UPLC-MS/MS法分析洋槐蜜及其蜜源花中的黄酮类成分

Analysis of flavonoids in acacia honey and locust flower by UPLC-MS/MS

目的:考察采用蜜源花作为"对照药材"寻找蜂蜜中特征成分的分析方法的可行性。方法:洋槐蜜和洋槐花酸水提取液经D-101型大孔吸附树脂提取纯化后采用UPLC-MS/MS分析。色谱柱采用Inertsil ODS-3柱(2.1 mm×75 mm,2μm),甲醇-0.02%甲酸为流动相梯度洗脱,流速为0.4 mL·min^(-1),柱温40℃,进样量2μL;以儿茶素、表儿茶素、芦丁、芸香柚皮苷、桑黄素、杨梅素、山柰酚、生松素等16种黄酮类对照品作为对照,采用电喷雾离子源(ESI),多反应监测(MRM)进行分析。结果:洋槐蜜与洋槐花中同时存在柚皮素、槲皮素、染料木素、木犀草素、山柰酚、芹菜素和汉黄芩素,且它们的含量在总共有组分含量中的占比呈一定比例,分别为5.75∶23.21∶5.00∶30.37∶32.47∶2.94∶0.25和2.45∶23.05∶0.88∶5.41∶66.04∶2.13∶0.03。洋槐蜜和洋槐花中的黄酮类组分及含量存在一定程度的相关性:洋槐花中含量高的化合物,洋槐蜜中也相对较高;相反则相对较低。最后,发现洋槐蜜中的潜在标志物染料木素。结论:采用蜜源花作为"对照"寻找蜂蜜中特征成分的分析方法具有可行性。

Objective： To investigate the feasibility of using nectar flower as a contrast to find chemical markers for honey. Methods： Acacia honey and aqueous hydrochloric acid extract of acacia flower were extracted and purified by D-101 macroporous resin and then analyzed by UPLC-MS/MS. Separation was performed on an Inertsil ODS-3 column （ 2.1 mm × 75 mm, 2 μm ）, with methanol-0.02% formic acid solution as mobile phase in gradient elution at a flow rate of 0.4 mL· min-1. The column temperature was controlled at 40 ℃ and the injection volume was 2 μL. Electrospray ionization （ ESI ） source was applied in multiple reaction monitoring （ MRM ） mode, taking 16 kinds of flavonoids as the reference, such as catechin, epicatechin, rutin, narirutin, morin, myricetin, kaempferol, pinocembrin, etc. Results： Naringenin, quercetin, genistein, luteolin, kaempferol, apigenin and wogonin were found in both acacia honey and acacia flower, and the proportion of each component in the total content of common components was 5.75 ： 23.21 ： 5.00 ： 30.37 ： 32.47 ： 2.94 ： 0.25 and 2.45 ： 23.05 ： 0.88 ： 5.41 ： 66.04 ： 2.13 ： 0.03, respectively. There was a certain degree of correlation between content of flavonoids in acacia honey and locust flower. Compounds with high content in acacia flower were also relatively high in acacia honey, vice versa. Finally, a potential marker of genistein was found in acacia honey. Conclusion： It was feasible to find markers for honey by using nectar flower as a ＂contrast＂.