【作者】 柳俊；

【导师】 涂瑶生；

【作者基本信息】 广州中医药大学 ， 中药学， 2011， 博士

【摘要】 目前,有关中药配方颗粒生产过程的质量控制研究一般是针对少数关键生产环节中间体及工艺终点的产品进行质量分析,而关于其生产过程在线质量监控的研究鲜有报道。中药配方颗粒生产过程为典型的批次生产过程,其生产过程由一系列单元操作如提取、浓缩、干燥等组成,每一项单元操作工艺参数的变化都会影响到最终产品的质量。由于缺乏有效的在线质量分析方法,生产过程往往只能依赖经验进行操作,根据经验决定过程是否完成,从而导致产品难以达到国际上对药品质量稳定均一的要求,严重制约了中药配方颗粒工业化、国际化的发展。近红外光谱技术(Near-Infrared Spectroscopy,NIRS)是目前最常用的过程分析技术(Process Analytical Technology,PAT)之一,它是20世纪80年代后期迅速发展起来的一种新型分析检测方法,它不需对样品进行复杂的预处理就可直接进行测试,测试过程通常在几分钟之内就能完成,而且整个测试过程不需要使用和消耗任何化学试剂,是一种“绿色、环保”型分析检测方法,已在农产品、石油化工、生命科学及药物分析等多个领域得到广泛应用。由于中药配方颗粒为单味药材提取物,与复方中药相比,其物质组成简单,基础研究较深入,是NIRS技术在中药生产质量控制示范研究的良好载体。将近红外光谱分析技术应用于中药配方颗粒制备过程质量控制,建立过程中关键操作单元如提取、浓缩、干燥等近红外(NIR)在线监测方法,及时从实际制备过程中得到物料性质信息,了解关键过程参数与最终产品质量之间的关系,对制备过程工艺参数进行即时调控,改变目前根据经验决定过程是否完成的质控手段,从而达到控制和优化药物制备过程的目的,使得不同批次的产品达到质量稳定均一的要求,促进中药配方颗粒进一步的发展。本文以白芍配方颗粒制备过程为具体研究对象,开展了近红外光谱分析技术在白芍配方颗粒制备过程重要生产环节中的应用研究及Box- Behnken效应面法在优化中药提取工艺的应用研究,取得了较满意的结果。1.中药药材的质量控制在整个中药配方颗粒生产过程中占有重要的地位,不同产地及不同批次的药材质量的稳定与否直接影响到后续工序的操作和产品的最终质量。本文所选用的白芍药材均来自全国各不同产地,具有一定的代表性,所有样品均在同一条件下进行粉碎,并过80目药典筛。运用偏最小二乘法(PLS)建立白芍药材粉末近红外光谱(NIR)与其芍药苷、芍药内酯苷和水分含量测定值之间的多元校正模型,校正模型相关系数(R2)分别为0.938、0.943、0.976。对验证集样品进行预测,预测结果表明,该法可以较准确地预测样品中的芍药内酯苷和水分的含量,预测平均相对偏差分别为0.23%、3.82%,而部分样本芍药苷的含量偏低,在近红外光谱法的检测低限,测定效果不甚理想。近红外光谱法具有分析速度快、预测结果准确、不破坏样品和不污染环境等优点,而且不需要对样品进行复杂繁琐的前处理,适合对组成复杂的中药进行快速分析,可用于白芍配方颗粒原料药材的质量监控。2.Box-Behnken效应面法是采用多元二次方程拟合因素和效应值之间的函数关系,并通过对回归方程的分析寻求最优工艺参数,解决多变量问题的一种统计方法。本文以白芍总苷提取量为量化指标,在单因素实验的基础上,探索了用Box-Behnken效应面法确定白芍总苷提取的最优工艺参数的可行性。取得了比较满意的结果,并最终确定其最佳提取工艺参数为：浸泡时间为98.4min,加水量为12倍,提取时间为69.8min。在此条件下,白芍总苷的提取量为2.39%,与理论值仅相差0.014%,从而说明了Box- Behnken效应面法在中药提取工艺应用中的可行性。3.中药提取过程是中药配方颗粒生产过程中的关键环节,缺乏主要成分含量在线监控是制约产品质量稳定性的瓶颈。本文将白芍一次提取和二次提取过程作为研究对象,运用近红外光谱法结合PLS法建立白芍提取液中芍药内酯苷及芍药苷含量的定量校正模型,结果表明所建芍药内酯苷和芍药苷含量NIR定量校正模型的预测能力较好,样品中芍药内酯苷和芍药苷含量预测值与参考值之间的偏差较小,预测值的平均相对偏差分别为1.51%,-1.87%,预测平均回收率分别为100.48%,98.13%,说明所建立的近红外光谱定量校正模型可以用于实时监控白芍提取过程中提取液所含芍药内酯苷、芍药苷含量的变化,并能对提取过程终点进行判断。4.浓缩是中药配方颗粒生产过程中的重要操作单元,不同批次浸膏质量的稳定与否直接影响到后续工序的操作和产品的最终质量。本文以白芍水提液浓缩过程为例,将近红外光谱技术用于白芍水提液浓缩过程在线分析。利用PLS2法建立的近红外光谱校正模型能够快速预测浓缩液中芍药内酯苷和芍药苷的含量,及时获得浓缩过程药液有效成分的信息,并对浓缩终点作出判断。模型预测值与参考值之间具有较好的相关性,预测相对平均偏差分别为-1.16%,0.94%,表明利用近红外光谱技术能实时反映浓缩过程中料液的状态,提高中药浸膏质量的稳定性。5.喷雾干燥法具有干燥速度快、受热时间短、生产工序少、产品流动性好、质地均匀、溶解性好等特点。近年来,我国中药产业发展迅速,在众多的中药制药企业中,喷雾干燥技术应用极其广泛。本文在单因素实验的基础上,以产物水分含量为指标,采用正交试验法研究喷雾干燥时料液相对密度、进/出风温度、进料速度对白芍提取物喷雾干燥的影响,并确定了最佳喷雾干燥工艺参数为：进/出风温度为190/90℃,料液相对密度1.08(60℃),进料速度为7mL/min。6.本文将白芍提取液喷雾干燥过程作为研究对象,综合考察提取操作参数对产物水分含量及芍药内酯苷、芍药苷含量的影响,运用NIR光谱法结合PLS法和PCR法建立白芍喷干粉中水分、芍药内酯苷以及芍药苷含量的定量校正模型,结果表明上述所建水分、芍药内酯苷和芍药苷含量NIR定量校正模型的预测能力较好,样品中水分、芍药内酯苷和芍药苷含量预测值与参考值之间的偏差较小,预测值的平均相对偏差分别为-1.38%,0.30%、0.12%,预测平均回收率分别为98.62%,100.30%、100.12%,说明近红外光谱技术适用于喷雾干燥过程的在线质量控制。以上研究结果表明,近红外光谱分析技术可用于白芍配方颗粒制备过程主要环节的过程质量控制。如能结合本文研究成果,并根据中药配方颗粒的生产特点,建立一套完善的近红外在线检测体系,实时监控中药配方颗粒生产加工过程,通过分析原料药及其制品加工过程中物理性质及化学成分指标变化来控制中药配方颗粒的质量状况,并且建立完善的近红外检测中药配方颗粒的模型库并不断扩充模型,这样就可以有效的控制中药配方颗粒的质量,确保产品的质量稳定和可重复性。相信近红外光谱技术在中药配方颗粒生产领域将具有非常广阔的应用和发展前景。更多还原

【Abstract】 At present, the quality control of the Traditional Chinese Medicine (TCM) formula granule production process generally involves only the content ana-lysis at the end of the process, and on-line quality control of the process has been reported rarely. Production process of TCM formula granule is typical batch process, consist of a series units such as extraction, concentration, drying, etc., a change in parameter of each unit will affect the final product quality. Lack of effective on-line quality analysis method, production oper-ations often rely on experience, decision process is complete based on exp-erience, leading to product hard to reach for international drug quality stablility requirements, which seriously