【作者】 杨静；

【导师】 韩光；

【作者基本信息】 河南大学 ， 药物分析学， 2015， 硕士

【摘要】 穿心莲有效部位(AEP)中主要的二萜内酯类成分包括穿心莲内酯(A)、新穿心莲内酯(NA)、14-去氧穿心莲内酯(DA)和脱水穿心莲内酯(DDA)等,研究表明,穿心莲二萜内酯类成分具有解热抗炎、抗肿瘤、抗病毒等药理活性,但由于二萜内酯类成分水溶性差、生物利用度较低而影响临床应用。分散片作为片剂的一种新剂型,具有崩解迅速、溶出速度快、生物利用度高等特点。本课题组前期制备了穿心莲有效部位分散片,本论文在此基础上建立穿心莲有效部位分散片的质量标准,并对其进行稳定性研究。本文建立了HPLC法测定穿心莲有效部位分散片中的二萜内酯类成分(A、NA、DA、DDA)的含量测定方法,色谱条件:色谱柱:BDS-HYPERSIL C18柱(250 mm×4.6mm,5μm),柱温:室温,流动相:乙腈-水梯度洗脱,10:90(0 min)→60:40(50 min)→10:90(65 min),检测波长205 nm,流速:1 mL/min,进样量:25μL;理论塔板数以穿心莲内酯计算不低于2000,拖尾因子在0.95-1.10之间,各标准品色谱峰与其相邻峰的分离度均大于1.0;结果显示A、NA、DA、DDA的保留时间分别为25.729 min、33.423min、36.003 min、36.640 min,方法学考察A的回归方程为:Y=2.2×106X-6.0×105,R~2=0.9997,线性范围是2.073μg-10.365μg,NA的回归方程为:Y=2.0×106X-62312,R~2=0.9993,线性范围是0.188μg-0.940μg,DA的回归方程为:Y=2.9×106X-1.9×105,R~2=0.9992,线性范围是0.341μg-1.705μg,DDA的回归方程为:Y=3.7×106X-3.8×105,R~2=0.9994,线性范围是0.581μg-2.905μg;A、NA、DA、DDA的日内精密度RSD值≤1.82%,日间精密度RSD值≤4.71%;样品溶液的重复性RSD值≤3.64%,说明该方法重现性较好,样品溶液在48 h内的稳定性RSD值≤4.18%,回收率试验中A、NA、DA、DDA的RSD值分别是4.37%、2.56%、1.16%、3.62%;穿心莲有效部位分散片中有效成分A、NA、DA、DDA的百分含量分别为11.97%、0.55%、1.72%、2.55%。依照2010版《中国药典》对穿心莲有效部位分散片进行质量评价。采用硬度计测定穿心莲有效部位分散片的硬度,结果表明其承受压力在40 N-60 N范围内;将穿心莲有效部位分散片置温度为20℃±1℃,体积为100 mL水中振摇,记录其崩解时限,结果显示该分散片在3 min内全部崩解并通过了2号筛;对穿心莲有效部位分散片进行片重考察,结果表明该分散片的重量差异限度≤5.0%;另外考察穿心莲有效部位分散片的脆碎度,结果显示其减失重量≤1%;综上,穿心莲有效部位分散片符合药典规定的对制剂的要求。采用理化试验和薄层色谱试验对穿心莲有效部位分散片进行成分鉴别,结果显示穿心莲有效部位分散片的有效成分为二萜内酯类化合物,且阴性对照品试验对其无干扰,专属性强;采用电感耦合等离子原子发射光谱法测定穿心莲药材、穿心莲提取物及穿心莲有效部位分散片中重金属和砷盐的含量,结果如下:穿心莲药材中Pb的含量为4.0114mg/kg,浓度为40.17 ppb,穿心莲提取物中Pb的含量为0.4536 mg/kg,浓度为4.5380 ppb,穿心莲有效部位分散片中Pb的含量为0.2867 mg/kg,浓度为2.8730 ppb;穿心莲药材中As的含量为0.2055 mg/kg,浓度为2.0579 ppb,穿心莲提取物中As的含量为0.0145mg/kg,浓度为0.1451 ppb,穿心莲有效部位分散片中As的含量为0.0158 mg/kg,浓度为0.1583 ppb;对穿心莲有效部位分散片采用浆法测定其在1000 mL 0.3%SDS溶出介质中的溶出度,结果显示其在30 min内的累积溶出度超过95%,且溶出速率较普通片快。依据药物稳定性技术指导原则对穿心莲有效部位分散片进行稳定性研究。影响因素试验,包括高温试验(60℃)、高湿试验(25℃,RH 90%±5%)和强光照试验(4500lx±500 lx),结果表明穿心莲有效部位分散片的性状、鉴别、分散均匀性、水分、重量差异、含量、微生物限度的检查以及溶出度等指标均符合指导原则相关规定;加速稳定性试验,分别于第0、1、2、3、6月末取样,结果表明样品的含量及溶出度等指标均未见明显变化,符合指导原则相关规定;长期试验,分别于第0、3、6、12月末取样,结果表明样品的含量及溶出度等指标均未见明显变化,符合指导原则相关规定。综上所述穿心莲有效部位分散片比较稳定,但需密封保存,其确定的最终有效期为12个月。更多还原

