超微如意金黄散巴布膏剂的制备及其体外透皮吸收作用与主要药效、临床疗效的研究
|
摘要
[目的]
选择经典名方“如意金黄散”为研究载体,对其超微粉与普通粉进行选择试验,通过两者主要成份的溶出度及粉体特征与薄层色谱、体外透皮吸收作用及主要药效学等进行比较试验,探索微粉化对外用散剂主要有效成分溶出速率与体外透皮吸收作用与主要药效的影响,探讨中药外用散剂的适宜粒径,为超微粉体技术应用于中药外用散剂提供参考；对超微如意金黄散的制备工艺进行系统的研究,制定其规范化的工艺示范,建立其内在质量与外在质量的评价指标,为超微如意金黄散巴布膏剂的制备提供前期工作试验基础；采用正交试验,对超微如意金黄散巴布膏剂的制剂处方进行了筛选,建立了适用于规模化生产的巴布膏剂生产工艺,并初步探讨了超微如意金黄散巴布膏剂治疗急性软组织损伤早期之气滞血瘀证的临床疗效,为经典名方“如意金黄散”的深度开发奠定基础。
[方法]
1、如意金黄散超微粉与其普通粉的选择：应用超微粉体技术,处方各药味选择饮片为原料,以盐酸小檗碱与姜黄素溶出度、姜黄素体外透皮吸收速率等为指标,对如意金黄散超微粉与其普通粉进行比较,优选粉体粒径。
1.1溶出度试验：参照《中国药典》2010版二部附录XC溶出度测定法,以盐酸小檗碱、姜黄素为指标,采用桨法,检测两种不同粒径粉体的溶出度,考察两种粉体的溶出特性,优选适宜粒径的粉体。
1.2体外透皮吸收度试验：以姜黄素为指标,考察不同粉体粒径对如意金黄散主要有效成分透皮速率的影响,探索有效成分姜黄素累积透过量与粉体粒径的关系。
1.3显微特征的比较试验：分别取如意金黄散超微粉与其普通粉适量,用水合氯醛透化,置显微镜下观察并拍照,比较两者显微特征的差别。
1.4薄层色谱的比较试验：对处方中的大黄、白芷、姜黄等药味进行薄层色谱比较试验,在取样量相同、处理方法一致的前提下,比较超微如意金黄散与其普通散的薄层色谱差异。
2超微如意金黄散制备工艺的研究
2.1通过对单独粉碎与混合粉碎、振动粉碎与气流粉碎进行比较,确定如意金黄散的超微粉碎工艺。
2.2以盐酸小檗碱溶出量为指标,选择适宜的粉碎粒径。
2.3采用单因素试验方法,对如意金黄散超微粉碎的入磨物料水分、入磨粒度、介质填充率、投入量、粉碎时间等工艺参数进行考察,确定超微如意金黄散的最佳工艺技术参数。
3质量控制指标及方法的研究
3.1选择性状、粉体表征、比表面积、堆密度、休止角及粒径等作为超微如意金黄散的外在质量评价指标
3.2以水分、理化性质、薄层鉴别、含量测定、溶出度等为其内在质量评价指标。
3.3建立了超微如意金黄散的质量标准。
4、超微如意金黄散巴布膏剂的制备
4.1采用正交试验及综合评分法,以聚丙烯酸钠、酒石酸、甘羟铝、CMC-Na、甘油-山梨醇(70%)为因素,设置四水平的L16(45)正交设计试验,优选如意金黄散巴布膏剂成型工艺的制剂配方。
4.2通过中试生产工艺验证,考察生产工艺的稳定性。
5、主要药效的研究
5.1镇痛作用：采用扭体法,考察超微如意金黄散与其普通散、超微如意金黄散巴布膏剂与其普通巴布膏剂,对小鼠醋酸扭体反应的影响,比较其镇痛作用。
5.2抗炎作用：采用二甲苯致炎模型,考察超微如意金黄散与其普通散、超微如意金黄散巴布膏剂与其普通散巴布膏剂对小鼠耳廓二甲苯致炎的影响,比较其抗炎作用。
6、超微如意金黄散巴布膏治疗急性软组织损伤临床疗效的观察：采用随机、安慰剂平行对照、双盲法,观察超微如意金黄散巴布膏治疗急性软组织损伤的疗效,并对超微如意金黄散巴布膏临床用药的安全性进行考察。
[结果]
1如意金黄散超微粉与其普通粉的选择
1.1溶出度试验：如意金黄散超微粉体与其普通粉体中盐酸小檗碱溶出量分别为0.0971g和0.0926g。
1.2体外透皮吸收试验：以姜黄素为指标,建立的高效液相含量测定法重复性好、方法稳定；如意金黄散超微粉与其普通粉中所含姜黄素可以透皮,其透过量随时间的延长而增加,透过量与时间有较好的线性关系；如意金黄散超微粉在给药第3h时,兔皮上姜黄素的累积渗透量达25.22μg/cm2,而其普通粉在给药第3h时,兔皮上姜黄素的累积渗透量仅达17.37μg/cm2。
1.3显微特征：如意金黄散普通粉能明显观察到无色团块状物,纤维单个散在,石细胞大多成群,也有单个散离的,形状及大小不一,草酸钙针晶细小,散在于射线细胞中,油细胞随处可见；而其超微粉中药材的显微特征不明显,虽然也能观察到石细胞,但绝大多数已破损,纤维细胞成碎片。
1.4薄层色谱：在取样量、样品处理方法相同的条件下,如意金黄散的超微粉与其普通粉的薄层色谱中,分别在与大黄、白芷、姜黄对照药材和姜黄素对照品相应的位置上,均显示相同颜色的斑点,且其超微粉样品的斑点较普通粉更清晰明显。
2、超微粉碎工艺的研究
2.1通过单独粉碎与混合粉碎、振动粉碎与气流粉碎进行比较,确定如意金黄散采用混合粉碎,且振动粉碎优于气流粉碎。
2.2主要有效成分溶出量的实验结果表明,如意金黄散的适宜粉碎粒径为D9()36μm。
2.3采用单因素试验方法,确定如意金黄散的工艺参数为：贝利机粉碎,振幅为5.5mm,粉碎温度为0～5℃,入磨物料水分为3～6%、入磨粒度为粗粉,介质填充率为60～70%,投料量为250g,振动时间为30mmin。
3超微如意金黄散的质量标准研究：根据3批超微如意金黄散的样品的性状描述、粉体表征、比表面积、堆密度、休止角及粒径检测等,拟定了超微如意金黄散的外在质量评价指标,根据3批超微如意金黄散样品的水分、理化性质、薄层鉴别、含量测定、溶出度,拟定了其内在质量评价指标,建立了超微如意金黄散的可控质量标准。
4、如意金黄散巴布膏剂的制备
4.1优选出超微如意金黄散巴布剂的基质配方,即巴布剂基质组成为：聚丙烯酸钠、酒石酸、甘羟铝、CMC-Na、甘油、山梨醇,配比为30：1：0.8：7：100：80,能够制备弹性好、粘性强,易于敷贴的巴布膏剂。
4.2三批中试验证结果表明,拟定的工艺可行,产品质量稳定。
5、如意金黄散巴布膏剂主要药效的研究
5.1镇痛作用：如意金黄散超微粉与其普通粉、如意金黄散超微粉与其普通粉的巴布膏剂均能降低醋酸所致小鼠扭体反应次数,具较好的镇痛作用。且如意金黄散超微粉及其巴布膏剂的镇痛作用明显优于其普通粉及其巴布膏剂。
5.2抗炎作用：如意金黄散超微粉与其普通粉、如意金黄散超微粉与其普通粉的巴布膏剂均能明显抑制二甲苯所致小鼠耳肿胀,具有明显的抗炎消肿作用。且如意金黄散超微粉及其巴布膏剂的抗炎作用明显优于其普通粉及其巴布膏剂。
6、超微如意金黄散巴布膏的临床疗效观察
临床疗效观察结果表明,超微如意金黄散巴布膏剂治疗急性软组织损伤的总显效率为80%,总有效率为90.00%,对照组(伤湿祛痛膏)的总显效率为23.33%,总有效率为53.33%；超微如意金黄散巴布膏对气滞血瘀证的总显效率为86.67%,有总效率96.67%,对照组(伤湿祛痛膏)总显效率为23.33%,总有效率为53.33%。病、证疗效组间比较差异均有统计学意义,治疗组明显优于对照组。在试验过程中,治疗组有1例病例敷贴局部皮肤出现轻度小皮疹过敏现象,停用药物未经处理自愈,没有观察到其他明显不良反应。本次临床试验两组受试者用药前后生命指征、安全性指标均无具有明显临床意义的变化。
[结论]
1.采用现代超微粉体技术,使“超微如意金黄散”粉体的平均粒径达0.1～75μm,促进了有效成分的溶出,与普通散比较,超微粉中姜黄素、盐酸小檗碱的溶出速率快,起效迅速。
2.以姜黄素为指标的体外透皮吸收试验表明,超微如意金黄散中的姜黄素透皮速率较其普通散快速,提示,超微粉体制剂有提高生物利用度的优势,具有开发应用前景。
3.薄层色谱表明,超微如意金黄散样品中大黄、姜黄、白芷等药味的主斑点较其普通散样品更清晰、明显,提示,微粉化促进了其有效成分的溶出,为超微粉体技术在外用散剂中的应用提供了试验依据。
4.成型工艺研究表明,采用正交试验优选的超微如意金黄散巴布剂基质配方利于成型；中试研究表明,工艺稳定,质量稳定。
5.镇痛、抗炎试验结果表明,如意金黄散超微粉及其巴布膏剂的镇痛、抗炎作用均明显优于其普通粉及其巴布膏剂。
6.临床疗效观察结果表明,超微如意金黄散巴布膏剂治疗急性软组织损伤的总显效率为80%,总有效率为90.00%,安全性较好。
7.本论文研究结果为超微如意金黄散巴布膏剂的开发奠定了坚实的基础,可带来较大的社会、经济效益
Objective
Taking Ruyijinhuangsan as the research object, Based on studying of the pharmaceutical technology, characteristic, TLC, Pharmacodynamics, and Transdermal Absorption in vitro about the ultra fine and ordinary powder with Ruyijinhuangsan, In order to explore the advantages of ultra-fine powder about the Extracting of Effective Components and Pharmacodynamics in compound. To offer Scientific information for dose conversion about the ultra fine and ordinary powder. Optimizing the best formula of hydrophilic cataplasm, in order to establish a set of Cataplasm Techniques with the advantages of general practicality.