restricted the TCM formula granule industrialization and internationalization development.Near infrared spectroscopy (NIRS) is the most commonly used technology for process analysis, which is a new type of detection method, developing rapidly since the late 1980s, the samples does not need complex pretreatment can be directly tested, the testing process is usually done in a few minutes, and the whole testing process doesn’t need to use any chemical reagent, it is a kind of environmental protection analysis and detection methods, it has been widely used in agricultural, petrochemical, life sciences and pharmaceutical analysis and other fields.Near infrared spectroscopy technology applied in TCM formula granule production process, establishing near infrared (NIR) on-line monitoring method in the production process key operation units such as extraction, concentra-tion, drying, etc., getting the material properties information from the actual production process timely, to understand the relationship between the key process parameters and the quality of the final product, thus to achieve production process parameter real-time control and optimization, making different batches of product achieve quality stablility requirements, and promoting the further development of TCM formula granule.In this paper, the application of NIRS technique to on-line monitor the production process of peony formula granule had been researched, and achieved satisfactory results.1. Quality control of herbal medicine in the whole production process of TCM formula granule plays an important role, the quality of medicinal materials which come from different areas and different batches exist certain differen-ces, which directly affects the subsequent operation process and the final product quality. Radix Paeoniae Alba selected in this paper come from different areas across the country, has certain representativeness, all samples were crushed under the same conditions, and pass 80 mesh pharmacopoeia sieve. Multivariate calibration models based on partial least squares (PLS) algorithm were developed to correlate the spectra and the corresponding values determ-ined by the reference method. The corelation coefficients (R2)of the calibra-tion models were 0.938、0.943 and 0.976, and the prediction average relative deviation for paeoniflori、albiflorin and moisture content were 6.46%、0.23% and 3.82%, respectively. The NIRS determination method was rapidly, accuracy, nondestructive and non-pollution. It can dispose the samples without compli-cated pretreatment. It is qualified to analyze TCM whose components are complex rapidly. NIRS can be used to control the quality of herbal material of paeony formula granule.2. Box-Behnken response surface method is a statistical method that use multiple quadratic equation to fit the relations between factors and the responses, through the analysis of the regression equation to find the optimum process parameters,to solve the multivariate problems. In this article, base on the results of single factor experiment, Box-Behnken experimental design combining with response surface methodology(RSM) was employed, with the soa-king time (X1), the water addition (X2), the extraction time (X3) as the inde-pendent variables. The response variable was the amount of Total Glucoside of Paeony. The optimization Processing Parameters was as follows:X1，X2and X3 levels of 98.4min,12 times and 69.8min, respectively. The observed responses are in close agreement with the predicted values of the mathematic models, and the Box-Behnken response surface method is suitable for