高效液相色谱-串联质谱法测定芝麻菜中萝卜硫素

Determination of sulforaphane in Eruca sativa Mill by high performance liquid chromatography-tandem mass spectrometry

中文关键词:  萝卜硫素  芝麻菜  高效液相色谱-串联质谱法

英文关键词:sulforaphane  Eruca sativa Mill  high performance liquid chromatography-tandem mass spectrometry

中文摘要:

      目的 建立高效液相色谱-串联质谱法测定芝麻菜中萝卜硫素的分析方法。方法 芝麻菜样品加入一定量pH 6磷酸缓冲溶液, 于50 ℃水浴下振荡水解2 h, 冷却至室温后经二氯甲烷萃取, 萃取液吹干后用乙腈复溶, 以0.1%甲酸水溶液-乙腈为流动相进行梯度洗脱, 经Zorbax SB C18色谱柱(100 mm×2.1 mm, 3.5 μm)进行分离, 采用电喷雾正离子多反应监测(multiple response monitoring, MRM)模式检测, 外标法定量测定新鲜芝麻菜中萝卜硫素的含量。结果 萝卜硫素在10~500 ng/mL质量浓度范围内线性关系良好(r2≥0.999), 检出限为4.0 μg/kg, 定量限为13.0 μg/kg, 在低、中、高3水平的加标回收率为95.7%~104.0%, 相对标准偏差低于1.000%。应用本研究建立的方法对不同生长周期新鲜芝麻菜中萝卜硫素含量检测, 变化趋势与文献报道结果一致, 萝卜硫素在生长过程中逐渐积累, 成熟前期达到最高, 到成熟后期由于植物衰老萝卜硫素含量存在下降趋势。结论 本方法满足准确度、精密度及灵敏度等要求, 适用于新鲜芝麻菜中萝卜硫素的检测。

英文摘要:

      Objective To establish a method for the determination of sulforaphane in Eruca sativa Mill by high performance liquid chromatography-tandem mass spectrometry. Methods The samples were added to pH 6 phosphate buffer solution and hydrolyzed by shaking in 50 ℃ water bath for 2 h. After cooling to room temperature and extraction by dichloromethane, the extract was blown dry and re-solubilized with acetonitrile, separated on Zorbax SB C18 column (100 mm×2.1 mm, 3.5 μm) using 0.1% formic acid aqueous solution and acetonitrile as mobile phase, glucoerucin in fresh Eruca sativa Mill was detected by electrospray positive ion and multiple response monitoring (MRM) mode, and quantified by external standard method. Results The linear range of sulforaphane was 10?500 ng/mL (r2≥0.999), the limit of detection was 4.0 μg/kg, and the limit of quantification was 13.0 μg/kg. The spiked recoveries ranged from 95.7% to 104.0% at the low, medium and high levels, and the relative standard deviations were less than 1.0%. The method established in this study was applied to determine the content of sulforaphane in fresh Eruca sativa Mill of different growth cycles, and the change of trend were basically consistent with the literature reports. Sulforaphane accumulated gradually accumulated during the growth and reached a maximum at the early maturity. However, the sulforaphane content had a decreasing trend at the late maturity due to plant senescence. Conclusion This method meets the requirements of accuracy, precision and sensitivity, and can be used for the determination of sulforaphane in Eruca sativa Mill.