超高效液相色谱-串联三重四极杆质谱法快速测定食品中巴豆苷、木兰花碱和异鸟嘌呤

Rapid determination of crotonoside, magnoflorine and isoguanine in foods by ultra performance liquid chromatography-tandem mass spectrometry

中文关键词:  巴豆苷  木兰花碱  异鸟嘌呤  UPLC-MS/MS

英文关键词:crotonoside  magnoflorine  isoguanine  ultra performance liquid chromatography-tandem mass spectrometry

中文摘要:

      目的 建立超高效液相色谱-串联三重四极杆质谱法(ultra performance liquid chromatography-tandem mass spectrometry, UPLC-MS/MS)快速测定食品中巴豆苷、木兰花碱和异鸟嘌呤的分析方法。方法 样品采用60%(体积分数)乙腈水溶液提取, 提取液经涡旋振荡、超声提取、高速离心后吸取上清液经滤膜过滤后用于测定。以乙腈-0.1%(体积分数)甲酸水溶液为流动相梯度洗脱, 经Waters BEH Amide (3.0 mm×100 mm, 2.5 μm)色谱柱分离, 质谱采用正离子电喷雾离子化(positive ion electrospray ionization, ESI+)、多反应监测(multiple reaction monitoring, MRM)模式对巴豆苷、木兰花碱和异鸟嘌呤3种生物碱进行定性和定量测定。结果 3种生物碱在对应质量浓度范围内线性关系良好(r>0.997)平均加标回收率范围为83.7%~102.3%, 相对标准偏差为3.2%~7.4% (n=6)。巴豆苷、木兰花碱的检出限均为0.1 μg/L, 定量限均为1.0 μg/L; 异鸟嘌呤的检出限为1.0 μg/L, 定量限为2.0 μg/L。结论 本方法样品前处理快速简便, 耗时短, 方法灵敏度高, 准确性好, 适用于疑似巴豆中毒应急事件中3种生物碱的快速检测与确证分析。

英文摘要:

      Objective To establish a rapid determination and analysis method for crotonoside, magnoflorine and isoguanine in food by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Methods The samples were extracted with 60% (volume fraction) acetonitrile aqueous solution. After vortex oscillation, ultrasonic extraction and high-speed centrifugation, the supernatant was filtered through a filter membrane and used for determination. The mobile phase consisted of acetonitrile-0.1% (volume fraction) formic acid aqueous solution for gradient elution. Separation was carried out on a Waters BEH Amide (3.0 mm×100 mm, 2.5 μm) chromatography column. The mass spectrometry was conducted using positive ion electrospray ionization (ESI+) and multiple reaction monitoring (MRM) mode for qualitative and quantitative determination of 3 kinds of alkaloids, namely crotonoside, magnoflorine and isoguanine. Results The linear relationship between 3 kinds of alkaloids was good within the corresponding mass concentration range (r>0.997), the average recoveries ranged from 83.7% to 102.3%, and the relative standard deviations were ranged from 3.2% to 7.4% (n=6). The limits of detection for crotonoside and magnoflorine were both 0.1 μg/L, and the limits of quantification were 1.0 μg/L. The limit of detection for isoguanine was 1.0 μg/L, and the limit of quantification was 2.0 μg/L. Conclusion This method has a rapid and simple sample pretreatment process, short time-consuming, high sensitivity and good accuracy, and is suitable for the rapid detection and confirmatory analysis of 3 kinds of alkaloids in emergency incidents of suspected croton poisoning.