基于系统量化指标的阳春砂商品规格及其行业标准的研究
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摘要
中药材商品规格是药材品质的体现,是评价中药材质量好坏最直观的方法。1984年我国颁布实施了第一部关于中药材商品规格的标准《76种药材商品规格标准》,该标准按照质量的高低、好次对七十六种常用中药材进行了等级的划分,规定了品别、规格、等级三个方面的标准。但随着我国中药材商品生产、流通的不断变化和和质量标准的不断更新,原有的标准已不能符合当前中药材的生产、流通和使用。目前中药材市场标准随意性大,部分药材商品规格、等级混乱,没有继承中药材商品规格传统的鉴别经验,也缺乏科学性、系统性的技术资料支持,商品规格标准不能在中药材质量标准评价体系中发挥应有的作用。因此中药材商品规格评价标准的技术方法有必要进一步提高和规范,中药材商品规格等级亟待量化标准的建立。本研究提出中药材基于系统量化指标的商品规格综合评价方法,用于评价和制定中药材砂仁商品规格的行业标准。
砂仁来源于姜科植物阳春砂Amomum villosum Lour.、绿壳砂Amomum villosum Lour. var. xanthioides T. L. Wu et Senjen,或海南砂Amomum longiligulare T. L. Wu的干燥成熟果实,是我国著名“四大南药”之一。目前我国市场流通的砂仁主要以阳春砂为主,都是以“统货”进行收购、加工和销售,无规范的商品规格及其行业标准。本研究采用文献整理、现场调研等溯源式研究,分析阳春砂药材商品规格的形成模式,通过理化实验结合道地性形成模式研究,构建“基原鉴定—性状鉴定—理化鉴定—生物检定—常规检测”为主要内容的阳春砂商品规格行业标准草案,为阳春砂商品规格等级划分的客观化、规范化、标准化提供科学依据,为提高和规范中药材商品规格等级评价标准提供技术方法参考。
一、研究内容和主要研究结果
(一)阳春砂文献调研、道地药材产区的现场调研与取样
1、通过文献调研及对阳春砂主产区的现场调研,以及中药材专业市场、药材公司等的调研和咨询,发现目前市场上“阳春砂”均以“统货”进行收购、销售,不分等级,尚未有不同等级阳春砂的科学分类标准和行业质量标准。
2、文献及主产区调研结果表明,道地药材阳春砂生长需要特定的自然生态环境条件,特别与种植地的湿度、温度、日照、土壤等生态因子密切有关。由于栽培地生态条件的不断变化、栽培技术上的差异以及种质自身的变异,在产区也形成了一些栽培品种(如广东阳春市的“长果”、“圆果”和选育中品种“春选”,云南西双版纳的“高杆”、“中杆”、“低杆”等)。
3、虽然广东省阳春市制订了阳春砂的分级(农业)标准,但当地在收购、销售阳春砂时,并未按照分级标准进行,均以“统货”进行收购、销售。此外,由于市场长期认为广东省阳春市产的阳春砂质量好,因此阳春产的阳春砂药材价格高(300-500元/千克),收购外地产阳春砂加工成干果作为阳春产阳春砂销售的情况存在,也导致在药材市场上有些商家将“非阳春产阳春砂”当作是“阳春产阳春砂”药材进行销售。药材公司通常会挑选个大、饱满度较好的阳春砂供应出口。
(二)阳春砂不同商品规格等级定量化指标的研究
借鉴农产品等商品的感观评价思路和方法,采用特尔菲(Delphi)法筛选出阳春砂商品感官评价的“重量、横径、纵径”三个属性指标；设计阳春砂药材商品感官特征项目,测量和收集与阳春砂商品感官判断相关的指标,结合计算机数据库技术,研究开发了“阳春砂药材商品数据管理系统”。进行感官属性相关度研究,对专家等级评判结果与相应阳春砂样品的果实横径、纵径、重量三个属性间的数值关系进行数据分析实验,得出专家等级评分与果实横径、重量相关性最高。使用稳健回归方法产生预测模型,加上参数拟合线性拟合方程得出了阳春砂不同等级的计算公式,得出了一、二、三等品阳春砂的商品规格等级性状差异,得出样品等级计算的线性方程。研究建立了一、二、三等品阳春砂的商品规格等级。
阳春砂商品专属性状：干货。呈椭圆形或卵圆形,有不明显的三棱。表面棕褐色,密生刺状突起,顶端有花被残基。果皮薄而软。种子集结成团,具三钝棱,中有白色隔膜,将种子团分成3瓣。种子为不规则多面体,表面棕红色或暗褐色,有细皱纹,外被淡棕色膜质假种皮；质硬,胚乳灰白色。气芳香而浓烈,味辛凉、微苦。无杂质、虫蛀、霉变。
一等品阳春砂：果实重量1克以上,果实纵径17毫米以上,果实横径14毫米以上。种子团每瓣有种子16-24粒。种子直径2-3毫米。
二等品阳春砂：果实重量在0.55-1克之间,果实纵径在14-17毫米,果实横径在12-14毫米之间。种子团每瓣有种子12-15粒。种子直径2-3毫米。
三等品阳春砂：果实重量0.55克以下,果实纵径14毫米以下,果实横径12毫米以下。种子团每瓣有种子7-11粒。种子直径2-3毫米。
(三)基于化学分析和生物检测的阳春砂不同商品规格比较研究
基于《中国药典》标准的药效指标成分含量测定、阳春砂药材代谢产物、农残、重金属等检测、化学成分含量高低与商品规格等级的相关性、化学成分含量比例与商品规格等级的相关性,以及基于生物检测的阳春砂药材商品规格比较研究结果如下。
1、各等级商品阳春砂挥发油含量均符合《中国药典》(2010年版一部)规定,不同等级间的阳春砂挥发油含量差异不大。
2、各等级商品阳春砂乙酸龙脑酯含量均符合《中国药典》(2010年版一部)规定,乙酸龙脑酯的含量是一等品>二等品>三等品。采用“层次聚类分析方法”中的欧式距离平方对三个等级阳春砂的乙酸龙脑酯进行聚类分析,二等品和三等品聚为一类,一等品单独为一类,即二等品和三等品的乙酸龙脑酯含量无明显差异,一等品显著高于其他两个品级。
3、各等级商品阳春砂水分含量、有害重金属、农药残留量均符合《中国药典》(2010年版一部)规定。
4、阳春砂挥发油的气质联用色谱分析结果表明,阳春砂挥发油主要峰相对百分含量排序依次为乙酸龙脑酯、樟脑、d-柠檬烯、冰片、莰烯等共十七种挥发性化合物,其中乙酸龙脑酯为阳春砂挥发油中的主要成分,占挥发油总量的45%以上。从系统聚类结果分析,各等级阳春砂挥发油化学组成与相对含量无明显的差异,但挥发油中各成分平均百分比含量结果为一等品>二等品>三等品。
5、以冰乙酸致小鼠扭体实验为生物效价指标的研究结果表明,各等级阳春砂挥发油均有镇痛作用,其中一等品阳春砂镇痛的量效、时效关系比较明确,14%浓度一等品阳春砂挥发油以20mL/kg灌胃,从10min至160min均有镇痛作用,提示一等品镇痛起效较快,维持时间较长。二等品、三等品阳春砂挥发油的镇痛药效相对不稳定,14%浓度二等品阳春砂挥发油在给药80min时对冰乙酸致小鼠扭体有镇痛作用,14%浓度三等品阳春砂挥发油在给药80min和160min时有镇痛作用,两者无差异。
(四)阳春砂药材商品规格等级标准(草案)的建立
