蒙药清血八味片质量控制方法研究

发布时间：2018-08-20 11:40

【摘要】：目的蒙药清血八味片由寒水石(凉制)、紫草、土木香、人工牛黄、栀子、瞿麦、石膏、甘草等八味药材组成,为蒙医临床常用验方,在治疗血病方面疗效显著。清血八味片的物质基础研究薄弱,现有质量标准很不完善,亟需提高质量控制水平。本课题拟建立清血八味片的指纹图谱,对清血八味片中多指标性成分进行含量测定方法研究,定性定量分析清血八味片所含无机元素,实现对清血八味片多方面的质量控制方法研究,从而更加全面的控制产品质量。方法1.利用HPLC-PDA技术建立清血八味片的指纹图谱,将18批清血八味片样品色谱图采用相似度软件进行处理,对建立的指纹图谱及其各项技术参数进行分析。利用各单味药材谱图对清血八味片中共有峰进行归属,通过已有对照品进行部分特征峰的指认。2.以硝酸加过氧化氢体系为消解试剂,供试品经微波消解后,采用电感耦合等离子体质谱(ICP-MS)法测定清血八味片中 Be、B、Na、Mg、Al、P、K、Ca、Ti、V、Cr、Mn、Fe、Co、Ni、Cu、Zn、Ga、As、Se、Rb、Sr、Mo、Ag、Cd、Sn、Sb、Cs、Ba、Hg、Tl、Pb共32种无机元素的含量,绘制无机元素含量分布曲线建立清血八味片无机元素指纹谱,用SPSS 19.0软件对数据进行主成分分析并筛选主要特征元素。3.由于寒水石及石膏主成分相同,故对两者所含钙总量进行测定。通过理化鉴别试验进行清血八味片的钙盐与硫酸盐的鉴别反应;对清血八味片的样品处理条件进行合理优化,并对测定方法进行考察;将样品经炭化后灰化,采用乙二胺四乙酸二钠(EDTA-2Na)络合滴定法对清血八味片中钙成分进行含量测定。4.选择清血八味片中主要活性成分京尼平龙胆二糖苷、栀子苷、甘草苷、甘草酸、异土木香内酯和土木香内酯作为指标性成分,通过单因素考察选择最佳供试品提取方法,并对色谱检测条件进行优化,利用高效液相色谱法同时测定18批样品中6个指标性成分的含量。结果1.清血八味片HPLC色谱图中各成分得到了良好分离,确定了 21个共有指纹峰,初步阐明了共有峰来源并定性了部分色谱峰。将18批次样品色谱图与生成的对照指纹图谱进行匹配,平均相似度为0.974,RSD为1.54%,表明18批样品生成的图谱的相似度较高,差异较小。所建立的指纹图谱方法可行性较好。2.ICP-MS法测定的32种无机元素线性关系良好,相关系数r ≥ 0.9995,各元素的检测限在 0.001～6.390 μg/L,加样回收率为 94.36%～105.47%,RSD 为 1.53%～4.56%。通过无机元素含量分布曲线分析建立了清血八味片的无机元素指纹谱,不同批次的样品均有相似的峰形,且不同无机元素含量的多寡顺序趋于一致。经主成分分析筛选出清血八味片主要特征无机元素为 Co、As、Ba、Ca、Sb、Ti、Ni、Fe、Cs、Se、P、Al、K。测定的样品中检测出Pb、Cd、As、Hg、Cu,5种重金属元素的含量均低于《中国药典》2015版的限度要求。3.采用络合滴定法测定清血八味片中钙含量,样品最佳处理条件是将样品在调压电炉上炭化1 h至炭化完全后,置于马弗炉中在700～800℃下灰化6 h。测定方法的重复性试验RSD为0.32%,加样回收率为98.67%～101.25%,RSD为0.97%,均满足定量分析的要求。4.采用HPLC法测定清血八味片中京尼平龙胆二糖苷、栀子苷、甘草苷、甘草酸、异土木香内酯和土木香内酯等被测组分均达到基线分离,在一定浓度范围内6种成分的线性关系良好,相关系数r≥0.9998,精密度、重复性、稳定性、加样回收率等内容均符合定量分析要求。18批样品的6种成分含量较稳定,无显著性差异。结论本课题通过对蒙药清血八味片进行指纹图谱、无机元素分析、多成分含量测定的研究,为该产品质量标准的提升和产品质量的全面控制提供技术参考,为该蒙成药的生产加工、市场流通、药效物质基础研究、临床应用等提供科学依据。
[Abstract]:OBJECTIVE Qingxue Bawei tablets are composed of eight kinds of medicinal materials, such as cold water stone, Shikong grass, earthwood fragrance, artificial bezoar, Gardenia jasminoides, Trichosanthes, gypsum and liquorice. It is a common prescription in Mongolian medicine and has remarkable curative effect in treating blood diseases. To establish the fingerprint of Qingxue Bawei Tablets, to study the content determination method of multi-index components in Qingxue Bawei Tablets, to analyze the inorganic elements in Qingxue Bawei Tablets qualitatively and quantitatively, and to study the quality control method of Qingxue Bawei Tablets in many aspects, so as to control the quality of products more comprehensively. Methods 1. Using HPLC-PDA technology. The fingerprint of Qingxuebaobawei tablets was established. The chromatograms of 18 batches of Qingxuebaobawei tablets were processed by similarity software. The fingerprint and its technical parameters were analyzed. Nitric acid plus hydrogen peroxide system was used as digestion reagent. After microwave digestion, the contents of Be, B, Na, Mg, Al, P, K, Ca, Ti, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Ga, As, Se, Rb, Sr, Mo, Ag, Cd, Sn, Sb, Cs, Ba, Hg, Tl and Pb in Qingxue Bawei tablets were determined by inductively coupled plasma mass spectrometry (ICP-MS). To establish the fingerprint spectrum of inorganic elements in Qingxue Bawei Tablets, principal component analysis and screening of main characteristic elements were carried out with SPSS 19.0 software. After carbonization and ashing, the content of calcium in Qingxue Bawei tablets was determined by EDTA-2Na complexometric titration. 