多反应监测联用Box-Behnken响应面法测定虎杖多成分含量

Simultaneous determination of multi-constituents in Polygonum cuspidatum using multiple reaction monitoring coupled with Box-Behnken response surface methodology

关键词:  虎杖  单因素优化  Box-Behnken响应面法  多反应监测  超高效液相色谱-串联质谱联用技术  含量测定

Key words:  Polygonum cuspidatum  single factor optimization  Box-Behnken response surface methodology  multiple reaction monitoring  ＵHＰLC-MS/MS  content determination

摘要:

[目的] 应用多反应监测模式超高效液相色谱-串联质谱联用技术（UHPLC-MS/MS）结合Box-Behnken响应面法实现不同批次虎杖中7种活性成分含量的准确测定。[方法] 采用单因素优化和Box-Behnken响应面法优化虎杖的超声提取条件，建立基于多反应监测模式UHPLC-MS/MS的虎杖中7种成分（虎杖苷、表儿茶素没食子酸酯、决明酮-8-O-葡萄糖苷、白藜芦醇、大黄素-1-O-葡萄糖苷、大黄素-8-O-葡萄糖苷、大黄素）同时测定方法，对17个批次不同产地虎杖药材进行质量评价。[结果] 最终获得优化的提取条件为72%甲醇，料液比1∶150，超声提取30 min。在该提取条件下，17个批次虎杖中的7种化学成分均能被准确定量，方法学考察结果显示不同化合物在其浓度范围内与峰面积呈现良好的线性关系（r²≥0.995），加样回收率均在96.2%~108%，日间精密度RSD≤3.12%，日内精密度RSD≤3.01%。含量测定结果表明17个批次不同产地虎杖药材中7种化合物的含量之间存在明显差异，其中大黄素、虎杖苷和大黄素-8-O-葡萄糖苷含量较高。[结论] 所建立的UHPLC-MS/MS检测技术可以用于虎杖多种成分的准确定量，该方法高效可靠、准确度高、稳定性好，能够为虎杖质量评价体系的构建提供参考。

Abstract:

[Objective] To achieve accurate quantification of seven active components in different batches of Polygonum cuspidatum by employing ultra-performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS) with multiple reaction monitoring(MRM) mode coupled with Box-Behnken response surface methodology. [Methods] The ultrasonic extraction conditions for Polygonum cuspidatum were optimized using single-factor experiments and Box-Behnken response surface design. A simultaneous quantification method for seven components(polydatin,epicatechin gallate,torachrysone-8-O-glucoside,resveratrol,emodin-1-O-glucoside,emodin-8-O-glucoside, and emodin) in Polygonum cuspidatum was established using UHPLC-MS/MS with MRM mode. This method was applied for quality assessment of 17 batches of Polygonum cuspidatum from different geographical origins. [Results] The optimized extraction conditions were determined as 72% methanol,a solid-to-liquid ratio of 1:150,and ultrasonic extraction for 30 min. Under these conditions,all seven chemical components in the 17 batches were accurately quantified. Methodological validation demonstrated good linearity for all compounds within their respective concentration ranges(r² ≥ 0.995),with recovery rates ranging from 96.2% to 108.2%. The relative standard deviations(RSD) for inter-day and intra-day precision were ≤ 3.12% and ≤ 3.01%,respectively. Quantitative analysis revealed significant variations in the contents of the seven compounds among the 17 batches,with emodin,polydatin,and emodin-8-O-glucoside being the most abundant. [Conclusion] The established UHPLC-MS/MS method proves to be efficient,reliable,accurate,and stable for the simultaneous quantification of multiple components in Polygonum cuspidatum,providing a valuable reference for constructing a comprehensive quality evaluation system for this herb.