参照现版《中国药典》和研究结果,起草阳春砂药材商品规格等级标准(草案),主要包括基原、加工方法、性状、商品规格等级、鉴别、检查、含量测定、生物效价检测、贮藏等方面,形成“基原鉴定—性状鉴定—理化鉴定—生物检定—常规检测”为主要内容的阳春砂商品规格行业标准草案。
二、研究结论
本论文通过参考和借鉴农产品等商品规格评价的方法,结合计算机数据库技术,对实验数据进行分析建模实现对阳春砂商品规格的分级预测,并进一步通过“商品规格—化学成分—生物效应”的关联分析,探讨了阳春砂药材商品规格等级的科学合理性。研究建立了基于系统量化指标的阳春砂药材商品规格等级标准,研究方法可为果实类中药材的商品规格等级研究提供参考和借鉴。
本研究丰富了阳春砂“地理标志产品”商品规格的技术内涵,可为阳春砂道地产区阳春砂GAP过程中的采收、加工、贮藏和商品销售提供科学指导,为阳春砂优质优价提供依据,为进一步制订阳春砂行业标准和管理政策提供依据。
Product specification is a qualitative reflection of Traditional Chinese Medicine (TCM). It's the most intuitive indicator to evaluate TCM."Product Specification Standard of76Drugs", the first norm for product specification of TCM promulgated and implemented in China1984, set classification and standard on product type, specification and grade of76common TCM, according to those drugs' quality. However, with TCM's continuous shifting of product manufacturing and circulation, as well as their renewing of quality standard, this norm does not correspond with the current manufacturing, circulation and apply of TCM. In the current TCM market, no unitive quality standard is acknowledged by the druggists. Because of the complicated product specification and grade of some drugs, the failure of inheriting traditional discerning experiences, and the absence of scientific and systematic technical information, product specification standard cannot play its role in the quality standard evaluating system of TCM's product specification. Therefore, it's necessary to regulate and improve the technology in TCM's product specification evaluating system, and to set a qualitative standard for TCM's product specification and grade. In this study, an integrated evaluating method is established to form and evaluate the industrial standard for the drug Fructus Amomi, based on systematic quality index.
One of China's well-known "Four Southern Medicine", Fructus Amomi, comes from dry mature fruit of Zingiberaceae plants, including Amomum villosum Lour., Amomum villosum Lour. var. xanthioides T. L. Wu et Senjen and Amomum longiligulare T. L. Wu. The main circulation specie in the current TCM market is Amomum villosum Lour.. It's purchased, processed and distributed as unitive goods, without regulating product specification and industrial standard. In this study, traceable research methods, such as literature reviewing and site investigating, were applied to analyze the formation model of Fructus Amomi's product specification. By physical and chemical experiments, together with authentic formation model study, a draft industrial standard for Fructus Amomi's product specification was established to provide scientific basis for objective, norm and standardized classification of Fructus Amomi's product specification and grade. This standard was based on the "original identification-character identification-physical and chemical identification-bioassay-normal detection". Therefore, this study provides technic reference to regulate and improve evaluating standard for TCM's product specification and grade.