4. Geniposide, geniposide, glycyrrhizin were selected as the main active components in Qingxue Bawei tablets. Glycyrrhizic acid, iso-tufa lactone and tufa lactone were used as index components, and the best extraction method was selected by single factor investigation. The chromatographic detection conditions were optimized. The content of 6 index components in 18 batches of samples was determined by high performance liquid chromatography. Results 1. The components in the HPLC chromatogram of Qingxuebaobawei tablets were obtained. Twenty-one common fingerprint peaks were identified by good separation, and the source of common peaks was elucidated preliminarily. The fingerprint of 18 batches of samples were matched with the corresponding fingerprints with an average similarity of 0.974 and RSD of 1.54%. The results showed that the similarity of the fingerprints produced by 18 batches of samples was high and the difference was small. The correlation coefficient R (> 0.9995), the detection limit of each element is 0.001-6.390 ug/L, the recovery is 94.36%-105.47%, RSD is 1.53%-4.56%. The fingerprint spectrum of inorganic elements in Qingxuebaobawei tablets is established by analyzing the content distribution curve of inorganic elements. The main characteristic inorganic elements of Qingxuebai Bawei tablets were Co, As, Ba, Ca, Sb, Ti, Ni, Fe, Cs, Se, P, Al, K. The contents of Pb, Cd, As, Hg, Cu and 5 heavy metal elements were lower than those in the Chinese Pharmacopoeia > 2015. Calcium content in Qingxue Bawei tablets was determined by complexometric titration. The optimum conditions for sample treatment were carbonization for 1 h in a pressure regulating furnace and ashing for 6 h in a muffle furnace at 700-800 C. The repeatability test RSD of the method was 0.32%. The recovery of sample addition was 98.67%-101.25% and RSD was 0.97%. Quantitative analysis of Geniposide, Geniposide, Geniposide, Glycyrrhizin, Glycyrrhizic acid, Isoamylactone and Cinnamolide in Qingxuebai Bawei Tablets were determined by HPLC. The six components were separated at baseline. The linear relationship of the six components was good within a certain concentration range. The correlation coefficient R (>0.9998), precision, repeatability and stability were obtained. The contents of six components in 18 batches of samples were stable, and there was no significant difference between them.Conclusion The research on fingerprint, inorganic element analysis and multi-component content determination of Mongolian medicine Qingxue Bawei tablets can provide a reference for the improvement of product quality standard and the overall control of product quality. Technical reference will provide scientific basis for the production, processing, market circulation, basic research of pharmacodynamic substances and clinical application of the Mongolian patent medicine.
【学位授予单位】：北京中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R29

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