Methods
1Ruyijinhuangsan ultrafine and its choice of powder:application of superfine powder technology, Selectting Pieces of the prescription for the raw of various ingredients materials. Using hydrochloric berberine, curcumin dissolution of curcumin in vitro transdermal absorption rate as an indicator, this paper wish to compare its ultrafine and ordinary powder and Prefere the particle size of its.
1.1Dissolution test:reference "Chinese Pharmacopoeia2010edition two appendix XC dissolution determination method, berberine, curcumin as indicators, using the paddle method, the detection of two different particle size powders dissolution investigated two kinds of powderdissolution characteristics of the body, preferably suitable for the powder of particle size.
1.2Transdermal absorption test:Using Curcumin as an indicator to study the impact of the different particle size wishful golden powder of the active ingredient in transdermal rate.Exploring the active ingredient curcumin accumulated through the amount of powder particle size.
1.3Comparison of the microscopic character about ultra fine and ordinary powder, Adopting chloral hydrate method and using microscope.
1.4studying on the TLC of Rhubarb, Angelica and Turmeric, Comparison of the TLC about the ultra fine and ordinary powder in ruyijinhuangsan, by the premise of consistent with the sampling amount and method.
2Ruyijinhuangsan powder preparation process
2.1Comparing Crushed separately,mixed vibration crushed and the air smash,Used it to determine the Ruyijinhuangsan powder ultrafine grinding process.
2.2Using Berberine hydrochloride dissolution as the indicators, to choice the appropriate of crushed particle size.
2.3single factor test, Using the ground water, particle size, medium fill rate, inputs into the mill, grinding process parameters as indicators, To determine the best technical parameters of Advanced Micro Devices, Ruyijinhuang powder.
3Quality control indicators and methods
3.1Select traits, powder characterization, surface area, bulk density, angle of repose and particle size as the external quality evaluation of Supermicro ruyijinhuangsan.
3.2Using water, dissolution, physicochemical properties, TLC, and determination as its intrinsic quality evaluation.
3.3Establishing Supermicro Ruyijinhuangsan powder quality standards.
4Preparation of ultra-fine Ruyijinhuang powder Cataplasm Transdermal.
4.1Adopted carbomer sodium polyacrylate, CMC-Na, Glycerol, tartaric acid and dihydroxyaluminum aminoacetate as indicators, using L16(45) orthogonal design in order to Optimizing cataplasma matrix.
4.2Through pilot production, process validation to study the stability of the production process.
5The main pharmacodynamic study
5.1Analgesic effect:With using Kunming mouse(SPF) as the experimental animal, in order to study on the Influence of jasminoidin on inflammation of pinnae in mice,and also the different of effects on Pharmacodynamics.
5.2Anti-inflammatory effects:With using Kunming mouse(SPF) as the experimental animal, in order to study on the Influence of acetic acid torsion reaction in mice.
660patients with Acute soft tissue injury were divided into two groups randomly:theraphy group (ultra Ruyijinhuang Cataplasm Transdermal)、 contrasted group (Injury wet cured pain cream). With pain, swelling, tenderness, ecchymosis, dysfunction, dry mouth, constipation, tongue, pulse other grading score and disappear recovery time as factors to calculate the efficacy of change.
Results
1The choice of Ruyijinhuangsan superfine and ordinary powder.
1.1Dissolution testing:Superfine powder and its berberine hydrochloride in dissolution of the ordinary powder were0.0971g and0.0926g.
1.2The HPLC method is stable and reliable, The ultra fine powder and ordinary powder of Ruyijinhuansan contained curcumin transdermal increased through the extension of the amount of time, and has a good linear relationship between amount of time; The amount iontophoretic flux reached25.22μg/cm2,while Ruyijinhuansan ultra fine powder on the administration of the first3h in rabbit, but the ordinary powder of Ruyijinhuansan only17.37μg/cm2.