1Research contents and main findings
1.1Literature review, site investigation and sample collection of Fructus Amomi
1.1.1Through literature reviewing, site investigating and investigating on TCM market and enterprises, we found that Fructus Amomi was purchased, processed and distributed as unitive goods, without a scientific classifying standard and industrial standard for different grade of the product.
1.1.2Results of literature reviewing and site investigating showed that it needs special nature environmental conditions, especially ecological factors, such as humidity, temperature, sunlight and earth, for Fructus Amomi to grow well. Due to the continuous variation of ecological conditions and cultivable technology, as well as the mutation of the specie itself, new cultivable species could be formed in the generating districts.("Long fruit","round fruit","spring selected" species in Yangchun City, Guangdong province, and "high trunk","medium trunk" and "short trunk" in Xishuangbanna, Yunnan province.)
1.1.3Although classification standard for Fructus Amomi had been made by Yangchun City, Guangdong province, Fructus Amomi were purchased and processed as unitive goods, without classifying according to the standard. Besides, because Fructus Amomi produced by Yangchun City was consumed the best in the TCM market, its price(300-500￥/kg) is higher compared to other species. Illegal phenomenon do exist that some druggists put Fructus Amomi not produced in Yangchun into sell as that were. Drug enterprises usually screen big and plump ones for exporting.
1.2Research of quality index for product specification grade of Fructus Amomi
Learning from the idea and methods for sensorial evaluating agricultural and other commodities, Delphi method was applied to sensorial evaluate3attributes index "weight, diameter, longitudinal diameter" for screening commodity Fructus Amomi. Sensorial characteristics items of the commodity were designed to determine and collect related index for sensorial judgment. The "Data management system for commodity Fructus Amomi" was developed, combining with computer database technology. Experiments were carried out to study sensorial attributes index and their relationship. Evaluating result of the experts and data of the corresponding samples'3attributes index "weight, diameter, longitudinal diameter" were analyzed to find out that grade score experts gave was highly related to the fruit diameter and weight. Then we used robust regression method to generate a prediction model and parameter fitting linear regression equation, to gain a formula calculating Fructus Amomi's grade and trait differences between grades. In a word, this study established product specification grade for commodity Fructus Amomi at3levels.
