高效液相色谱-四极杆飞行时间质谱检测中国杨树型蜂胶、巴西绿蜂胶和杨树胶中的酚类化合物及真伪鉴别

建立了用于检测中国杨树型蜂胶、巴西绿蜂胶和杨树胶中23种酚类化合物的高效液相色谱-四极杆飞行时间质谱(HPLC/Q-TOF MS)法,并进行了差异性分析。蜂胶和树胶样品用甲醇-水(1∶1,v/v)溶解后,过0. 45μm有机相滤膜后进样。采用Agilent Eclipse Plus C18色谱柱分离,以乙腈和0. 1%(体积分数)甲酸水溶液为流动相进行梯度洗脱,电喷雾正离子模式全扫描方式检测,扫描范围为m/z 100～1 000,外标法定量。结果表明,23种酚类化合物在10～200μg/L范围内线性关系良好,相关系数均大于0. 99。桔皮素和刺芒柄花素的检出限和定量限分别为0. 2和1μg/L,其他化合物的检出限和定量限分别为2和10μg/L。在10、25和50 m g/kg 3个添加水平下,23种酚类化合物在3种样品基质中的提取回收率为70. 2%～122. 6%,RSD值均小于10%。水杨苷、肉桂酸、咖啡酸和香豆酸可以作为中国杨树型蜂胶掺假鉴别的特征化合物,咖啡酸、阿魏酸、白杨素、咖啡酸苯乙酯、松属素、高良姜素、香豆酸、异鼠李素、山柰素和阿替匹林C可以作为辨别中国杨树型蜂胶和巴西绿蜂胶的特征化合物。该结果对蜂胶产品的质量控制具有一定的参考意义。

Determination of phenolic compounds in Chinese poplar propolis,Brazil green propolis,and poplar gum by high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry and preliminary study of the identification of adulteration

High-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (HPLC/Q-TOF MS) was used to analyze 23 phenolic compounds in Chinese poplar propolis, Brazil green propolis, and poplar gum. Propolis and poplar gum samples were dissolved in methanol-water (1:1, v/v) and filtered by 0.45 μm organic phase filtration membrane. The separation was carried out in an Agilent Eclipse Plus C18 column with gradient elution using acetoni-trile and 0.1% (volume percentage) formic acid solution as the mobile phases. The compounds were detected using positive ion electrospray ionization in full scan mode (m/z 100-1000). The quantification analysis was performed by the external standard method. The results showed that all the compounds were linear, with correlation coefficients>0.99, in the range of 10-20 μg/L. The limits of detection and limits of quantification for tangeretin and formononetin were 0.2 and 1 μg/L, respectively, and those for the other compounds were 2 and 10 μg/L, respectively. At the spiked levels of 10, 25, and 50 mg/kg, the recoveries of all the compounds ranged from 70.2% to 122.6%, with relative standard deviations of less than 10%. Salicin, cinnamic acid, caffeic acid, and coumaric acid could be used as markers for detecting adulteration in Chinese poplar propolis, while caffeic acid, ferulic acid, chrysin, caffeic acid phenethyl ester, pinocembrin, galangin, coumaric acid, isorhamnetin, kaempferide, and arte-pillin C could be used as markers for identifying Chinese poplar propolis and Brazil green propolis. The results presented in this paper may be helpful in quality control of propolis products.