1.3Ordinary powder can be clearly observed to a colorless mass tilting, fiber single scattered, mostly in groups of stone cells, there are also single scattered from the shape and sizes. Small calcium oxalate crystals scattered in ray cells, oil cells can be seen everywhere; and ultra fine powder characteristics of the microstructure is not obvious, although it can be observed to the stone cells, but the vast majority of damaged fiber cell debris, and it is no complete oil cells to see.
1.4When the same conditions as sample volume, sample handling, The ultra fine powder show spots more clear, ultrafinedarker and obvious than ordinary powder of Rhubarb, Angelica and Turmeric, on TLC.
2Ultrafine grinding process
2.1Selectting Separately,mixed grinding vibration crushed and air jet mill, to determine the RuYiJinHuangSan mixing crushed, and the vibration crush is better than air jet mill.
2.2The dissolution of active ingredients show that Ruyijinhuangsan powder suitable for crushed particle size D9036μm.
2.3Using Single factor test to determine the process parameters Ruyijinhuangsan powder:Billy machine grinding, the amplitude of5.5mm, smash a temperature of0-5℃into the grinding material water for3to6%into the grinding particle size for the mealmedium fill rate of60to70percent, the feeding amount of250g, vibration time of30min.
3Advanced Micro Devices, Ruyijinhuangsan powder quality standard:Describe the three batches of ultrafine powder samples, using powder characterization, specific surface area, bulk density, angle of repose and particle size detection, and prepared external quality evaluation of Advanced Micro Devices, Ruyijinhuangsan powder, according to the moisture of the three batches of ultrafine powder samples, dissolution, physicochemical properties, TLC identification, assay, developed its intrinsic quality evaluation to establish the quality standards of Advanced Micro Devices, Ruyijinhuangsan powder.
4Preparation of ultra-fine Ruyijinhuang powder Cataplasm Transdermal.
4.1Optimized cataplasma:the indicators formulation are matrix carbomer sodium polyacrylate, CMC-Na, Glycerol, tartaric acid and dihydroxyaluminum aminoacetate:30:1:0.8:7:100:80.
4.2it was verified for three batch of the pilot test, so this technology is feasible and stable product quality.
5The main pharmacodynamic study
5.1Analgesic effect:With using Kunming mouse(SPF) as the experimental animal, in order to study on the Influence of jasminoidin on inflammation of pinnae in mice,and also the different of effects on Pharmacodynamics.
5.2Anti-inflammatory effects:With using Kunming mouse(SPF) as the experimental animal, in order to study on the Influence of acetic acid torsion reaction in mice,and also the different of effects on Pharmacodynamics.
6The treatment of acute soft tissue injury markedly effective rate in the Supermicro Ruyijinhuangsan Papua paste was80%, the total efficiency of90.00%, in the control group were23.33%and53.33%. It is total markedly effective rate was86.67%, the over all efficiency of96.67%in the control group were23.33%and53.33%. Disease, card between the efficacy of group differences were statistically significant, the treatment group was significantly better than the control group.
Conclusion
1Using modern ultrafine grinding technology to the Ruyijinhuansan powder to be the average particle size of0.1～75μm, The microscopic characteristics of after ultrafine herbs powder is significantly reduced, uniform in size, and the oil cells to be broken, the active ingredients of essential oils can be better exposed, rather than through the transparent wall (membrane) to release, So the medicinal efficacy can release faster from herbs.
2Using curcumin as an indicator of in vitro percutaneous absorption test, the transdermal rate of curcumin in Supermicro Ruyijinhuansan powder faster than ordinary power, suggesting that the ultrafine powder formulations have certain advantages, and has prospects for the development and application.
3The ultra fine powder show spots more clear, obvious than ordinary powder of Rhubarb, Angelica and Turmeric, on TLC, That preparations made of ultrafine active ingredients more stripping, providing a reference for the improvement of the dosage form and dosage.
4The Supermicro Ruyijinhuangsan powder cataplasm matrix formula conducive to molding, the quality of finished product is stability, and mainly pharmacodynamic exact.