Exclusive traits of commodity Fructus Amomi:dried; oval or orbicular-ovate, unobvious triangular; brown cutis, density spike-like protuberance, residual perianth on the top; pericarp thin and soft; seed assembled into groups, with three blunt edge, white septum in the middle divide the seed group into3valves; seeds irregular polyhedron, reddish brown or dark brown on the surface, with fine wrinkles, light brown membranous aril on the outside, hard, endosperm gray; strong fragrant; spicy, cool, slightly bitter; no impurities, insects and mildew.
1st grade:fruit weight>lg, fruit longitudinal diameter>17mm, fruit diameter>14mm; each valve of seed group16-24, seed diameter2-3mm.
2nd grade:fruit weight0.55-1g, fruit longitudinal diameter14-17mm, fruit diameter12-14mm; each valve of seed group12-15, seed diameter2-3mm.
3rd grade:fruit weight<0.55g, fruit longitudinal diameter<14mm, fruit diameter<12mm; each valve of seed group7-11, seed diameter2-3mm.
1.3Comparative study of different product specification of Fructus Amomi by chemical analyze and bioassay.
Determination of the efficacy ingredients, metabolites, pesticide residues and heavy metals of Fructus Amomi were carried out according to the "Chinese Pharmacopeia". Relationship between chemical composition and product specification grade was analyzed, results showed as bellowed:
1.3.1Volatile oil content of different grades of Fructus Amomi met the requirements of "Chinese Pharmacopeia"(2010vision). Differences of volatile oil content between levels were not obvious.
1.3.2Bornyl acetate content of different grades of Fructus Amomi met the requirements of "Chinese Pharmacopeia"(2010vision), among which1st grade>2nd grade>3rd grade. Method of "Eucidean Distance Squared" in the "Hierarchical Cluster Analysis" was applied to do cluster analysis on the bornyl acetate content of different grades of Fructus Amomi. It turn out to be that the1st grade tend to be a separate class, with the2nd and3rd grade to be a polymerized class. It mean that bornyl acetate content of the1st grade's was highest, while that of the2nd and3rd grade were of no differences.
1.3.3Contents of water, poison heavy metals and pesticides residue of different grades of Fructus Amomi met the requirements of "Chinese Pharmacopeia"(2010vision).
1.3.4GC-MS analysis results revealed that compositions of volatile oil in Fructus Amomi ordered by their relative contents were bornyl acetate, camphor, d-limonene, borneol, camphene, and others. Among the17compounds, bornyl acetate added up to45%of the total essential oil. No significant differences showed among the three grades of Fructus Amomi in their volatile oil compositions, according to systematic cluster analysis. However, the average percentage contents showed that1st grade>2nd grade>3rd grade.
1.3.5Results of glacial acetic acid induced writhing experiment indicated that the volatile oil of three grades of Fructus Amomi all showed analgesic effect. Among them, the1st grade showed rapid onset of analgesia and longer maintain time, with an analgesia effect from10min to60min, gavage at20ml/kg at14%concentration. The2nd and3rd grade's analgesia effect were unstable, relatively. Gavage at14%concentration, the2nd grade showed analgesia effect80min after administration, while the3rd grade become effective at80-160min, with no differences between them.
1.4Founding of product specification grade standard for the drug Fructus Amomi (draft)
Product specification grade standard for the drug Fructus Amomi (draft) was set up according to the current "Chinese Pharmacopeia" and our research results. It's a draft industrial standard for Fructus Amomi, with the main contents of "original identification-character identification-physical and chemical identification-bioassay-normal detection", including original, processing method, character, product specification grade, identification, detection, determination, biological potency test and storage.
