量 分别精密称取A、B、C三药19(9 mg、14(6mg、11(4mg于25 瓶中，加入一定量的甲醇，超声溶解，冷却，定容。分别量取1、0(5、O(2、O(1、 O(OlmL于10mL容量瓶中，甲醇定容至刻度。照2(1(2项下的条件进行 分析，各 药物的浓度与峰面积结果如表1(1-1、1(1―2、1(1(3所示。 表1(1(1不同浓度A药的峰面积 concentrationofA Table1(1(1Peakareaofdiffbrent 内异消缓释胶囊制备工艺研究 表1(1(3不同浓度C药的峰面积 Table1-1―3PeakareaofdifferentconcentrationofC 照上表结果，得到三种药物的标准曲线如图1(1(4所示: ? 簟 0_ 图1(1-4A、B、C三药的标准曲线 1―1-4Standardcu?eofA(BandC Figure 将对所得标准曲线进行线性回归，得到线性方程如表l―1(4所示: 表1―1-4A、B、C三种药物的回归方程及线性范围 Table1-1―4 andthelinear oftheA、B、C Regressionequation range 回归方程 相关系数 R 线性范围 ”g??mL’1 该条件下r A、B、C三药在0(796,79(6岭??mL,，B药在0(584,58(4gg??mL,、 C药在0(456-45(6 gg??mL。之间线性关系良好。 2(1(5精密度实验 取2(1(4项下A成分线性范围内高 39(80L(1 、中 15(92mL‘1 、低 gg??m gg 7(96 lag。mL。 三个浓度，B成分线性范围内高 29(20 gg??mL‘1 、中 11(68 日 西南大学硕士论文 嵋‘mL一 、低 5(84 Pg‘mL一 三个浓度，C成分线性范围内高 22(80 gg(mL一 、 中 9(12 I(tg‘mL叫 、低 4，561(tg‘mL。 三个浓度，日内进样5次，考察目内精密 度;日间进样5天，考察日间精密度;根据标准曲线测得浓度，得到日内 RSD和 日间RSD。照2(1(2项下的色谱条件进行分析，测定药物的浓度，其结 果如表1(1(5、 1(】一6、1(】(7所示: 表1一1(5 A药的日内精密度和日间精密度 i士如，n 5 Tablel??1―5 and ofA fi士(如(n 51 Intra-dayprecisioninter-dayprecision 日内精密度 日间精密度 RSD , RSD , 浓度 pg-mL‘1 浓度 gg??mL’1 表1-1-6 B药的日内精密度和日问精密度 Y。+SD。刀 5 and precisioninter-dayprecisionofB i士(妨，n--5 ―Tablel―-1-6Intm―-d―ay 表1(1-7 C药的日内精密度和日间精密度 y。-4-SD，n 5 Table1-l-7 and ofC fi士肋(n 5 Intra―dayprecisioninter-dayprecision 日内精密度 日间精密度 RSD , RSD , 浓度 gg??mL。1 浓度 gg??mL‘1 如上表所示，该方法在的日内精密度在0(08?4(04,之间，小于5,， 曰间 精密度在1(85,,7(11,之间，小于10,，可用于三种物质的检测。 10 2(1(6回收率实验 样品溶液的制备:精密称取胶囊内容物适量，加入适量甲醇，超声溶解，冷 1(70 mL，测定其中各药物的含量 A为23(00pg’mL,，B为1 却至室温，定容至100 mL，按照线性实验高、中、低浓度 I(tg-mL‘1 。移取样品溶液1 Llg'mL,，C为7(10 mL 于10mL容量瓶中，甲醇定容。照2。1，2 分别加入不同体积 O(5、0(2、0(1 项下色谱条件项进行分析，测定药物的浓度。三药的回收率结果如表l??1-8、卜1-9、 1(1―10所示: 表1(1(8A药的回收率 (