Volpi Nicola, Bergonzini Gianluca
Department of Biologia Animale, University of Modena and Reggio Emilia, Via Campi 213/d, 41100 Modena, Italy.
J Pharm Biomed Anal. 2006 Sep 26;42(3):354-61. doi: 10.1016/j.jpba.2006.04.017. Epub 2006 Jun 9.
In this paper, the qualitative and quantitative separation and determination of the polyphenolic component of propolis preparations in the form of ethanolic extract, usually used for commercial pharmaceutical preparations, has been investigated by means of on-line HPLC-ESI/MS technique. Propolis of different origin have been evaluated for their components and a specific fingerprint has been determined potentially useful for the quality control of extracts in pharmaceutical preparations. The ethanolic extracts of propolis from Argentina, Italy and Spain shows approximately the same total ion chromatogram (TIC) profile due to the presence of the same molecular species, identified by the negative ESI-MS. On the contrary, the samples from Azerbaijan, China, Ethiopia and Kenya show a very peculiar TIC profiles. By using many purified flavonoids and calibration curves over a wide concentration range, from 0.05 (5 microg/ml) to 5 microg (500 microg/ml), an accurate assessment of the contents of several bioactive compounds in extract samples was performed. The propolis from Argentina, Italy and Spain show a great amount of pinocembrin (approximately 49%, 48% and 39% of the total identified flavonoids, respectively) and variable but similar percentages of the other species. On the contrary, the propolis from China, Azerbaijan and Ethiopia have a great amount of pinocembrin (approximately 63%, 46% and 62%, respectively) but no presence of genistein, kaempferol, apigenin and chrysin for the sample from China, genistein, kaempferol, acacetin and chrysin for the propolis from Azerbaijan, and no kaempferol and acacetin for the sample from Ethiopia. The ethanolic extract from propolis of Kenya has no identified flavonoid species but just a peak possessing a m/z of 253.0. Finally, an evaluation of the presence of total flavonoids for the various propolis samples was performed, with extracts from Argentina, Italy and Spain more rich in polyphenols than those from Azerbaijan, China, Ethiopia and Kenya. The HPLC-ESI/MS under the experimental conditions illustrated represents a valuable method for the qualitative and quantitative assay of the most relevant components of propolis. On-line HPLC-ESI/MS analysis constitutes an alternative to obtain typical fingerprints of propolis and a reliable identification of a large number of propolis polyphenolic components.
本文采用在线高效液相色谱-电喷雾电离质谱联用（HPLC-ESI/MS）技术，对通常用于商业药物制剂的蜂胶乙醇提取物形式制剂中的多酚成分进行了定性和定量分离与测定。对不同产地的蜂胶成分进行了评估，并确定了一个特定的指纹图谱，该图谱可能对药物制剂中提取物的质量控制有用。由于存在相同的分子种类，通过负离子电喷雾电离质谱（ESI-MS）鉴定，来自阿根廷、意大利和西班牙的蜂胶乙醇提取物显示出大致相同的总离子色谱图（TIC）谱。相反，来自阿塞拜疆、中国、埃塞俄比亚和肯尼亚的样品显示出非常独特的TIC谱。通过使用多种纯化的黄酮类化合物以及在0.05（5微克/毫升）至5微克（500微克/毫升）的宽浓度范围内的校准曲线，对提取物样品中几种生物活性化合物的含量进行了准确评估。来自阿根廷、意大利和西班牙的蜂胶含有大量的松属素（分别约占已鉴定黄酮类化合物总量的49%、48%和39%），其他种类的含量各不相同但相似。相反，来自中国、阿塞拜疆和埃塞俄比亚的蜂胶含有大量的松属素（分别约为63%、46%和62%），但中国的样品中没有染料木黄酮、山奈酚、芹菜素和白杨素，阿塞拜疆的蜂胶中没有染料木黄酮、山奈酚、刺槐素和白杨素，埃塞俄比亚的样品中没有山奈酚和刺槐素。来自肯尼亚的蜂胶乙醇提取物中没有鉴定出黄酮类化合物种类，只有一个质荷比为253.0的峰。最后，对各种蜂胶样品中总黄酮的存在情况进行了评估，来自阿根廷、意大利和西班牙的提取物比来自阿塞拜疆、中国、埃塞俄比亚和肯尼亚的提取物含有更多的多酚。在所示实验条件下的HPLC-ESI/MS是一种用于蜂胶最相关成分定性和定量分析的有价值方法。在线HPLC-ESI/MS分析是获得蜂胶典型指纹图谱以及可靠鉴定大量蜂胶多酚成分的一种替代方法。