2Conclusion
Learning from evaluating method for product specification of agricultural product, we analyzed the experiment results and created models to predict classification for Fructus Amomi, combining with computer database technology. Furthermore, we investigated if it's scientific and rational to classify product specification grade for Fructus Amomi, by "product specifications-chemical composition-biological effect" correlation analyzing. In this study, we set standard for product specification grade of Fructus Amomi, based on systematic quality index. This study provided reference for product specification grade of fruit groups of TCM.
This study enriched technical content of "geographical indication" of Fructus Amomi, provided scientific guidance for the harvest, processing, storage and marketing in the GAP process of Fructus Amomi in the generating district. It also provided basic for managing policy and help setting the industrial standard for Fructus Amomi.
砂仁来源于姜科植物阳春砂Amomum villosum Lour.、绿壳砂Amomum villosum Lour. var. xanthioides T. L. Wu et Senjen,或海南砂Amomum longiligulare T. L. Wu的干燥成熟果实,是我国著名“四大南药”之一。目前我国市场流通的砂仁主要以阳春砂为主,都是以“统货”进行收购、加工和销售,无规范的商品规格及其行业标准。本研究采用文献整理、现场调研等溯源式研究,分析阳春砂药材商品规格的形成模式,通过理化实验结合道地性形成模式研究,构建“基原鉴定—性状鉴定—理化鉴定—生物检定—常规检测”为主要内容的阳春砂商品规格行业标准草案,为阳春砂商品规格等级划分的客观化、规范化、标准化提供科学依据,为提高和规范中药材商品规格等级评价标准提供技术方法参考。
一、研究内容和主要研究结果
(一)阳春砂文献调研、道地药材产区的现场调研与取样
1、通过文献调研及对阳春砂主产区的现场调研,以及中药材专业市场、药材公司等的调研和咨询,发现目前市场上“阳春砂”均以“统货”进行收购、销售,不分等级,尚未有不同等级阳春砂的科学分类标准和行业质量标准。
2、文献及主产区调研结果表明,道地药材阳春砂生长需要特定的自然生态环境条件,特别与种植地的湿度、温度、日照、土壤等生态因子密切有关。由于栽培地生态条件的不断变化、栽培技术上的差异以及种质自身的变异,在产区也形成了一些栽培品种(如广东阳春市的“长果”、“圆果”和选育中品种“春选”,云南西双版纳的“高杆”、“中杆”、“低杆”等)。
3、虽然广东省阳春市制订了阳春砂的分级(农业)标准,但当地在收购、销售阳春砂时,并未按照分级标准进行,均以“统货”进行收购、销售。此外,由于市场长期认为广东省阳春市产的阳春砂质量好,因此阳春产的阳春砂药材价格高(300-500元/千克),收购外地产阳春砂加工成干果作为阳春产阳春砂销售的情况存在,也导致在药材市场上有些商家将“非阳春产阳春砂”当作是“阳春产阳春砂”药材进行销售。药材公司通常会挑选个大、饱满度较好的阳春砂供应出口。
(二)阳春砂不同商品规格等级定量化指标的研究
借鉴农产品等商品的感观评价思路和方法,采用特尔菲(Delphi)法筛选出阳春砂商品感官评价的“重量、横径、纵径”三个属性指标；设计阳春砂药材商品感官特征项目,测量和收集与阳春砂商品感官判断相关的指标,结合计算机数据库技术,研究开发了“阳春砂药材商品数据管理系统”。进行感官属性相关度研究,对专家等级评判结果与相应阳春砂样品的果实横径、纵径、重量三个属性间的数值关系进行数据分析实验,得出专家等级评分与果实横径、重量相关性最高。使用稳健回归方法产生预测模型,加上参数拟合线性拟合方程得出了阳春砂不同等级的计算公式,得出了一、二、三等品阳春砂的商品规格等级性状差异,得出样品等级计算的线性方程。研究建立了一、二、三等品阳春砂的商品规格等级。
阳春砂商品专属性状：干货。呈椭圆形或卵圆形,有不明显的三棱。表面棕褐色,密生刺状突起,顶端有花被残基。果皮薄而软。种子集结成团,具三钝棱,中有白色隔膜,将种子团分成3瓣。种子为不规则多面体,表面棕红色或暗褐色,有细皱纹,外被淡棕色膜质假种皮；质硬,胚乳灰白色。气芳香而浓烈,味辛凉、微苦。无杂质、虫蛀、霉变。
一等品阳春砂：果实重量1克以上,果实纵径17毫米以上,果实横径14毫米以上。种子团每瓣有种子16-24粒。种子直径2-3毫米。
二等品阳春砂：果实重量在0.55-1克之间,果实纵径在14-17毫米,果实横径在12-14毫米之间。种子团每瓣有种子12-15粒。种子直径2-3毫米。
三等品阳春砂：果实重量0.55克以下,果实纵径14毫米以下,果实横径12毫米以下。种子团每瓣有种子7-11粒。种子直径2-3毫米。
(三)基于化学分析和生物检测的阳春砂不同商品规格比较研究
