基于抗氧化“活性指纹图谱”的蜂胶质量控制研究

Propolis is the hot topic in bee products research and development in recent 10 years. The chemical composition of propolis various greatly from different areas, the qualities and effect of prepared propolis sold in market must be payed attention to their divergences. To clarify the pharmacodynamic substance foundation and for controling accurately and efficiently of the propolis quality, correlation studies focused on the chemical composition and pharmacological activities of propolis are necessary. The spectrum-effect relationship research of propolis was performed based on antioxidant activities and chromatographic fingerprint of flavonoids and phenolic acids. The antioxidant-related active ingredients were screened by developing the HPLC-DPPH method and analyzing the relationship between chromatographic fingerprint and efficacy based on chemometricsto. The “antioxidant effective fingerprint” was developed containing enough antioxidant active components, which were separated and identified by HPLC, MS, NMR etc. Combination of the in vitro antioxidant activities, the content of total phenol and total flavonoids to evaluate the effective of developed antioxidant "effective fingerprint", we will develop the quality evaluation method based on antioxidant "effective fingerprint". This project highlights the unique material foundation of propolis, provides a new idea for in-depth development of fingerprint efficacy of propolis and important theoretical basis to guarantee the stable pharmacodynamics and improve the propolis quality control standards.

蜂胶是近十年来蜂产品研究与开发的热点。由于不同来源的蜂胶化学组成差异较大，致使产品质量良莠不齐、药效难以稳定发挥，亟需开展蜂胶化学成分组成与药理活性相关性研究，阐明蜂胶的药效物质基础，准确、有效地控制蜂胶的质量。本项目拟以抗氧化活性为切入点开展蜂胶谱-效物质基础研究，针对黄酮类和酚酸类有效部位，建立化学指纹图谱；建立HPLC-DPPH快速筛选抗氧化活性特征峰的方法，结合化学计量学研究蜂胶谱效关系，选定抗氧化活性指标成分，并采用HPLC、MS、NMR等多项技术进行解析；对活性指标成分进行效价计算，建立抗氧化“活性指纹图谱”；结合抗氧化活性、总酚及总黄酮含量综合评价构建的“活性指纹图谱”的有效性，建立蜂胶抗氧化“活性指纹图谱”质量评价方法。本项目的研究凸显蜂胶独具特色的抗氧化物质基础，为深入开展蜂胶指纹图谱的谱效学研究提供了新思路，为保证蜂胶药效稳定，完善蜂胶质量控制标准奠定了的理论基础。

蜂胶是近十年来蜂产品研究与开发的热点。由于不同来源的蜂胶化学组成差异较大，致使产品质量良莠不齐、药效难以稳定发挥，亟需开展蜂胶化学成分组成与药理活性相关性研究，阐明蜂胶的药效物质基础，准确、有效地控制蜂胶的质量。本项目以抗氧化活性为切入点开启蜂胶谱-效物质基础研究，针对黄酮类和酚酸类有效部位，分别建立了中国蜂胶和巴西绿蜂胶化学指纹图谱；建立了HPLC-DPPH快速筛选抗氧化活性特征峰的评价方法，结合化学计量学研究蜂胶的谱效关系，选定抗氧化活性指标成分，并采用HPLC-MS技术进行解析，从中国蜂胶中选定12个具有抗氧化活性的特征峰：咖啡酸、阿魏酸、槲皮素、山奈酚、芹菜素、咖啡酸异戊二烯酯、咖啡酸苄酯、短叶松素-3乙酸酯、CAPE、高良姜素、6-甲氧基柯因、p-香豆酸苄酯；从巴西绿蜂胶中选定9个抗氧化活性指标成分：咖啡酸、绿原酸、3,5-二咖啡酰奎宁酸、4,5-二咖啡酰奎宁酸、3,4,5-三咖啡酰奎宁酸、山奈酚、山奈素、桦木酚及阿替匹林C；建立了《蜂胶中杨树胶的检测方法-高效液相色谱法》（GB/T 34782-2017）、《蜂胶中阿替匹林C的测定方法-高效液相色谱法》（GH/T 1114-2015）、《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3个标准以蜂胶中酚酸和黄酮类的定性和定量检测来评价蜂胶的真实性和质量优劣，标准的颁布与实施，必将为净化蜂胶市场，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强有力的技术支撑。本项目的研究凸显蜂胶独具特色的抗氧化物质基础，为深入开展蜂胶指纹图谱的谱效学研究提供了新思路，为保证蜂胶药效稳定奠定了的理论基础，完善了蜂胶质量控制标准，为蜂胶行业的健康发展提供了检测依据。