蜂蜜炼制前后黄酮类成分种类和含量变化分析

Comparative analysis of species and contents of flavonoids before and after honey refining

目的研究中药辅料蜂蜜炼制前后黄酮类成分变化。方法参考《中药药剂学》,对不同来源的6种蜂蜜进行炼制。供试品经固相萃取法富集黄酮类成分。采用高效液相色谱-三重四级杆串联质谱(HPLC-TQ-MS)法测定,Thermo Accucore RP-MS色谱柱(100 mm×2.1 mm,2.6μm),以0.1%甲酸水溶液-甲醇为流动相进行梯度洗脱,体积流量0.3 m L/min,柱温30℃。质谱条件:采用电喷雾离子源(ESI),正离子检测模式,多反应监测模式(MRM)扫描分析。结果蜂蜜中共检测到12种黄酮类成分,包括黄酮苷元、二氢黄酮苷元和异黄酮苷元。黄酮苷元有8种:槲皮素、桑黄素、木犀草素、山柰酚、芹菜素、汉黄芩素、高良姜素和白杨素;二氢黄酮苷元有3种:短叶松素、柚皮素和乔松素;异黄酮苷元染料木素。不同蜜源蜂蜜所含黄酮类成分种类和含量存在明显差异。6个蜂蜜样品炼制后,都发现了蜂蜜中没有报道过的芦丁和芸香柚皮苷,12种检测到的蜂蜜黄酮类成分在炼制后其量都有不同程度的变化,椴树蜜的槲皮素、桑黄素、木犀草素、山柰酚、芹菜素、汉黄芩素、乔松素和白杨素量均有增加,短叶松素、柚皮素和高良姜素量没增加反而还有下降的;洋槐蜜中除芹菜素量增加不明显外,其余均有所增加,尤其是槲皮素和桑黄素增加的较为明显;百花蜜和枣花蜜12种黄酮类成分量均显著增加。结论炼制可造成蜂蜜中黄酮类成分的种类和含量发生改变。

Objective To study the changes of flavonoid components in honey before and after refining. Methods Six batches of honey from different sources were collected and refined based on Chinese Drugs Pharmacy. Solid phase extraction was used to enrich flavonoids from samples. HPLC-TQ-MS was established to determine the contents of 15 flavonoids in crude and refined honey, the separation was performed on a Thermo Accucore RP-MS （100 mm×2.1 mm, 2.6 μm） column with the gradient elution of methanol-0.1% formic acid water, the flow rate was 0.3 mL/min, and the column temperature was 30℃. MS condition： Electrospray ionization （ESI） source was applied and operated in the positive multiple reaction monitoring （MRM） modes. Results Twelve kinds of flavonoids in three species, flavonoids, dihydrofiavonoids, and isoflavones, were detected in honey, which including quercetin, morin, xanthophyllin, kaempferol, apigenin, wogonin, galangin, chrysin in flavonoids; pinobanksin, naringin, and pinocembrin in dihydroflavonoids; genistein in isoflavones. There was significant difference in the species and contents of flavonoids between crude honey and refined honey. After refining, rutin and narirutin were detected which have not been reported in references, and the contents of 12 kinds of flavonoids have increased at different degrees. The contents of quercetin, morin, xanthophyllin, kaempferol, apigenin, wogonin, pinocembrin, and chrysin in linden honey have increased, but the contents of pinobanksin, naringin, and galangin have decreased; all the contents of the components in acacia honey have increased except apigenin, especially the quercetin and morin; The content of 12 kinds offlavonoids in the honey of various flowers and Chinese date honey all have increased. Conclusion Refining can change the species and contents of flavonoids in honey.