基于《中国药典》标准的药效指标成分含量测定、阳春砂药材代谢产物、农残、重金属等检测、化学成分含量高低与商品规格等级的相关性、化学成分含量比例与商品规格等级的相关性,以及基于生物检测的阳春砂药材商品规格比较研究结果如下。
1、各等级商品阳春砂挥发油含量均符合《中国药典》(2010年版一部)规定,不同等级间的阳春砂挥发油含量差异不大。
2、各等级商品阳春砂乙酸龙脑酯含量均符合《中国药典》(2010年版一部)规定,乙酸龙脑酯的含量是一等品>二等品>三等品。采用“层次聚类分析方法”中的欧式距离平方对三个等级阳春砂的乙酸龙脑酯进行聚类分析,二等品和三等品聚为一类,一等品单独为一类,即二等品和三等品的乙酸龙脑酯含量无明显差异,一等品显著高于其他两个品级。
3、各等级商品阳春砂水分含量、有害重金属、农药残留量均符合《中国药典》(2010年版一部)规定。
4、阳春砂挥发油的气质联用色谱分析结果表明,阳春砂挥发油主要峰相对百分含量排序依次为乙酸龙脑酯、樟脑、d-柠檬烯、冰片、莰烯等共十七种挥发性化合物,其中乙酸龙脑酯为阳春砂挥发油中的主要成分,占挥发油总量的45%以上。从系统聚类结果分析,各等级阳春砂挥发油化学组成与相对含量无明显的差异,但挥发油中各成分平均百分比含量结果为一等品>二等品>三等品。
5、以冰乙酸致小鼠扭体实验为生物效价指标的研究结果表明,各等级阳春砂挥发油均有镇痛作用,其中一等品阳春砂镇痛的量效、时效关系比较明确,14%浓度一等品阳春砂挥发油以20mL/kg灌胃,从10min至160min均有镇痛作用,提示一等品镇痛起效较快,维持时间较长。二等品、三等品阳春砂挥发油的镇痛药效相对不稳定,14%浓度二等品阳春砂挥发油在给药80min时对冰乙酸致小鼠扭体有镇痛作用,14%浓度三等品阳春砂挥发油在给药80min和160min时有镇痛作用,两者无差异。
(四)阳春砂药材商品规格等级标准(草案)的建立
参照现版《中国药典》和研究结果,起草阳春砂药材商品规格等级标准(草案),主要包括基原、加工方法、性状、商品规格等级、鉴别、检查、含量测定、生物效价检测、贮藏等方面,形成“基原鉴定—性状鉴定—理化鉴定—生物检定—常规检测”为主要内容的阳春砂商品规格行业标准草案。
二、研究结论
本论文通过参考和借鉴农产品等商品规格评价的方法,结合计算机数据库技术,对实验数据进行分析建模实现对阳春砂商品规格的分级预测,并进一步通过“商品规格—化学成分—生物效应”的关联分析,探讨了阳春砂药材商品规格等级的科学合理性。研究建立了基于系统量化指标的阳春砂药材商品规格等级标准,研究方法可为果实类中药材的商品规格等级研究提供参考和借鉴。
本研究丰富了阳春砂“地理标志产品”商品规格的技术内涵,可为阳春砂道地产区阳春砂GAP过程中的采收、加工、贮藏和商品销售提供科学指导,为阳春砂优质优价提供依据,为进一步制订阳春砂行业标准和管理政策提供依据。
Product specification is a qualitative reflection of Traditional Chinese Medicine (TCM). It's the most intuitive indicator to evaluate TCM."Product Specification Standard of76Drugs", the first norm for product specification of TCM promulgated and implemented in China1984, set classification and standard on product type, specification and grade of76common TCM, according to those drugs' quality. However, with TCM's continuous shifting of product manufacturing and circulation, as well as their renewing of quality standard, this norm does not correspond with the current manufacturing, circulation and apply of TCM. In the current TCM market, no unitive quality standard is acknowledged by the druggists. Because of the complicated product specification and grade of some drugs, the failure of inheriting traditional discerning experiences, and the absence of scientific and systematic technical information, product specification standard cannot play its role in the quality standard evaluating system of TCM's product specification. Therefore, it's necessary to regulate and improve the technology in TCM's product specification evaluating system, and to set a qualitative standard for TCM's product specification and grade. In this study, an integrated evaluating method is established to form and evaluate the industrial standard for the drug Fructus Amomi, based on systematic quality index.
