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
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摘要
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。
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 100-1 000). 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 and10 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 artepillin 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.
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 100-1 000). 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 and10 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 artepillin 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.
引文
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[2] Zhang C P,Hu F L. Natural Product Research and Developm ent,2009,21(6):1084张翠平,胡福良.天然产物研究与开发,2009,21(6):1084
[3] Zhang C P,Wang K,Hu F L. Chinese Journal of Modern Applied Pharm acy,2013,30(1):102张翠平,王凯,胡福良.中国现代应用药学,2013,30(1):102
[4] Zhang C P,Si J J,Hong Q H,et al. Journal of Bee,2018,38(1):5张翠萍,思娟娟,洪奇华,等.蜜蜂杂志,2018,38(1):5
[5] Luo Z M.[MS Dissertation]. Beijing:Chinese Academy of Agricultural Sciences,2013罗照明.[硕士学位论文].北京:中国农业科学院,2013
[6] Gao L M,Xu X,Gao P B,et al. Journal of Chinese Institute o f Fo o d Science and Techno lo gy,2018,18(3):221高丽苗,徐响,高蓬勃,等.中国食品学报,2018,18(3):221
[7] Banskota A H,Tezuka Y,Kadota S. Phytother Res,2001,15(7):561
[8] Khalil M L. Asian Pac J Cancer Prev,2006,7(1):22
[9] Kalogeropoulos N,Konteles S J,Troullidou E,et al. Food Chem,2009,116(2):452
[10] Gao P B,Xu X,Gao L M,et al. Journal of Food Safety and Quality,2015,6(8):3172高蓬勃,徐响,高丽苗,等.食品安全质量检测学报,2015,6(8):3172
[11] Zhou M Y,Xu R H,Huang W,et al. Apiculture of China,2010,61(3):5周梦遥,徐瑞晗,黄微,等.中国蜂业,2010,61(3):5
[12] Zhang C P,Ping S,Huang S,et al. Chinese Pharmaceutical Jo urnal,2013,48(22):1889张翠平,平舜,黄帅,等.中国药学杂志,2013,48(22):1889
[13] Wang Y H,Fu C L,Zhang L,et al. Food Research and Develo pm ent,2016,37(21):1王月华,付崇罗,张丽,等.食品研究与开发,2016,37(21):1
[14] Zhang T T.[MS Dissertation]. Nanchang:Nanchang University,2017张婷婷.[硕士学位论文].南昌:南昌大学,2017
[15] Chen B.[MS Dissertation]. Nanchang:Nanchang University,2010陈滨.[硕士学位论文].南昌:南昌大学,2010
[16] Wang Y H,Chang H S,Yin X S,et al. Natural Product Research and Develo pm ent,2017,29(6):924王月华,常化松,尹旭升,等.天然产物研究与开发,2017,29(6):924
[17] Cao X P,Hu F L. Apiculture of China,2015,66(10):46曹雪萍,胡福良.中国蜂业,2015,66(10):46
[18] Cao W,Fu J F,Suo Z R,et al. Food and Fermentation Industries,2007,33(7):162曹炜,符军放,索志荣,等.食品与发酵工业,2007,33(7):162
[19] Zhang C P,Huang S,Wei W T,et al. J Food Sci,2014,79(7):1315
[20] Zhang C P,Zheng H Q,Liu G,et al. Food Chem,2011,127(1):345
[21] Wang K,Zhang C P,Hu F L. Natural Product Research and Develo pm ent,2013,25(1):140王凯,张翠平,胡福良.天然产物研究与开发,2013,25(1):140
[22] Kumazaki M,Shinohara H,Taniguchi K,et al. Phytomedicine,2014,21(8/9):1070
[23] Shen X G,Zhang C P,Hu F L. Natural Product Research and Develo pm ent,2015,27(5):915申小阁,张翠平,胡福良.天然产物研究与开发,2015,27(5):915
[24] Wang C L. Food Research and Development,2016,37(1):39王春玲.食品研究与开发,2016,37(1):39
[25] Zhou P,Shao Q Y,Xu Q H,et al. Journal of Bee,2013,33(4):4周萍,邵巧云,徐权华,等.蜜蜂杂志,2013,33(4):4
[26] GB/T 19427-2003
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