5In this thesis, the results provide new agents for Supermicro Ruyijinhuang powder cataplasm and it will bringing larger social and economic benefits.
6Clinical observation of the treatment about acute soft tissue injury with Ultra-fine Ruyijinhuangsan Cataplasm, it provides a clinical basis for the development of new formulation.
选择经典名方“如意金黄散”为研究载体,对其超微粉与普通粉进行选择试验,通过两者主要成份的溶出度及粉体特征与薄层色谱、体外透皮吸收作用及主要药效学等进行比较试验,探索微粉化对外用散剂主要有效成分溶出速率与体外透皮吸收作用与主要药效的影响,探讨中药外用散剂的适宜粒径,为超微粉体技术应用于中药外用散剂提供参考；对超微如意金黄散的制备工艺进行系统的研究,制定其规范化的工艺示范,建立其内在质量与外在质量的评价指标,为超微如意金黄散巴布膏剂的制备提供前期工作试验基础；采用正交试验,对超微如意金黄散巴布膏剂的制剂处方进行了筛选,建立了适用于规模化生产的巴布膏剂生产工艺,并初步探讨了超微如意金黄散巴布膏剂治疗急性软组织损伤早期之气滞血瘀证的临床疗效,为经典名方“如意金黄散”的深度开发奠定基础。
[方法]
1、如意金黄散超微粉与其普通粉的选择：应用超微粉体技术,处方各药味选择饮片为原料,以盐酸小檗碱与姜黄素溶出度、姜黄素体外透皮吸收速率等为指标,对如意金黄散超微粉与其普通粉进行比较,优选粉体粒径。
1.1溶出度试验：参照《中国药典》2010版二部附录XC溶出度测定法,以盐酸小檗碱、姜黄素为指标,采用桨法,检测两种不同粒径粉体的溶出度,考察两种粉体的溶出特性,优选适宜粒径的粉体。
1.2体外透皮吸收度试验：以姜黄素为指标,考察不同粉体粒径对如意金黄散主要有效成分透皮速率的影响,探索有效成分姜黄素累积透过量与粉体粒径的关系。
1.3显微特征的比较试验：分别取如意金黄散超微粉与其普通粉适量,用水合氯醛透化,置显微镜下观察并拍照,比较两者显微特征的差别。
1.4薄层色谱的比较试验：对处方中的大黄、白芷、姜黄等药味进行薄层色谱比较试验,在取样量相同、处理方法一致的前提下,比较超微如意金黄散与其普通散的薄层色谱差异。
2超微如意金黄散制备工艺的研究
2.1通过对单独粉碎与混合粉碎、振动粉碎与气流粉碎进行比较,确定如意金黄散的超微粉碎工艺。
2.2以盐酸小檗碱溶出量为指标,选择适宜的粉碎粒径。
2.3采用单因素试验方法,对如意金黄散超微粉碎的入磨物料水分、入磨粒度、介质填充率、投入量、粉碎时间等工艺参数进行考察,确定超微如意金黄散的最佳工艺技术参数。
3质量控制指标及方法的研究
3.1选择性状、粉体表征、比表面积、堆密度、休止角及粒径等作为超微如意金黄散的外在质量评价指标
3.2以水分、理化性质、薄层鉴别、含量测定、溶出度等为其内在质量评价指标。
3.3建立了超微如意金黄散的质量标准。
4、超微如意金黄散巴布膏剂的制备
4.1采用正交试验及综合评分法,以聚丙烯酸钠、酒石酸、甘羟铝、CMC-Na、甘油-山梨醇(70%)为因素,设置四水平的L16(45)正交设计试验,优选如意金黄散巴布膏剂成型工艺的制剂配方。
4.2通过中试生产工艺验证,考察生产工艺的稳定性。
5、主要药效的研究
5.1镇痛作用：采用扭体法,考察超微如意金黄散与其普通散、超微如意金黄散巴布膏剂与其普通巴布膏剂,对小鼠醋酸扭体反应的影响,比较其镇痛作用。
5.2抗炎作用：采用二甲苯致炎模型,考察超微如意金黄散与其普通散、超微如意金黄散巴布膏剂与其普通散巴布膏剂对小鼠耳廓二甲苯致炎的影响,比较其抗炎作用。
6、超微如意金黄散巴布膏治疗急性软组织损伤临床疗效的观察：采用随机、安慰剂平行对照、双盲法,观察超微如意金黄散巴布膏治疗急性软组织损伤的疗效,并对超微如意金黄散巴布膏临床用药的安全性进行考察。
[结果]
1如意金黄散超微粉与其普通粉的选择
1.1溶出度试验：如意金黄散超微粉体与其普通粉体中盐酸小檗碱溶出量分别为0.0971g和0.0926g。
1.2体外透皮吸收试验：以姜黄素为指标,建立的高效液相含量测定法重复性好、方法稳定；如意金黄散超微粉与其普通粉中所含姜黄素可以透皮,其透过量随时间的延长而增加,透过量与时间有较好的线性关系；如意金黄散超微粉在给药第3h时,兔皮上姜黄素的累积渗透量达25.22μg/cm2,而其普通粉在给药第3h时,兔皮上姜黄素的累积渗透量仅达17.37μg/cm2。
1.3显微特征：如意金黄散普通粉能明显观察到无色团块状物,纤维单个散在,石细胞大多成群,也有单个散离的,形状及大小不一,草酸钙针晶细小,散在于射线细胞中,油细胞随处可见；而其超微粉中药材的显微特征不明显,虽然也能观察到石细胞,但绝大多数已破损,纤维细胞成碎片。
1.4薄层色谱：在取样量、样品处理方法相同的条件下,如意金黄散的超微粉与其普通粉的薄层色谱中,分别在与大黄、白芷、姜黄对照药材和姜黄素对照品相应的位置上,均显示相同颜色的斑点,且其超微粉样品的斑点较普通粉更清晰明显。
2、超微粉碎工艺的研究
2.1通过单独粉碎与混合粉碎、振动粉碎与气流粉碎进行比较,确定如意金黄散采用混合粉碎,且振动粉碎优于气流粉碎。
2.2主要有效成分溶出量的实验结果表明,如意金黄散的适宜粉碎粒径为D9()36μm。
2.3采用单因素试验方法,确定如意金黄散的工艺参数为：贝利机粉碎,振幅为5.5mm,粉碎温度为0～5℃,入磨物料水分为3～6%、入磨粒度为粗粉,介质填充率为60～70%,投料量为250g,振动时间为30mmin。
3超微如意金黄散的质量标准研究：根据3批超微如意金黄散的样品的性状描述、粉体表征、比表面积、堆密度、休止角及粒径检测等,拟定了超微如意金黄散的外在质量评价指标,根据3批超微如意金黄散样品的水分、理化性质、薄层鉴别、含量测定、溶出度,拟定了其内在质量评价指标,建立了超微如意金黄散的可控质量标准。
4、如意金黄散巴布膏剂的制备
4.1优选出超微如意金黄散巴布剂的基质配方,即巴布剂基质组成为：聚丙烯酸钠、酒石酸、甘羟铝、CMC-Na、甘油、山梨醇,配比为30：1：0.8：7：100：80,能够制备弹性好、粘性强,易于敷贴的巴布膏剂。
4.2三批中试验证结果表明,拟定的工艺可行,产品质量稳定。
5、如意金黄散巴布膏剂主要药效的研究
5.1镇痛作用：如意金黄散超微粉与其普通粉、如意金黄散超微粉与其普通粉的巴布膏剂均能降低醋酸所致小鼠扭体反应次数,具较好的镇痛作用。且如意金黄散超微粉及其巴布膏剂的镇痛作用明显优于其普通粉及其巴布膏剂。
5.2抗炎作用：如意金黄散超微粉与其普通粉、如意金黄散超微粉与其普通粉的巴布膏剂均能明显抑制二甲苯所致小鼠耳肿胀,具有明显的抗炎消肿作用。且如意金黄散超微粉及其巴布膏剂的抗炎作用明显优于其普通粉及其巴布膏剂。
6、超微如意金黄散巴布膏的临床疗效观察
临床疗效观察结果表明,超微如意金黄散巴布膏剂治疗急性软组织损伤的总显效率为80%,总有效率为90.00%,对照组(伤湿祛痛膏)的总显效率为23.33%,总有效率为53.33%；超微如意金黄散巴布膏对气滞血瘀证的总显效率为86.67%,有总效率96.67%,对照组(伤湿祛痛膏)总显效率为23.33%,总有效率为53.33%。病、证疗效组间比较差异均有统计学意义,治疗组明显优于对照组。在试验过程中,治疗组有1例病例敷贴局部皮肤出现轻度小皮疹过敏现象,停用药物未经处理自愈,没有观察到其他明显不良反应。本次临床试验两组受试者用药前后生命指征、安全性指标均无具有明显临床意义的变化。
[结论]
1.采用现代超微粉体技术,使“超微如意金黄散”粉体的平均粒径达0.1～75μm,促进了有效成分的溶出,与普通散比较,超微粉中姜黄素、盐酸小檗碱的溶出速率快,起效迅速。
2.以姜黄素为指标的体外透皮吸收试验表明,超微如意金黄散中的姜黄素透皮速率较其普通散快速,提示,超微粉体制剂有提高生物利用度的优势,具有开发应用前景。
3.薄层色谱表明,超微如意金黄散样品中大黄、姜黄、白芷等药味的主斑点较其普通散样品更清晰、明显,提示,微粉化促进了其有效成分的溶出,为超微粉体技术在外用散剂中的应用提供了试验依据。
4.成型工艺研究表明,采用正交试验优选的超微如意金黄散巴布剂基质配方利于成型；中试研究表明,工艺稳定,质量稳定。
5.镇痛、抗炎试验结果表明,如意金黄散超微粉及其巴布膏剂的镇痛、抗炎作用均明显优于其普通粉及其巴布膏剂。
6.临床疗效观察结果表明,超微如意金黄散巴布膏剂治疗急性软组织损伤的总显效率为80%,总有效率为90.00%,安全性较好。
7.本论文研究结果为超微如意金黄散巴布膏剂的开发奠定了坚实的基础,可带来较大的社会、经济效益
Objective
Taking Ruyijinhuangsan as the research object, Based on studying of the pharmaceutical technology, characteristic, TLC, Pharmacodynamics, and Transdermal Absorption in vitro about the ultra fine and ordinary powder with Ruyijinhuangsan, In order to explore the advantages of ultra-fine powder about the Extracting of Effective Components and Pharmacodynamics in compound. To offer Scientific information for dose conversion about the ultra fine and ordinary powder. Optimizing the best formula of hydrophilic cataplasm, in order to establish a set of Cataplasm Techniques with the advantages of general practicality.