One of China's well-known "Four Southern Medicine", Fructus Amomi, comes from dry mature fruit of Zingiberaceae plants, including Amomum villosum Lour., Amomum villosum Lour. var. xanthioides T. L. Wu et Senjen and Amomum longiligulare T. L. Wu. The main circulation specie in the current TCM market is Amomum villosum Lour.. It's purchased, processed and distributed as unitive goods, without regulating product specification and industrial standard. In this study, traceable research methods, such as literature reviewing and site investigating, were applied to analyze the formation model of Fructus Amomi's product specification. By physical and chemical experiments, together with authentic formation model study, a draft industrial standard for Fructus Amomi's product specification was established to provide scientific basis for objective, norm and standardized classification of Fructus Amomi's product specification and grade. This standard was based on the "original identification-character identification-physical and chemical identification-bioassay-normal detection". Therefore, this study provides technic reference to regulate and improve evaluating standard for TCM's product specification and grade.
1Research contents and main findings
1.1Literature review, site investigation and sample collection of Fructus Amomi
1.1.1Through literature reviewing, site investigating and investigating on TCM market and enterprises, we found that Fructus Amomi was purchased, processed and distributed as unitive goods, without a scientific classifying standard and industrial standard for different grade of the product.
1.1.2Results of literature reviewing and site investigating showed that it needs special nature environmental conditions, especially ecological factors, such as humidity, temperature, sunlight and earth, for Fructus Amomi to grow well. Due to the continuous variation of ecological conditions and cultivable technology, as well as the mutation of the specie itself, new cultivable species could be formed in the generating districts.("Long fruit","round fruit","spring selected" species in Yangchun City, Guangdong province, and "high trunk","medium trunk" and "short trunk" in Xishuangbanna, Yunnan province.)
1.1.3Although classification standard for Fructus Amomi had been made by Yangchun City, Guangdong province, Fructus Amomi were purchased and processed as unitive goods, without classifying according to the standard. Besides, because Fructus Amomi produced by Yangchun City was consumed the best in the TCM market, its price(300-500￥/kg) is higher compared to other species. Illegal phenomenon do exist that some druggists put Fructus Amomi not produced in Yangchun into sell as that were. Drug enterprises usually screen big and plump ones for exporting.
1.2Research of quality index for product specification grade of Fructus Amomi
Learning from the idea and methods for sensorial evaluating agricultural and other commodities, Delphi method was applied to sensorial evaluate3attributes index "weight, diameter, longitudinal diameter" for screening commodity Fructus Amomi. Sensorial characteristics items of the commodity were designed to determine and collect related index for sensorial judgment. The "Data management system for commodity Fructus Amomi" was developed, combining with computer database technology. Experiments were carried out to study sensorial attributes index and their relationship. Evaluating result of the experts and data of the corresponding samples'3attributes index "weight, diameter, longitudinal diameter" were analyzed to find out that grade score experts gave was highly related to the fruit diameter and weight. Then we used robust regression method to generate a prediction model and parameter fitting linear regression equation, to gain a formula calculating Fructus Amomi's grade and trait differences between grades. In a word, this study established product specification grade for commodity Fructus Amomi at3levels.
