正交设计结合响应曲面法研究白芥子中芥子碱的提取工艺

目的建立测定白芥子中芥子碱含量的高灵敏质谱方法,进一步优化并获得药材中芥子碱的最佳提取工艺。方法高效液相色谱-电喷雾-串联质谱法测定芥子药材中芥子碱的含量,以芥子碱含量为指标,以热醇回流为提取方法,采用正交设计和响应曲面2种方法优化白芥子药材中芥子碱的提取工艺。结果芥子碱质量浓度在15.5~294.5μg·mL~(-1)范围内提取离子峰峰面积与浓度呈良好的线性关系,方法检测限为0.1ng·mL~(-1),白芥子中芥子碱的最佳提取工艺为:40倍量体积分数为65%的乙醇,60℃加热回流提取5.0h。结论高效液相色谱-串联质谱法具有特异性强的特点,可作为白芥子中芥子碱的含量测定方法;正交设计结合响应曲面法能准确确定白芥子中芥子碱的最佳提取工艺,能为其他中药提取工艺的优化提供借鉴。

Study on the extraction procedure of sinapine in white mustard seed by orthogonal design and response surface methodology

Objective To develop a high sensitive method for the determination of sinapine in white mustard seed and to optimize the extraction procedure of sinapine .Methods The content of sinapine in white mustard seed was determined by high performance liq-uid chromatography-tandem mass spectrometry .Then the concentration of sinapine was used as an index to optimize the ethanol extraction procedure of sinapine from white mustard seed by orthogonal design and response surface methodology .Results The area of extracting peak of sinapine was linearly corresponded to its concentrations in the range of 15 .5-294 .5 μg?mL -1 .The de-tection limit of the method was 0 .1 ng?mL -1 .The optimized conditions were confirmed to be 5 h of heating reflux by forty folds of 65% ethanol at 60 ℃ .Conclusion High performance liquid chromatography-tandem mass spectrometry has the advantages of high specificity .This property enables it to become the preferable method for measuring sinapine in white mustard seed .The com-bination of orthogonal design with response surface methodology is capable of establishing the optimized extracting procedure of si-napine in w hite mustard seed .