超高效液相色谱-串联质谱法检测 小菜蛾血清中的β-蜕皮激素
A method for using UPLC-MS/MS to measure β-ecdysone in Plutella xylostella
孙诗晴；王念猛；薛超彬
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DOI：10.7679/j.issn.2095-1353.2020.052
作者单位：山东农业大学植物保护学院，农药毒理与应用技术省级重点实验室，泰安 271018；山东农业大学植物保护学院，农药毒理与应用技术省级重点实验室，泰安 271018；山东农业大学植物保护学院，农药毒理与应用技术省级重点实验室，泰安 271018
中文关键词：β-蜕皮激素；小菜蛾；芸苔素内酯；内标法；定量限；高效液相色谱-串联质谱
英文关键词：β-ecdysone; Plutella xylostella; brassinolide; internal standard method; limit of quantitation; UPLC-MS/MS
中文摘要：
【目的】 研究建立可以准确检测小菜蛾血清中痕量β-蜕皮激素的超效液相色谱-串联质谱（UPLC-MS/MS）分析法。【方法】 小菜蛾血清β-蜕皮激素样品用乙腈提取并进行蛋白沉淀，以C18色谱柱分离，经UPLC-MS/MS检测，以芸苔素内酯为内标物，内标法定量。【结果】 结果表明，在0.5-50 μg/L浓度范围内β-蜕皮激素线性相关系数R2=0.999 8；在0.5、10和50 µg/L 3个添加水平下的平均回收率在91.9%-104.9%之间，相对标准偏差（RSDS，n=5）在3.2%-9.2%之间；方法的定量限为0.5 µg/L。【结论】 该方法具有操作简单、灵敏度高，稳定性好，应用性强等优点，可为昆虫血清中的β-蜕皮激素检测提供科学准确的方法。
英文摘要：
[Objectives] A method for using UPLC-MS/MS to detect β-ecdysone
in the serum of Plutella xylostella was developed. [Methods] Serum β-ecdysone
of P. xylostella was extracted with
acetonitrile and precipitated with protein. It was then separated in a C18 column, analyzed with UPLC-MS/MS and the β-ecdysone
content determined using the internal standard brassinolide. [Results] The results show that the linear correlation coefficient of β-ecdysone was R2 = 0.999 8 in the range of 0.5-50 μg/L, the average recovery was
91.9%-104.9% at levels of 0.5 μg/L,
10 μg/L and 50 μg/L, the relative standard deviation
(RSDs, n = 5) was 3.2%-9.2% and the quantitative limit of the method was
0.5 μg/L. [Conclusion] This method
has the advantages of being relatively simple, sensitive, stable and
applicable. It should provide an accurate method for the detection and
measurement of β-ecdysone in the
serum of other insects.