基于多指标成分定量结合指纹图谱定性的蜂胶真实性评价体系的构建及机理研究

Propolis is a natural remedy that has been employed extensively since ancient times. In recent decades, propolis has been used in popular medicine, health food,and everyday products. The yield of propolis may vary considerable, depending on several factors including honeybee species, climate, and plant sources.The high demand and limited availability for propolis lead to gross adulteratation. Poplar tree's gum, the extract of populus buds, or artificially adding some flavonoids to enhance the content of total flavonoids of propolis are the major false way, has become the major bottlenecks to restrict the healthy development of propolis industry. The project is planned to start from the comparative study of the chemical composition of propolis and plant source, screening marker substances of poplar type propolis by HPLC, MS, IR, UV, NMRI etc., achieving the chemical standardization of poplar type propolis based on fingerprint analysis and multi-marker ingredients determination, developing the representative standard HPLC fingerprint of poplar type propolis and poplar tree gum. Finding the characteristic components of poplar tree gum which are absent in propolis and the mechanism of difference, developing the evaluation system for distinguishing propolis from poplar tree gum. Studing the difference of biological activity and physicochemical properties betewwn propolis and poplar gum. Developing evaluation system of propolis authenticity to avoid adulterated occurrence. This project will provide theoretical support and technical guidance to improve the quality standards and effective monitor the propolis quality.

蜂胶自古就被用于天然药物，现已广泛应用于医药、保健食品、日化等行业。但蜂胶生产受蜂种、气候、植物源等因素限制，产量相对有限。受利益驱使，近年来市场上出现了大量的假冒蜂胶。以杨树芽熬制的杨树胶或加入类黄酮人为提高总黄酮含量是蜂胶主要的使假方式，已严重阻碍蜂胶行业的健康发展，亟需建立蜂胶真实性评价体系。本项目拟采用HPLC、MS、IR、UV、NMRI等技术手段，从蜂胶与植物源化学成分异同点入手，筛选出杨树型蜂胶标志性成分，建立多个标志性成分定量结合指纹图谱定性方法实现其化学成分的标准化，建立杨树型蜂胶和杨树胶的HPLC标准图谱；找到杨树胶中特有而蜂胶中不含有的特征成分，探明差异机理，建立鉴别蜂胶与杨树胶的评价体系；从生物学活性、理化特性等角度研究蜂胶与杨树胶的差异；建立蜂胶真实性评价体系，从根源上避免掺杂使假现象的发生。本项目的研究将为完善蜂胶质量标准，有效监控蜂胶质量提供理论支持和技术指导。

蜂胶是一种自古以来就被广泛应用的天然药物，已广泛用于医药保健、食品、日化等行业。但蜂胶生产受蜂种、气候、植物来源等因素限制，产量相对有限。受利益的驱使，市场上出现大量的掺杂使假蜂胶。以杨树芽熬制的杨树胶和（或）加入类黄酮类人为提升总黄酮含量是目前蜂胶主要的使假方式，已成为蜂胶行业健康发展的主要瓶颈。本项目从蜂胶与植物源化学成分对比研究入手，建立了蜂胶中12种黄酮和8种酚酸类化合物的HPLC分析方法，筛选出了杨树型蜂胶的标志性成分，建立了杨树型蜂胶和杨树胶的标准图谱；找到杨树胶中特有而蜂胶中不含有的特征成分-水杨苷和邻苯二酚，并探明了差异机理；从体外自由基清除能力、RAW264.7细胞模型抗氧化能力评价、细菌脂多糖诱导小鼠急性肺损伤模型等角度研究蜂胶与杨树胶的生物学活性差异；建立了中国蜂胶标准化评价方法和蜂胶与杨树胶鉴别方法相结合的蜂胶保真去伪真实性评价体系，从根源上避免掺杂使假现象的发生。本项目建立了《蜂胶中杨树胶的检测方法—反相高效液相色谱法》国家标准和《蜂胶》国家标准、《蜂胶中咖啡酸、p-香豆酸、阿魏酸、短叶松素、松属素、短叶松素3-乙酸酯、白杨素和高良姜素含量的测定—反相高效液相色谱法》行业标准，这三个标准的颁布实施为净化蜂胶市场，维护广大养蜂者、蜂产品生产企业和消费者的合法权益提供了强有力的技术支撑。