【Abstract】 Diterpene lactones Andrographis(A),neoandrographolide(NA),14-deoxy-andrographolide(DA),dehyd roandrographolide(DDA)and any other ingredients are the effective components that exist in Androg raphis effective parts(AEP).As studies shown that it has the pharmacological activities such as antipyretic,anti-inflammatory,anti-tumor and anti-virus,but clinical application is greatly affected by its poor water solubility property and low bioavailability.As a new dosage form of tablets,dispersible t-ablet characterized by rapid disintegration,dissolution rate and high bioavailability.Our team prepare-d the Andrographis effective parts dispersible tablets at the early stage and established its quality standards on the basis of former research and further studied its stability.A method for testing the contents of A、NA、DA、DDA in Andrographis effective dispersible tablets by HPLC are described in this paper.The parameters setup of chromatographic conditions are as follows:chromatographic column:BDS-HYPERSIL,C18 column(250 mm×4.6 mm,5 μm);Column temperature:room temperature;mobile phase: acetonitrile-water gradient,10:90(0 min)→60:40(50 min)→10:90(65min);detection wavelength:205 nm;flow rate: 1mL/min;injection volume:25 μL;theoretical plate numbers were not less than 2000 calculated by andrographolide;trailing factor was between 0.95 and 1.10;the separable degree should above 1.0 between chromatographic peak of standard and adjacent peak.The results showed that A,NA,DA,DDA retention time was 25.729 min,33.423 min,36.003 min,36.640 min.Meanwhile,the regression equations of these four components by methodology are as follows:A:Y=2.2×106X-6.0×105,R~2=0.9997,linear range:2.073 μg-10.365μgNA:Y=2.0×106X-62312,R~2=0.9993,linear range :0.188μg-0.940μgDA :Y=2.9×106X-1.9×10~5,R~2=0.9992,linear range :0.341μg-1.705μgDDA :Y=3.7×106X-3.8×10~5,R~2=0.9994,linear range : 0.581μg-2.905μg.At the same time,the day precision RSD value and inter-day precision RSD value of A,NA,DA,DDA are separate less than 1.82% and 4.71% along with the RSD value of sample solution repeatability are less than 3.64% which all showing that this method is good.The RSD value of sample solution stabilitywithin 48 h is smaller or equal to 4.18% and recovery test in A,NA,DA,DDA,RSD values are 4.37%,2.56%,1.16%,3.62%.Besides,the percentage of effective components of A,NA,DA,DDA in Andrographis effective dispersible tablets are 11.97%,0.55%,1.72%,2.55 %.The quality evaluation of Andrographis effective parts dispersible tablets was carried on according to the《China Pharmacopoeia》(2010 edition).The hardness tester was employed to determine the hardness of Andrographolide effective parts dispersible tablet and the results showed that the pressure is between 40 N and 60 N.Andrographis effective parts dispersible tablets disintegrate within 3min and pass through the 2nd sieve when putting in water of 20℃±1℃ and 100 mL.Meanwhile,the difference between tablet weight is smaller than or equal to 5.0% and weight loss is smaller than or equal to 1% when tested its tablet weight and frangibility which all showing that the Andrographis effective parts tablets comply with regulations of preparation according to the pharmacopoeia.The Andrographis effective parts dispersible tablets are diterpenoid compounds when tested by the method of physical and chemical tests and thin-layer chromatograph.It also showed that control test confirmed its non-interference which indicated its strong specificity.Simultaneously,the heavy metal and arsenic salt in tablets were evaluated by IOES and the results are as follows:Pb content in Andrographis paniculata is 4.0114 mg /kg,concentration is 40.17 ppb;Pb content in Andrographis extract is 0.4536 mg/kg,concentration is 4.5380 ppb;Pb content in Andrographis effective parts dispersible is 0.2867mg/kg,concentration is 2.8730 ppb;As content in Andrographis paniculata is 0.2055 mg/kg,concentration is2.0579 ppb;As content in Andrographis extract is 0.0145 mg/kg,concentration is 0.1451 ppb;As content in Andrographis effective parts is 0.0158 mg/kg,concentration is 0.1583 ppb.We used grouting method to determine the in vitro release profile with 1000 mL 0.3% SDS as the release medium,and the results showed its cumulative release percentage in 30 minutes is more than 95% and the dissolution rate is faster than ordinary tablets.Based on technical guidelines for drug stability,the stablity of Andrographis effective parts tablets was investigated.The impact factors included high temperature test(60℃),high humidity test(25℃,RH90 %±5%)and strong lighting pilot(4500 lx±500lx).The results showed that the trait,identification,dispersion uniformity,moisture,weight variation,content,microbial limit,dissolution of the inspection and other indicators of Andrographis effective part tablet are all standards-compliantin.Nosignificant changes are found in content and dissolution both in accelerated stability test and long-term test which were respectively investigated at the end of 0,Jan,Feb,Mar,Jun and 0,Mar,Jun,Dec.They are all in line with the guiding principles.In summary,Andrographis effective parts tablets are relatively stable,but need be sealed for storage and the ultimate validity are 12 months.更多还原