目的:建立基于QuEChERS结合超高效液相色谱-串联质谱(UPLC-MS/MS)测定蜂蜜中28个吡咯里西啶生物碱的快速检测方法,并根据测定结果进行初步的风险评估。方法:采用QuEChERS法对样品进行前处理,ACQUITY UPLC HSS T3色谱柱(2.1 mm×100 mm, 1.8 μm),以含0.05%甲酸和2.5 mmol·L-1甲酸铵的水溶液(A)及含0.05%甲酸和2.5 mmol·L-1甲酸铵的乙腈溶液(B)为流动相,梯度洗脱,流速为0.3 mL·min-1,柱温为40 ℃;电喷雾离子源为正离子多反应监测模式,外标法测定。结果:28个吡咯里西啶生物碱在1~20 ng·mL-1内线性关系良好,相关系数均大于0.99;平均回收率为71.8%~103.5%,RSD为3.3%~9.2%。28个吡咯里西啶生物碱的定量下限为0.20~0.90 ng·mL-1;30批样品中有10批样品检出吡咯里西啶生物碱,阳性样品中总含量为4.85~78.34 μg·kg-1。结论:该方法操作简便快速、准确、灵敏,可同时测定蜂蜜中28个吡咯里西啶生物碱含量。初步风险评估显示,蜂蜜存在一定的安全风险。
昝珂, 李耀磊, 王莹, 刘丽娜, 金红宇, 左甜甜, 马双成
. QuEChERS-UPLC-MS/MS法快速测定蜂蜜中28个吡咯里西啶生物碱的含量及风险评估*[J]. 药物分析杂志, 2021
, 41(12)
: 2087
-2094
.
DOI: 10.16155/j.0254-1793.2021.12.06
Objective: To establish a method for the simultaneous determination of 28 pyrrolizidine alkaloids in honey by QuEChERS and UPLC-MS/MS, and carry out preliminary risk assessment according to the research results. Methods: The samples were prepared by QuEChERS, then separated on an ACQUITY UPLC HSS T3 column (2.1 mm×100 mm, 1.8 μm). The sample was eluted with a gradient mobile phase consisting of 0.05% formic acid and 2.5 mmol·L-1 ammonium formate in water(A)-0.05% formic acid and 2.5 mmol·L-1 ammonium formate in acetonitrile(B). The flow rate was 0.3 mL·min-1 and the column temperature was 40 ℃. The MS system was operated by using electrospray ionization (ESI) in the positive ion mode, and the scan mode was in multiple reactions monitoring (MRM) mode. External standard method was used in the experiment. Results: The results showed that all the 28 PAs had good linearity in 1-20 ng·mL-1 with the correlation coefficients higher than 0.99. The average recoveries of all the 28 PAs were between 71.8% and 103.5%, with the RSDs of 3.3%-9.2%. The limits of quantifications (LOQs) of 28 PAs were between 0.20 ng·mL-1 and 0.90 ng·mL-1. PAs was detected in 10 of 30 samples, and the total contents of positive samples were 4.85-78.34 μg·kg-1. Conclusion: The established method can simultaneously determine 28 pyrrolizidine alkaloids in honey. The preliminary risk assessment shows that there is a certain risk of drug safety.
[1] 高江国, 王长虹, 李岩, 等. 吡咯里西啶生物碱的药理作用、毒性及药(毒)物代谢动力学研究进展[J].中国中药杂志, 2009, 34(5):506
GAO JG, WANG CH, LI Y, et al. Advance on pharmacologic actions, toxicity and pharmacokinetics of pyrrozidine alkaloids[J].China J Chin Mater Med, 2009, 34(5): 506
[2] STEINHOFF B. Pyrrolizidine alkaloid contamination in herbal medicinal products: limits and occurrence[J].Food Chem Toxicol, 2019, 130: 262
[3] 王涛, 宋海波. 吡咯里西啶类生物碱肝毒性研究进展及风险分析[J].医药导报, 2018, 37(9):1033
WANG T, SONG HB. Research progress and risk analysis on hepatotoxicity of pyrrolizidine alkaloids [J].Her Med, 2018, 37(9):1033
[4] SCHRENK D, GAO L, LIN G, et al. Pyrrolizidine alkaloids in food and phytomedicine: occurrence, exposure, toxicity, mechanisms, and risk assessment—A review[J].Food Chem Toxicol, 2020, 136: 1
[5] 郑亚杰,刘秀斌,林莉,等. 蜂蜜中植物源性毒性成分的研究进展[J].食品科学, 2019, 40(3): 307
ZHENG YJ, LIU XB, LIN L, et al. Plant-derived toxins in honey: a review [J].Food Sci, 2019, 40(3): 307
[6] GRIFFIN CT,DANAHER M,ELLIOTT CT, et al. Detection of pyrrolizidinealkaloids in commercial honey using liquid chromatography–ion trap mass spectrometry[J].Food Chem,2013,136(3-4):1577
[7] CRAMER L,BEUERLE T. Detection and quantification of pyrrolizidine alkaloids in antibacterial medical honeys[J].Planta Med,2012,78(18):1976
[8] KEMPF M,WITTIG M,REINHARD A, et al. Pyrrolizidine alkaloids in honey: comparison of analytical methods[J].Food Addit Contam A,2011,28(3):332
[9] WANG T,FRANDSEN HL,CHRISTIANSSON NR, et al. Pyrrolizidine alkaloids in honey: quantification with and without standards[J].Food Control,2019,98:227
[10] BEALES KA,BETTERIDGE K,COLEGATE SM, et al. Solid-phase extraction and LC-MS analysis of pyrrolizidine alkaloids in honey[J].J Agric Food Chem,2004,52(21):6664
[11] 中华全国供销合作总社. 蜂蜜中5种双稠吡咯啶类生物碱的测定. 高效液相色谱-质谱/质谱法(GH/T 1107-2015)[S].2015
All China Federation of Supply and Marketing Cooperatives. Determination of 5 pyrrolizidine alkaloids in honey by HPLC-MS/MS(GH/T 1107-2015)[S].2015
[12] European Medicines Agency. Public statement on contamination of herbal medicinal products/traditional herbal medicinal products with pyrrolizidine alkaloids [R/OL].[2016-5-31].http://www.ema.europa.eu/docs/en_gb/document_library/public_statement/2016/06/wc500208195.pdf
[13] KIM L, LEE D, CHO HK, et al. Review of the QuEChERS method for the analysis of organic pollutants: persistent organic pollutants, polycyclic aromatic hydrocarbons, and pharmaceuticals [J].Trends Environ Anal,2019,22:e00063
[14] MARTINELLO M,BORIN A,STELLA R, et al. Development and validation of a QuEChERS method coupled to liquid chromatography and high resolution mass spectrometry to determine pyrrolizidine and tropane alkaloids in honey[J].Food Chem,2017,234:295
[15] 中华人民共和国药典 2020年版.一部[S].2020:374
ChP 2020. Vol Ⅰ[S].2020:374