Methods
1Ruyijinhuangsan ultrafine and its choice of powder:application of superfine powder technology, Selectting Pieces of the prescription for the raw of various ingredients materials. Using hydrochloric berberine, curcumin dissolution of curcumin in vitro transdermal absorption rate as an indicator, this paper wish to compare its ultrafine and ordinary powder and Prefere the particle size of its.
1.1Dissolution test:reference "Chinese Pharmacopoeia2010edition two appendix XC dissolution determination method, berberine, curcumin as indicators, using the paddle method, the detection of two different particle size powders dissolution investigated two kinds of powderdissolution characteristics of the body, preferably suitable for the powder of particle size.
1.2Transdermal absorption test:Using Curcumin as an indicator to study the impact of the different particle size wishful golden powder of the active ingredient in transdermal rate.Exploring the active ingredient curcumin accumulated through the amount of powder particle size.
1.3Comparison of the microscopic character about ultra fine and ordinary powder, Adopting chloral hydrate method and using microscope.
1.4studying on the TLC of Rhubarb, Angelica and Turmeric, Comparison of the TLC about the ultra fine and ordinary powder in ruyijinhuangsan, by the premise of consistent with the sampling amount and method.
2Ruyijinhuangsan powder preparation process
2.1Comparing Crushed separately,mixed vibration crushed and the air smash,Used it to determine the Ruyijinhuangsan powder ultrafine grinding process.
2.2Using Berberine hydrochloride dissolution as the indicators, to choice the appropriate of crushed particle size.
2.3single factor test, Using the ground water, particle size, medium fill rate, inputs into the mill, grinding process parameters as indicators, To determine the best technical parameters of Advanced Micro Devices, Ruyijinhuang powder.
3Quality control indicators and methods
3.1Select traits, powder characterization, surface area, bulk density, angle of repose and particle size as the external quality evaluation of Supermicro ruyijinhuangsan.
3.2Using water, dissolution, physicochemical properties, TLC, and determination as its intrinsic quality evaluation.
3.3Establishing Supermicro Ruyijinhuangsan powder quality standards.
4Preparation of ultra-fine Ruyijinhuang powder Cataplasm Transdermal.
4.1Adopted carbomer sodium polyacrylate, CMC-Na, Glycerol, tartaric acid and dihydroxyaluminum aminoacetate as indicators, using L16(45) orthogonal design in order to Optimizing cataplasma matrix.
4.2Through pilot production, process validation to study the stability of the production process.
5The main pharmacodynamic study
5.1Analgesic effect:With using Kunming mouse(SPF) as the experimental animal, in order to study on the Influence of jasminoidin on inflammation of pinnae in mice,and also the different of effects on Pharmacodynamics.
5.2Anti-inflammatory effects:With using Kunming mouse(SPF) as the experimental animal, in order to study on the Influence of acetic acid torsion reaction in mice.
660patients with Acute soft tissue injury were divided into two groups randomly:theraphy group (ultra Ruyijinhuang Cataplasm Transdermal)、 contrasted group (Injury wet cured pain cream). With pain, swelling, tenderness, ecchymosis, dysfunction, dry mouth, constipation, tongue, pulse other grading score and disappear recovery time as factors to calculate the efficacy of change.
Results
1The choice of Ruyijinhuangsan superfine and ordinary powder.
1.1Dissolution testing:Superfine powder and its berberine hydrochloride in dissolution of the ordinary powder were0.0971g and0.0926g.
1.2The HPLC method is stable and reliable, The ultra fine powder and ordinary powder of Ruyijinhuansan contained curcumin transdermal increased through the extension of the amount of time, and has a good linear relationship between amount of time; The amount iontophoretic flux reached25.22μg/cm2,while Ruyijinhuansan ultra fine powder on the administration of the first3h in rabbit, but the ordinary powder of Ruyijinhuansan only17.37μg/cm2.
1.3Ordinary powder can be clearly observed to a colorless mass tilting, fiber single scattered, mostly in groups of stone cells, there are also single scattered from the shape and sizes. Small calcium oxalate crystals scattered in ray cells, oil cells can be seen everywhere; and ultra fine powder characteristics of the microstructure is not obvious, although it can be observed to the stone cells, but the vast majority of damaged fiber cell debris, and it is no complete oil cells to see.
1.4When the same conditions as sample volume, sample handling, The ultra fine powder show spots more clear, ultrafinedarker and obvious than ordinary powder of Rhubarb, Angelica and Turmeric, on TLC.
2Ultrafine grinding process
2.1Selectting Separately,mixed grinding vibration crushed and air jet mill, to determine the RuYiJinHuangSan mixing crushed, and the vibration crush is better than air jet mill.
2.2The dissolution of active ingredients show that Ruyijinhuangsan powder suitable for crushed particle size D9036μm.
2.3Using Single factor test to determine the process parameters Ruyijinhuangsan powder:Billy machine grinding, the amplitude of5.5mm, smash a temperature of0-5℃into the grinding material water for3to6%into the grinding particle size for the mealmedium fill rate of60to70percent, the feeding amount of250g, vibration time of30min.
3Advanced Micro Devices, Ruyijinhuangsan powder quality standard:Describe the three batches of ultrafine powder samples, using powder characterization, specific surface area, bulk density, angle of repose and particle size detection, and prepared external quality evaluation of Advanced Micro Devices, Ruyijinhuangsan powder, according to the moisture of the three batches of ultrafine powder samples, dissolution, physicochemical properties, TLC identification, assay, developed its intrinsic quality evaluation to establish the quality standards of Advanced Micro Devices, Ruyijinhuangsan powder.
4Preparation of ultra-fine Ruyijinhuang powder Cataplasm Transdermal.
4.1Optimized cataplasma:the indicators formulation are matrix carbomer sodium polyacrylate, CMC-Na, Glycerol, tartaric acid and dihydroxyaluminum aminoacetate:30:1:0.8:7:100:80.
4.2it was verified for three batch of the pilot test, so this technology is feasible and stable product quality.
5The main pharmacodynamic study
5.1Analgesic effect:With using Kunming mouse(SPF) as the experimental animal, in order to study on the Influence of jasminoidin on inflammation of pinnae in mice,and also the different of effects on Pharmacodynamics.
5.2Anti-inflammatory effects:With using Kunming mouse(SPF) as the experimental animal, in order to study on the Influence of acetic acid torsion reaction in mice,and also the different of effects on Pharmacodynamics.
6The treatment of acute soft tissue injury markedly effective rate in the Supermicro Ruyijinhuangsan Papua paste was80%, the total efficiency of90.00%, in the control group were23.33%and53.33%. It is total markedly effective rate was86.67%, the over all efficiency of96.67%in the control group were23.33%and53.33%. Disease, card between the efficacy of group differences were statistically significant, the treatment group was significantly better than the control group.
Conclusion
1Using modern ultrafine grinding technology to the Ruyijinhuansan powder to be the average particle size of0.1～75μm, The microscopic characteristics of after ultrafine herbs powder is significantly reduced, uniform in size, and the oil cells to be broken, the active ingredients of essential oils can be better exposed, rather than through the transparent wall (membrane) to release, So the medicinal efficacy can release faster from herbs.
