【作者】 郭丽丽；

【导师】 葛毅强； 陈颖；

【作者基本信息】 中国农业大学 ， 农产品加工及贮藏工程， 2014， 博士

【摘要】 燕窝是由金丝燕及多种同属燕类所筑的巢窝,主产于马来西亚、印度尼西亚、菲律宾、越南等东南亚国家和地区,自古以来一直被视为一种名贵中药和珍稀食品。近年来,燕窝在我国的消费量呈逐年上升趋势,与此同时,燕窝的进口价格也在逐渐攀升。受巨额利润的驱使,市场中燕窝的质量安全状况不容乐观,燕窝的掺假掺杂现象严重。另一方面,不同产地、不同生产方式的燕窝产品价位悬殊,利润空间巨大,燕窝的以次充好现象严重。由于缺乏相关的产品认证制度和溯源监管体系,致使目前市场上的燕窝产地、生产方式标识不清,严重损害了消费者和燕窝企业的权益。此外,燕窝产地及生产方式的判别也是行业分类标识的需求。因此,亟待加强对燕窝产品原料及来源的真伪鉴别,建立可用于燕窝中掺假物以及燕窝产地和生产方式的快速、准确鉴别方法。本论文研究了燕窝及其掺假物成分和不同产地及生产方式燕窝的鉴别方法,具体研究工作如下：1.针对燕窝质量快速初筛的需要,对不同来源的燕窝样品、燕窝及其掺假物和燕窝与掺假物的混合样品进行了红外光谱分析研究,将所得的光谱数据进行主成分分析,初步建立了燕窝真伪的判别模型。该模型可鉴别出较高含量掺假燕窝(掺假量≥30%)的真伪,但无法准确识别出掺假燕窝中的银耳或琼脂成分,适合于对燕窝产品真伪的快速鉴别和对燕窝产品质量的初步筛查。2.为了对燕窝中的掺假物成分进行准确定性,选取核酸作为研究对象,通过筛选和设计燕窝及其常见掺假物成分的特异性引物和探针,建立了基于分子生物学的燕窝及其掺假物的实时荧光PCR及液相芯片检测方法。对市售燕窝样品的检测结果表明所建立的实时荧光PCR和液相芯片体系灵敏度高,可用于对燕窝样品中掺假物成分的准确定性检测。3.针对燕窝产地及生产方式的鉴别,选取蛋白质组作为研究对象,采用SDS-PAGE电泳技术分离了不同燕窝样品的蛋白质,并将所得的蛋白条带数据进行多变量分析,初步建立了燕窝产地及生产方式的判别模型。采用双向电泳技术比较了不同燕窝样品的蛋白质组,筛选出不同生产方式与产地的差异蛋白点,为后续质谱鉴定奠定了基础。综上所述,本论文利用红外检测、基因检测及蛋白质组学分析技术,可分别实现对燕窝产品真伪的快速筛查、燕窝中掺假物成分的准确定性及燕窝产地及生产方式的判别,从而初步建立了燕窝质量检测的表征属性识别技术体系,该体系也可满足条件不同机构和不同检测目的的需求,为监控燕窝产品、整顿燕窝市场和规范燕窝行业提供技术支撑和理论指导,同时也为我国正致力于建立的燕窝监管溯源管理体系提供思路。更多还原

【Abstract】 Edible bird’s nest (EBN) is a kind of nests constructed by the swiftlet of Aerodramus genus and other closely related species, which mainly inhabited in Southest Asia countries such as Malaysia, Indonesia, Philippine, Vietnam. EBN has been regarded as a precious and valuable medical food for a long time. In recent years, the EBN comsumption has been growing. Driven by huge profits, a large number of EBNs with unidentified origins are input in an improper way by immoral retailers. The status of EBN quality and safety in market is unoptimistic and the phenomenon of adulteration in EBN with cheap materials is severe. On the other hand, the prices of EBNs with different geographic origin or production mode are various with huge profits, resulting in the severe phenomenon of shoddy. Nevertheless, the lack of related product certification system and traceability supervision system incur such phenomenons of unfair competition as ambiguous origin labeling and origin mislabeling, which seriously destroy the benefits of consumers and EBN enterprises. Therefore, it is urgent to strengthen the quality detection of EBN products and develop the rapid and accurate method for identifying adulterants from EBNs and the origin of EBNs.This paper studied on the method for authenticating EBNs and discriminating the EBN origins (geographic origin and production mode). The specific studies are as follows:1. In order to meet the requirement of rapid quality detection for EBN products, infrared spectroscopy was utilized to detect EBNs with different origins, the separate samples of EBN and adulterants and the mixed samples of both. The obtained infrared spectra data were conducted by Principle Component Analysis and a discrimination model for authentication of EBNs was preliminarily established. The model could be used for screening the adulterated EBNs with the quantity of adulterants above30%(w/w). However, it failed to distinguish the specific adulterant component from the adulterated EBNs with white fungus or agar. The results showed that infrared spectroscopy could be effectively used for rapid screening of EBN authentication and preliminary monitoring of EBN quality.2. In order to accurately detect the adulterant ingredients in EBNs, nucleic acid was selected as the object, and specific primers and probe for EBN and adulterant components were screened and designed to establish the method of the real-time PCR and suspension bead array based on the molecular biology. The detection results of commercial EBN samples suggested that the established real-time PCR and suspension bead array with high sensitivity can be used for accurately identifying the adulterant ingredients in EBN products.3. In order to study on the discrimination method of EBN origins, the protein was selected as the object and SDS-PAGE was used for analyzing the proteins in different EBN samples. Then multivariate analysis was conducted based on the obtained data from protein bands and points, and finally the discriminant model was preliminarily established for the determination of geographic origin and production mode for EBNs. Then two-dimensional electrophoresis (2-DE) was utilized to compare the proteomes of different EBNs and the protein points representing different production modes or geographic origins were screened out, which lays a foundation for subsequent protein identification by mass spectrum.In summary, this study combined such three technologies as the infrared detection, genetic testing and protein analysis technique for preliminarily establishing a comprehensive set of quality detection system for EBN products based on the representative characteristics recognition technology, which involved in the rapid screening for EBN quality, qualitative detection of adulterants components and discrimination of geographic origin and peoduction mode for EBNs. The developed detection system can meet the requirements of various institutions for different purposes. The study in this thesis could provide technical support and therotical guidance for positively monitoring EBN products, supervising EBN market and regulating EBN industry, and also could provide ideas for the establishement of EBN supervision traceability management system conducted by our country.更多还原