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1、河南大学 硕士学位论文 蜂胶和蜂蜜的成分研究 姓名：毛文岭 申请学位级别：硕士 专业：分析化学 指导教师：刘绣华 2011-05 I 摘摘 要要 本文以河南省驻马店地区产的蜂胶(propolis)和五种不同花源蜂蜜(honey)为研究对 象，着重对蜂胶与五种蜂蜜中总黄酮、总酚酸等活性成分和醛类、醇类等挥发性成分进 行分析，并且对蜂胶中铅的含量进行测定。其中总黄酮含量测定采用三氯化铝比色法进 行，总酚酸的含量采用 Folin-Ciocalteu 比色法，挥发性成分采用气相色谱-质谱联用 (GC-MS)的方法，而对蜂胶中铅的测定建立了一种成本低廉、效果满意的紫外分光光度 法。本论文主要内容如下：
2、.蜂胶中挥发性成分、活性成分及重金属含量测定蜂胶中挥发性成分、活性成分及重金属含量测定 对蜂胶中挥发性成分的分析，在分析样品制备时，分别采用了溶剂萃取法和固相萃 取法进行样品预处理。前者分别采用了二氯甲烷、乙腈及乙酸乙酯进行液液萃取；后者 以自制活性炭固相萃取柱对蜂胶挥发性成分进行固相萃取(SPE)，结果显示，用乙酸乙 酯与固相萃取的方法得到的挥发性成分中酯类物质所占比例较大， 有机酸类化合物化合 物较少。用二氯甲烷与乙腈萃取的方法中得到的醇类化合物较为丰富。 采用 AlCl3比色法对蜂胶中总黄酮的含量进行测定，对照品和样品在 440nm 处均有 最大吸收，线性回归方程为 Y=0.01465+
3、0.03603X (r =0.9994)，测得蜂胶中总黄酮含量平 均值为 3.753%。用 Folin-Ciocalteu 比色法测定蜂胶中的总酚酸含量，线性回归方程为： Y=0.00982+0.06965X (r =0.9992)，测得蜂胶中总酚酸含量为 7.524%。 研究了在碱性条件下，以双硫腙-乙醇-Tween80 为显色剂，在水相反应体系中直接 用双峰双波长法测定铅含量的新方法。显色剂用量选 1.50 mL，pH 值在 8.0-9.0 之间时 吸光度值较高且稳定，线性回归方程为：Y=0.1778X+0.002(r=0.9996)，用于蜂胶中铅的 的测定，结果为 3.571mgkg-1
4、。 蜂蜜中挥发性成分、活性成分测定蜂蜜中挥发性成分、活性成分测定 采用固相萃取制备分析样品，对比分析了五种蜂蜜中挥发性成分，结果显示，在荆 花蜜，夏枯草花蜜，枣花蜜，槐花蜜和油菜花蜜五种花蜜中，乙酸异丙酯只有在荆花蜜 中未被检测出；甲基苄基苯酚、异戊醇、乙酸正丁酯、乙酸异丁酯在枣花蜂蜜中未被检 测出；对甲氧基苯乙醇在槐花蜜中未被检测到；9-二十三碳烯、环十五内酯只有在油菜 花蜜中检出，而乙酸苯乙酯只有在油菜蜜中没有；薄荷醇和二十八烷仅在夏枯草蜂蜜中 II 检出；3-甲基环戊二醇、3-甲基-1,3,5-戊三醇只有槐花蜜中没有；检测出乙酸丙酯、丙 酸乙酯、糠醛、安息香醛、二羟基苯乙酮、苯乙醇、邻苯
5、二甲酸二异辛酯等八种物质在 五种蜂蜜中都含有。研究结果为鉴别不同蜜源的蜂蜜提供了参考依据。 采用 AlCl3比色法和 Folin-Ciocalteu 比色法对五种蜂蜜中总黄酮的含量和总酚酸含 量进行了测定，结果为枣花和夏枯草花蜂蜜中的黄酮含量分别为 0.001521%和 0.001375%，槐花蜜，荆花蜜，油菜花蜜用所选择的方法未测出黄酮含量。总酚酸含量 分别为夏枯草蜜 0.06328%，枣花蜜 0.05684%，槐花蜜 0.05565%，荆花蜜 0.05531%， 油菜花蜜 0.05065%。 关键词：蜂蜜，蜂胶，活性成分，铅 III ABSTRACT In this paper, one
6、pecie of propolis and five type of honey which were produced from Zhumadian district of Henan were studied, including total flavonoids, total phenolic acids and volatile components. The concentration of lead in propolis were also determined. Flavonoids were analyzed by aluminum chloride colorimetric
7、 method and the total phenolic content was test through Folin-Ciocalteu colorimetric method. The volatile compounds were determined by gas chromatography-mass spectrometry (GC-MS) method. The determination of lead in propolis by ultraviolet spectrophotometry is proposed with a low- cost, ,environmen
8、t friend and good feasibility. The main contents can introduce detailed as follows: the determination of volatile components,Total flavonoids,Total penolics and the determination of lead in propolis The volatile components in propolis were analysised by GC-MS, on one hand Liquid liquid extraction an
9、d solid phase extraction(SPE) by self-made activated carbon-based SPE column were used to prepare the samples. The results show that the esters obtained in volatile components are larger by ethyl acetate extraction and SPE. the alcohols obtained in volatile components are larger by methylene dichlor
10、ide extraction and acetonitrile extraction. The content of total flavonoids in propolis was determined by AlCl3 photometry at the absorption 440nm. The linear regression equation was Y=0.01465+0.03603 X (r = 0.9994). The average total flavonoid content in propolis is 3.753%. Total phenolic content i
11、n propolis was also detected by Folin-Ciocalteu colorimetry method. The calibration equation is Y = 0.00982 +0.06965 X (r = 0.9992).And the total phenolic content in propolis is 7.524%. A new dual-wavelength peaks method for the determination of lead in water phase reaction system at pH8.0-9.0 by us
12、ing dithizone - ethanol-Tween80 as the color reagent was developed. In this method, selecting 1.50 mL color dosage, pH (8.0-9.0). The calibration equation is Y=0.1778X+0.002 (r=0.9996).The lead content in propolis is 3.571 mgkg-1. The volatile components, Total flavonoids,Total penolics in five type
13、s of Honey The volatile components in the five types of Honey was measured with the same SPE columns as IV in propolis. Comparativly analysis of different volatile components in five types of honey showed that among the Jing nectar honey, nectar Prunella honey, jujube flower Honey, Acacia honey and
14、rape honey, isopropyl acetate only does not exist in the Jing nectar. Benzylphenol, isoamylalcohol, butyl acetate, isobutyl acetate only do not exist in the Jujube flower honey. Metoxybenzene ethanol was not detected in Acacia honey. 9-twentythree carbene, Cyclopentadecanolide were detected only in
15、the rape honey, but phenylethyl acetate does not exist in the rape honey. octacosane and menthol were detected only in the nectar prunella honey honey. 3-methylcyclopentadienyldiol, 3-methyl-1, 3, 5-triol do not exist in Sophorae honey. All the eight components: acetate ethyl, ethyl propionate, furf
16、ural, benzaldehyde, dihydroxyacetophenone, phenylethylalcohol, dibutylphthalate, ethylhexyl phthalate all contain in the five types of honey. Total flavonoids and total phenolic content in honey are also detected by by AlCl3 photometry and Folin-Ciocalteu colorimetry method. The results showed that the flavonoid content in Acacia honey and jujube flower honey is higher than the other three. The average total flavonoid contents in nectar prunella honey is 0.001375%,in jujube flowe