2Using curcumin as an indicator of in vitro percutaneous absorption test, the transdermal rate of curcumin in Supermicro Ruyijinhuansan powder faster than ordinary power, suggesting that the ultrafine powder formulations have certain advantages, and has prospects for the development and application.
3The ultra fine powder show spots more clear, obvious than ordinary powder of Rhubarb, Angelica and Turmeric, on TLC, That preparations made of ultrafine active ingredients more stripping, providing a reference for the improvement of the dosage form and dosage.
4The Supermicro Ruyijinhuangsan powder cataplasm matrix formula conducive to molding, the quality of finished product is stability, and mainly pharmacodynamic exact.
5In this thesis, the results provide new agents for Supermicro Ruyijinhuang powder cataplasm and it will bringing larger social and economic benefits.
6Clinical observation of the treatment about acute soft tissue injury with Ultra-fine Ruyijinhuangsan Cataplasm, it provides a clinical basis for the development of new formulation.
引文
[1]蔡光先.中药粉体工程学[M].北京：人民卫生出版社,2008.
[2]李富文,金风媚.超微粉碎技术在中药业中的应用[J].中国动物保健,2003(11)：35-36.
[3]邢征.从重压研磨式超微粉碎机看中药超微粉碎的新理念[J].机电信息,2005,1(16)：59
[4]杜晓敏,刘璐.原生药材超细微粉制剂的药效学研究[J].中草药,1999,30(9)：680-684..
[5]王丽娟,刘训红,宋建平，等.麋鹿角超细粉体表征及其水溶性蛋白质溶出度的研究[J].南京中医药大学学报,2010,26(2)：132.
[6]宋建平,王丽娟,刘训,等,超微粉碎技术对麇鹿角主要化学成分提取率的影响[J].时珍国医国药,2011,22(6)：1431-1433.
[7]曹龙奎,黄威,王景会,等.玉米花粉超微粉碎破壁技术的实验研究[J].农业工程学报,2003,19(6)：209-211.
[8]刘云海,杜光.超微粉碎对中药活性成分提取率的影响[J].中国医院药学杂志,2010,30(1)：66-69.
[9]苏瑞强,何煜,王瑞成.超微粉碎技术提高六味地黄丸(水蜜丸)溶出度的研究[J].2002,27(7)：511-513.
[10]谷雨,王玉蓉,杨连威.超微粉碎技术对白芷中香豆素类成分的影响[C].中华中医药学会第九届制剂学术研讨会论文汇编,2010：98-102.
[11]陈实功.外科正宗[M].北京：人民卫生出版社,1964：1.
[12]林治琳,林晓瑜.加昧如意金黄散外敷治疗骨折及软组织损伤[J].山东中医杂志,2008,27(10)：681-682.
[13]周龙恒.加味金黄散外敷治疗急性软组织损伤[J].中医正骨,2008,20(10)：68.
[14]兰绍波,宋修亭.如意金黄散外治湿疹132例[J].辽宁中医杂志,2006,33(4)：48.
[15]倪绍军,战广茂.如意金黄散治疗肛周湿疹30例[J].中国民康医学杂志,2005,17(7)：346.
[16]国家药典委员会.中华人民共和国药典[S].北京：化学工业出版社,2010：720-721.
[17]毛平,夏卉莉.如意金黄贴膏与如意金黄散消肿止痛功效的比较研究[J].中国实验方剂学杂志,2004,10(5)：46-48.
[18]刘芳,刘小平.增效如意金黄散软膏制各工艺的研究[J].湖北中医杂志,2006,28(6)：52-53.
[19]卢静华,杨欢.双波长薄层扫描法测定如意金黄散中盐酸小檗碱的含量[J].中国生化药物杂志,2011,32(1)：52-53.
[20]刘占军,韩刚.如意金黄散膜剂的改进[J].时珍国医国药,2009,20(7)：1782-1783.
[21]陈永财,周斌,等.正交试验法优选“如意金黄散”巴布剂基质配方[J].海峡药学,2008,20(9)：12-14.
[22]赵浩如,杨永刚,何煜.肉桂超细微粉的粉体特征及体外透皮作用的研究[J].中成药,2002,24(9)：654-656.
[23]严华,魏锋.同属不同种大黄及含大黄制剂中土大黄苷检查方法的研究[J].药物分析杂志,2010,(9)：1615-1620.
[24]张清.黑膏药疮疡膏工艺及质量标准研究[J].时珍国医国药,2008,19(10)：2423-2425.
[25]李明,周大颖,田园,等.中药姜黄的薄层色谱研究[J].贵州化工,2008,(1)：31-34.
[26]王静,秦伟,张全明.TLC法测定散结乳癖贴膏中姜黄素的含量[J].中国药房,2007,18(18)：1394-1395.
[27]张康,何宇新,吴丽等.正交试验优选镇痛贴乙醇提取工艺[J].中国实验方剂学杂志,2011,17(5)：37-39.
[28]杨敏,杨光琼,赵青HPLC法测定如意金黄散中姜黄素的含量[J].现代医药卫生,2011,27(10)：1461-1462.
[29]楼步青.双黄巴布剂体外透皮促进剂的筛选[J].时珍国医国药,2011,22(9)：2247-2250.
[30]吴青青,陈彦.姜黄素脂质体的制备及体外透皮研究[J].中国医药工业杂志,2011,42(12)：910-913.
[31]孙冬梅,李智勇,邓亚利.癌痛巴布剂的质量标准研究[J].江西中医学院学报, 2011,23(4)：34-37.
[32]赵美玉.浅谈巴布剂的生产工艺[J].中国中医药咨讯.2011,3(8)：288-288.
[33]刘方艺,谢友良,蒋东旭.正交设计法优化经络贴巴布剂基质配方研究[J].中国医药导报,2011，8(10)：68-70.
[34]庞来祥,寻园,郭鑫等.正交试验法优化通络止痛巴布剂配方究[J].中国民族民间医药杂志,2011,20(3)：48-49.
[35]徐华明,江国荣.金黄乳膏抗炎镇痛作用的实验研究[J].中医正骨，2011,23(9)：14-16.
[36]刘媛,王健.羊软骨Ⅱ型胶原蛋白对小鼠佐剂性关节炎抑制作用的研究[J].食品科学，2010,(15)：268-270.
[37]陈奇.中药药理研究方法学[M],人民卫生出版社,第1版,1993：378.
[1]陈实功.外科正宗[M].北京：人民卫生出版社,1964：1.
[2]国家药典委员会.中华人民共和国药典[S].北京：化学工业出版社,2010：720-721.
[3]舒林利,李莉.75%乙醇联合如意金黄散外敷治疗静脉炎[J].护理学杂志，2010，25(11)18-19.
[4]邓家英,吴仲安,符关芳.如意金黄散外敷治疗化疗性静脉炎84例[J].中国民康医学,2010,22(1)：11.
[5]林艳容.如意金黄散外敷治疗静脉留置针并发静脉炎[J].浙江中西医结合杂志,2010,20(3)：179.
[6]Bouiton A J. The diabetic foot:aglobal view[J]. Diabetes Metab R es R ev,2000,16 suppll:S2-S5.
[7]贾秀卿,孙喜明.如意金黄散外敷在糖尿病足溃疡护理中的应用[J].临床和实验医学杂志,2010,9(10)：786-787.