Exclusive traits of commodity Fructus Amomi:dried; oval or orbicular-ovate, unobvious triangular; brown cutis, density spike-like protuberance, residual perianth on the top; pericarp thin and soft; seed assembled into groups, with three blunt edge, white septum in the middle divide the seed group into3valves; seeds irregular polyhedron, reddish brown or dark brown on the surface, with fine wrinkles, light brown membranous aril on the outside, hard, endosperm gray; strong fragrant; spicy, cool, slightly bitter; no impurities, insects and mildew.
1st grade:fruit weight>lg, fruit longitudinal diameter>17mm, fruit diameter>14mm; each valve of seed group16-24, seed diameter2-3mm.
2nd grade:fruit weight0.55-1g, fruit longitudinal diameter14-17mm, fruit diameter12-14mm; each valve of seed group12-15, seed diameter2-3mm.
3rd grade:fruit weight<0.55g, fruit longitudinal diameter<14mm, fruit diameter<12mm; each valve of seed group7-11, seed diameter2-3mm.
1.3Comparative study of different product specification of Fructus Amomi by chemical analyze and bioassay.
Determination of the efficacy ingredients, metabolites, pesticide residues and heavy metals of Fructus Amomi were carried out according to the "Chinese Pharmacopeia". Relationship between chemical composition and product specification grade was analyzed, results showed as bellowed:
1.3.1Volatile oil content of different grades of Fructus Amomi met the requirements of "Chinese Pharmacopeia"(2010vision). Differences of volatile oil content between levels were not obvious.
1.3.2Bornyl acetate content of different grades of Fructus Amomi met the requirements of "Chinese Pharmacopeia"(2010vision), among which1st grade>2nd grade>3rd grade. Method of "Eucidean Distance Squared" in the "Hierarchical Cluster Analysis" was applied to do cluster analysis on the bornyl acetate content of different grades of Fructus Amomi. It turn out to be that the1st grade tend to be a separate class, with the2nd and3rd grade to be a polymerized class. It mean that bornyl acetate content of the1st grade's was highest, while that of the2nd and3rd grade were of no differences.
1.3.3Contents of water, poison heavy metals and pesticides residue of different grades of Fructus Amomi met the requirements of "Chinese Pharmacopeia"(2010vision).
1.3.4GC-MS analysis results revealed that compositions of volatile oil in Fructus Amomi ordered by their relative contents were bornyl acetate, camphor, d-limonene, borneol, camphene, and others. Among the17compounds, bornyl acetate added up to45%of the total essential oil. No significant differences showed among the three grades of Fructus Amomi in their volatile oil compositions, according to systematic cluster analysis. However, the average percentage contents showed that1st grade>2nd grade>3rd grade.
1.3.5Results of glacial acetic acid induced writhing experiment indicated that the volatile oil of three grades of Fructus Amomi all showed analgesic effect. Among them, the1st grade showed rapid onset of analgesia and longer maintain time, with an analgesia effect from10min to60min, gavage at20ml/kg at14%concentration. The2nd and3rd grade's analgesia effect were unstable, relatively. Gavage at14%concentration, the2nd grade showed analgesia effect80min after administration, while the3rd grade become effective at80-160min, with no differences between them.
1.4Founding of product specification grade standard for the drug Fructus Amomi (draft)
Product specification grade standard for the drug Fructus Amomi (draft) was set up according to the current "Chinese Pharmacopeia" and our research results. It's a draft industrial standard for Fructus Amomi, with the main contents of "original identification-character identification-physical and chemical identification-bioassay-normal detection", including original, processing method, character, product specification grade, identification, detection, determination, biological potency test and storage.
2Conclusion
Learning from evaluating method for product specification of agricultural product, we analyzed the experiment results and created models to predict classification for Fructus Amomi, combining with computer database technology. Furthermore, we investigated if it's scientific and rational to classify product specification grade for Fructus Amomi, by "product specifications-chemical composition-biological effect" correlation analyzing. In this study, we set standard for product specification grade of Fructus Amomi, based on systematic quality index. This study provided reference for product specification grade of fruit groups of TCM.
This study enriched technical content of "geographical indication" of Fructus Amomi, provided scientific guidance for the harvest, processing, storage and marketing in the GAP process of Fructus Amomi in the generating district. It also provided basic for managing policy and help setting the industrial standard for Fructus Amomi.
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