optimizing the extraction of TCM.3. The extraction process of TCM is a key step in the production process, due to lack of on-line monitoring method, it is difficult to ensure the sta-bility of product quality. In this paper, the extraction process as the research object, multivariate calibration models based on partial least squares (PLS) algorithm were developed to correlate the spectra and the corresponding values determined by the reference method. The prediction average relative deviation for the content of albiflorin、paeoniflorin were 1.51%、-1.87%, respectively, the prediction average recovery were 100.48%,98.13%, respectively. That the established quantitative NIR calibration model can be used in the extraction process to on-line monitor the content changge of albiflorin and paeoniflorin.4. Extraction is an important step in the TCM production process, the quality of different batches extract exist differences, which directly affects the subsequent operation process and the final product quality. In this paper, the concentration process as the research object, multivariate calibration models based on PLS2 algorithm were developed to correlate the spectra and the corresponding values determined by the reference method. The prediction average relative deviation for the content of albiflorin、paeoniflorin were-1.16%、0.94%, respectively. That the use of near infrared spectroscopy reflect the enrichment process in real-time liquid state, to improve the quality of traditional Chinese medicine extract stability. Through using the NIRS technology, the situation of medicine liquid in the extraction process can be reflected, Therefore, the stability of the extract quality to obtain improved.5. The spray drying method has the characteristics such as fast-drying, heating time is short, less production processes, good mobility, homogeneous texture, good solubility,etc. In recent years, TCM industry has developed rapidly, the spray drying technology is widely used in many pharmaceutical enterprise. In this paper, the optimum technological conditions of the spray drying process of paeony extract were researched. According to the single factor experiment and the orthogonal test of 3 factors and 3 levels was ado-pted, and the optimum combination was obtained. the result showed the optimum conditions of spray drying process were as followed:inlet air temperature and outlet air temperature were 190℃and 90℃, the relative density of the extract was 1.08 (60℃), inlet flow rate was 7mL/min. 6.In this paper,NIRS technology applied in the spray drying process of paeony extract were researched, multivariate calibration models based on PLS and PCR algorithm were developed to correlate the spectra and the corresponding values determined by the reference method. The prediction average relative deviation for the content of moisture content、albiflorin、paeoniflorin were-1.38%、0.30%,0.12%, respectively, the prediction average recovery were 98.62%, 100.30%,100.12%, respectively. That the established quantitative NIR calibration model can be used in the spray drying process.The research results show that NIRS technology can be used to on-line monitor the major parts of production process of Paeony formula granule. If this research achievement and the characteristics of TCM formula granule production process are combined, set up a complete NIRS on-line detection system to monitor the production process, and improve the detection model of NIRS continually, This can control the quality of TCM formula granule effectively and ensure the stablility and repeatability of product quality.更多还原