[8]和玉英,文歧琳.如意金黄散外敷配合抬高架减轻下肢痛风性关节炎急性期肿痛的疗效观察及护理[J].辽宁中医药大学学报,2009,11(9)：144-145.
[9]王京红.如意金黄散治疗皮肤疮疬肿毒32例[J].中国现代药物应用,2009,3(6)：78.
[10]柴维霞.如意金黄散联合神灯治疗四肢急性软组织挫伤疗效观察[J].中国中医急症,2012,21(1)：170.
[11]王晓,陈欣菊.如意金黄膏封包治疗肝硬化合并自发性腹膜炎58例明[J].中国中医药信息杂志,2007,14(12)：74.
[12]杨玉英,许增宝,朱炜,等.如意金黄散熏洗治疗慢性前列腺炎的效果观察[J].护理与康复,2005,4(2)：86-87.
[13]陈玉涛,王五俊.浓茶冲调如意金黄散外敷辅助治疗阑尾周围脓肿43例临床分析[J].黑龙江中医药,2009,(3)：24-25.
[14]Minghetti, PCasirahgiA, Giluzro, Fetal.Development of local patehes containing melilot extract and ex vivo-in vivo evaluation of skin permeation[J].Euor J Pharm Sci, 2000,10:111-117.
[15]王建明,王政超.抗风湿涂膜剂的制剂研究[J].哈尔滨商业大学学报,2011,27(5)：662-666.
[16]原素敏,倪健.脉炎宁透皮贴剂成型工艺及体外透皮释放研究[J].中国中药杂志,2009,34(13)：1654-1657.
[17]周小凤,杨中林,李萍,等.不同纯度苦杏仁普巴布剂的透皮吸收比较研究[J].药学与临床研究,200,16(4)：285-287.
[18]刘占军,韩刚.如意金黄散膜剂的改进[J].时珍国医国药,2009,20(7)：1782-1783.
[19]陈永财,周斌,等.正交试验法优选“如意金黄散”巴布剂基质配方[J].海峡药学,2008,20(9)：12-14.
[20]刘芳,刘小平.增效如意金黄散软膏制各工艺的研究[J].湖北中医杂志,2006,28(6)：52-53.
[21]韩刚,张卫国,王超,等.高效液相色谱法测定如意金黄散中5种成分的含量[J].中国医院药学杂志,2007,27(1)：124-125.
[22]毛泉明,毛平,叶福嫒,等.反相高效液相色谱法测定如意金黄贴膏中小檗碱的含量[J].中国医院药学杂志,2002,22(8)：477-478.
[23]卢静华,杨欢.双波长薄层扫描法测定如意金黄散中盐酸小檗碱的含量[J].中国生化药物杂志,2011,32(1)：52-53.
[24]江汉美,卢金清,刘璇,等.聚酰胺薄层扫描法测定如意金黄散中厚朴酚与和厚朴酚的含量[J].湖北中医学院学报,2009,11(5)：27-28.
[25]林治琳，林晓瑜.加昧如意金黄散外敷治疗骨折及软组织损伤[J].山东中医杂志,2008,27(10)：681-682.
[26]周龙恒.加味金黄散外敷治疗急性软组织损伤[J].中医正骨,2008，20(10)：68.
[27]兰绍波,宋修亭.如意金黄散外治湿疹132例[J].辽宁中医杂志,2006,33(4)：48.
[28]倪绍军,战广茂.如意金黄散治疗肛周湿疹30例[J].中国民康医学杂志,2005,17(7)：346.
[29]Godin B, Touitou E. Erythromycin ethosomal systems:physi-Cochemical Characterization and enhanced antibacterial activity [J]. Curr Drug Deliv,2005,2(3) 269-275.
[30]Dayan N,Touitou E.Carriers for skin delivery of trihexyphenidyl HCL:ethosomes vs. Liposomes [J]. Biomaterials,2000,21:1879-1885.
[31]Touitou E,Godin B.Dayan N,et al.Intracellular delivery mediated by an ethosomal carrier [J]. Biomaterials,2001,22:3053-3059.
[32]毛平,夏卉莉.如意金黄贴膏与如意金黄散消肿止痛功效的比较研究[J].中国实验方剂学杂志,2004,10(5)：46-48.
[2]李富文,金风媚.超微粉碎技术在中药业中的应用[J].中国动物保健,2003(11)：35-36.
[3]邢征.从重压研磨式超微粉碎机看中药超微粉碎的新理念[J].机电信息,2005,1(16)：59
[4]杜晓敏,刘璐.原生药材超细微粉制剂的药效学研究[J].中草药,1999,30(9)：680-684..
[5]王丽娟,刘训红,宋建平，等.麋鹿角超细粉体表征及其水溶性蛋白质溶出度的研究[J].南京中医药大学学报,2010,26(2)：132.
[6]宋建平,王丽娟,刘训,等,超微粉碎技术对麇鹿角主要化学成分提取率的影响[J].时珍国医国药,2011,22(6)：1431-1433.
[7]曹龙奎,黄威,王景会,等.玉米花粉超微粉碎破壁技术的实验研究[J].农业工程学报,2003,19(6)：209-211.
[8]刘云海,杜光.超微粉碎对中药活性成分提取率的影响[J].中国医院药学杂志,2010,30(1)：66-69.
[9]苏瑞强,何煜,王瑞成.超微粉碎技术提高六味地黄丸(水蜜丸)溶出度的研究[J].2002,27(7)：511-513.
[10]谷雨,王玉蓉,杨连威.超微粉碎技术对白芷中香豆素类成分的影响[C].中华中医药学会第九届制剂学术研讨会论文汇编,2010：98-102.
[11]陈实功.外科正宗[M].北京：人民卫生出版社,1964：1.
[12]林治琳,林晓瑜.加昧如意金黄散外敷治疗骨折及软组织损伤[J].山东中医杂志,2008,27(10)：681-682.
[13]周龙恒.加味金黄散外敷治疗急性软组织损伤[J].中医正骨,2008,20(10)：68.
[14]兰绍波,宋修亭.如意金黄散外治湿疹132例[J].辽宁中医杂志,2006,33(4)：48.
[15]倪绍军,战广茂.如意金黄散治疗肛周湿疹30例[J].中国民康医学杂志,2005,17(7)：346.
[16]国家药典委员会.中华人民共和国药典[S].北京：化学工业出版社,2010：720-721.
[17]毛平,夏卉莉.如意金黄贴膏与如意金黄散消肿止痛功效的比较研究[J].中国实验方剂学杂志,2004,10(5)：46-48.
[18]刘芳,刘小平.增效如意金黄散软膏制各工艺的研究[J].湖北中医杂志,2006,28(6)：52-53.
[19]卢静华,杨欢.双波长薄层扫描法测定如意金黄散中盐酸小檗碱的含量[J].中国生化药物杂志,2011,32(1)：52-53.
[20]刘占军,韩刚.如意金黄散膜剂的改进[J].时珍国医国药,2009,20(7)：1782-1783.
[21]陈永财,周斌,等.正交试验法优选“如意金黄散”巴布剂基质配方[J].海峡药学,2008,20(9)：12-14.
[22]赵浩如,杨永刚,何煜.肉桂超细微粉的粉体特征及体外透皮作用的研究[J].中成药,2002,24(9)：654-656.
