超高效液相色谱⁃串联质谱检测枸杞中苦参碱和氧化苦参碱残留量的方法

Ultra High Performance Liquid Chromatography⁃Tandem Mass Spectrometry for Detection of Matrine and Oxymatrine Residues in Goji Berries

使用超高效液相色谱⁃串联质谱仪,采用同位素内标法定量,建立枸杞中苦参碱和氧化苦参碱的测定方法。样品经1.0%磷酸水溶液超声提取,强阳离子固相萃取柱净化,10 mL 5%氨水甲醇溶液洗脱,采用0.1%甲酸水溶液和甲醇为流动相,梯度洗脱,通过T3色谱柱分离,电喷雾离子源正离子扫描模式、多反应监测模式检测。通过考察不同种类提取溶剂、超声时间、固相萃取条件下目标化合物峰面积,确定最优前处理方式。结果表明:苦参碱和氧化苦参碱在0.5~50.0μg/kg范围内呈现良好线性关系,相关系数(R2)均大于0.999。方法检出限0.05μg/kg,定量限0.2μg/kg。低、中、高3个浓度加标回收试验,苦参碱回收率范围在76.3%~90.7%,相对标准偏差(relative standard deviation,RSD)在2.2%~7.4%,氧化苦参碱回收率范围在79.2%~89.0%,RSD在3.1%~6.6%。该方法适用于枸杞中苦参碱和氧化苦参碱检测。

A method for the determination of matrine and oxymatrine in goji berries was established by using ultra high performance liquid chromatography⁃tandem mass spectrometry and isotope internal standard method for quantification.The sample was extracted by ultrasound with a 1.0%phosphoric acid aqueous solution,puri⁃fied by a strong cation solid⁃phase extraction column,and eluted with a 5%ammonia methanol solution of 10 mL.The sample was then eluted with a 0.1%formic acid aqueous solution and methanol as the mobile phase.Gradi⁃ent elution was performed,separated by a T3 chromatographic column,and detected by electron spray ionizq⁃tion positive ion mode multiple reaction mode(MRM).The optimal pre⁃treatment method was determined by examining the response values of target compounds under different types of extraction solvents,ultrasound time,and solid⁃phase extraction conditions.The results showed that there was a good linear relationship be⁃tween matrine and oxymatrine in the range of 0.5-50.0μg/kg,with correlation coefficients(R2)greater than 0.999.The detection limit of the method was 0.05μg/kg,and the quantitative limit was 0.2μg/kg.The recov⁃ery experiment of adding standard at three concentrations of low,medium,and high showed that the recovery rate of matrine ranged from 76.3%to 90.7%,and the relative standard deviation(RSD)ranged from 2.2%to 7.4%;the recovery rate of oxymatrine ranged from 79.2%to 89.0%,and the RSD ranged from 3.1%to 6.6%.This method is fast,simple,accurate,and reproducible,and is suitable for the determination of matrine and oxymatrine in goji berries.