[23]严华,魏锋.同属不同种大黄及含大黄制剂中土大黄苷检查方法的研究[J].药物分析杂志,2010,(9)：1615-1620.
[24]张清.黑膏药疮疡膏工艺及质量标准研究[J].时珍国医国药,2008,19(10)：2423-2425.
[25]李明,周大颖,田园,等.中药姜黄的薄层色谱研究[J].贵州化工,2008,(1)：31-34.
[26]王静,秦伟,张全明.TLC法测定散结乳癖贴膏中姜黄素的含量[J].中国药房,2007,18(18)：1394-1395.
[27]张康,何宇新,吴丽等.正交试验优选镇痛贴乙醇提取工艺[J].中国实验方剂学杂志,2011,17(5)：37-39.
[28]杨敏,杨光琼,赵青HPLC法测定如意金黄散中姜黄素的含量[J].现代医药卫生,2011,27(10)：1461-1462.
[29]楼步青.双黄巴布剂体外透皮促进剂的筛选[J].时珍国医国药,2011,22(9)：2247-2250.
[30]吴青青,陈彦.姜黄素脂质体的制备及体外透皮研究[J].中国医药工业杂志,2011,42(12)：910-913.
[31]孙冬梅,李智勇,邓亚利.癌痛巴布剂的质量标准研究[J].江西中医学院学报, 2011,23(4)：34-37.
[32]赵美玉.浅谈巴布剂的生产工艺[J].中国中医药咨讯.2011,3(8)：288-288.
[33]刘方艺,谢友良,蒋东旭.正交设计法优化经络贴巴布剂基质配方研究[J].中国医药导报,2011，8(10)：68-70.
[34]庞来祥,寻园,郭鑫等.正交试验法优化通络止痛巴布剂配方究[J].中国民族民间医药杂志,2011,20(3)：48-49.
[35]徐华明,江国荣.金黄乳膏抗炎镇痛作用的实验研究[J].中医正骨，2011,23(9)：14-16.
[36]刘媛,王健.羊软骨Ⅱ型胶原蛋白对小鼠佐剂性关节炎抑制作用的研究[J].食品科学，2010,(15)：268-270.
[37]陈奇.中药药理研究方法学[M],人民卫生出版社,第1版,1993：378.
[1]陈实功.外科正宗[M].北京：人民卫生出版社,1964：1.
[2]国家药典委员会.中华人民共和国药典[S].北京：化学工业出版社,2010：720-721.
[3]舒林利,李莉.75%乙醇联合如意金黄散外敷治疗静脉炎[J].护理学杂志，2010，25(11)18-19.
[4]邓家英,吴仲安,符关芳.如意金黄散外敷治疗化疗性静脉炎84例[J].中国民康医学,2010,22(1)：11.
[5]林艳容.如意金黄散外敷治疗静脉留置针并发静脉炎[J].浙江中西医结合杂志,2010,20(3)：179.
[6]Bouiton A J. The diabetic foot:aglobal view[J]. Diabetes Metab R es R ev,2000,16 suppll:S2-S5.
[7]贾秀卿,孙喜明.如意金黄散外敷在糖尿病足溃疡护理中的应用[J].临床和实验医学杂志,2010,9(10)：786-787.
[8]和玉英,文歧琳.如意金黄散外敷配合抬高架减轻下肢痛风性关节炎急性期肿痛的疗效观察及护理[J].辽宁中医药大学学报,2009,11(9)：144-145.
[9]王京红.如意金黄散治疗皮肤疮疬肿毒32例[J].中国现代药物应用,2009,3(6)：78.
[10]柴维霞.如意金黄散联合神灯治疗四肢急性软组织挫伤疗效观察[J].中国中医急症,2012,21(1)：170.
[11]王晓,陈欣菊.如意金黄膏封包治疗肝硬化合并自发性腹膜炎58例明[J].中国中医药信息杂志,2007,14(12)：74.
[12]杨玉英,许增宝,朱炜,等.如意金黄散熏洗治疗慢性前列腺炎的效果观察[J].护理与康复,2005,4(2)：86-87.
[13]陈玉涛,王五俊.浓茶冲调如意金黄散外敷辅助治疗阑尾周围脓肿43例临床分析[J].黑龙江中医药,2009,(3)：24-25.
[14]Minghetti, PCasirahgiA, Giluzro, Fetal.Development of local patehes containing melilot extract and ex vivo-in vivo evaluation of skin permeation[J].Euor J Pharm Sci, 2000,10:111-117.
[15]王建明,王政超.抗风湿涂膜剂的制剂研究[J].哈尔滨商业大学学报,2011,27(5)：662-666.
[16]原素敏,倪健.脉炎宁透皮贴剂成型工艺及体外透皮释放研究[J].中国中药杂志,2009,34(13)：1654-1657.
[17]周小凤,杨中林,李萍,等.不同纯度苦杏仁普巴布剂的透皮吸收比较研究[J].药学与临床研究,200,16(4)：285-287.
[18]刘占军,韩刚.如意金黄散膜剂的改进[J].时珍国医国药,2009,20(7)：1782-1783.
[19]陈永财,周斌,等.正交试验法优选“如意金黄散”巴布剂基质配方[J].海峡药学,2008,20(9)：12-14.
[20]刘芳,刘小平.增效如意金黄散软膏制各工艺的研究[J].湖北中医杂志,2006,28(6)：52-53.
[21]韩刚,张卫国,王超,等.高效液相色谱法测定如意金黄散中5种成分的含量[J].中国医院药学杂志,2007,27(1)：124-125.
[22]毛泉明,毛平,叶福嫒,等.反相高效液相色谱法测定如意金黄贴膏中小檗碱的含量[J].中国医院药学杂志,2002,22(8)：477-478.
[23]卢静华,杨欢.双波长薄层扫描法测定如意金黄散中盐酸小檗碱的含量[J].中国生化药物杂志,2011,32(1)：52-53.
[24]江汉美,卢金清,刘璇,等.聚酰胺薄层扫描法测定如意金黄散中厚朴酚与和厚朴酚的含量[J].湖北中医学院学报,2009,11(5)：27-28.
[25]林治琳，林晓瑜.加昧如意金黄散外敷治疗骨折及软组织损伤[J].山东中医杂志,2008,27(10)：681-682.
[26]周龙恒.加味金黄散外敷治疗急性软组织损伤[J].中医正骨,2008，20(10)：68.
[27]兰绍波,宋修亭.如意金黄散外治湿疹132例[J].辽宁中医杂志,2006,33(4)：48.
[28]倪绍军,战广茂.如意金黄散治疗肛周湿疹30例[J].中国民康医学杂志,2005,17(7)：346.
[29]Godin B, Touitou E. Erythromycin ethosomal systems:physi-Cochemical Characterization and enhanced antibacterial activity [J]. Curr Drug Deliv,2005,2(3) 269-275.
[30]Dayan N,Touitou E.Carriers for skin delivery of trihexyphenidyl HCL:ethosomes vs. Liposomes [J]. Biomaterials,2000,21:1879-1885.
[31]Touitou E,Godin B.Dayan N,et al.Intracellular delivery mediated by an ethosomal carrier [J]. Biomaterials,2001,22:3053-3059.
[32]毛平,夏卉莉.如意金黄贴膏与如意金黄散消肿止痛功效的比较研究[J].中国实验方剂学杂志,2004,10(5)：46-48.