三脂清中7种成分的含量测定及醇提工艺优化

Determination of 7 Components in Sanzhiqing and Optimization of Its Ethanol Extraction Technology

关键词:  三脂清  高效液相色谱-质谱联用  多成分测定  正交试验  工艺优化

Key words:  Sanzhiqing  HPLC-MS/MS  multi-component determination  orthogonal test  technology optimization

摘要:

目的 采用HPLC-MS/MS同时测定蒙药三脂清中胡椒碱、荜茇宁、柯里拉京、没食子酸、鞣花酸、槲皮素、芦丁的含量，并优化三脂清的醇提工艺。方法 采用Shim-pack GIST-HP C₁₈色谱柱（2.1 mm×100 mm，3 mm），以0.1%甲酸水溶液-甲醇为流动相，梯度洗脱，流速为0.25 mL·min^-1，进样量2 μL；质谱采用电喷雾离子源，正、负离子扫描切换，多重反应监测模式进行定量分析。采用L₉（3⁴）正交试验设计，以上述7种成分含量的综合评分为考察指标，以料液比、乙醇浓度、提取时间、提取次数为考察因素进行三脂清醇提工艺优化。结果 三脂清中7种成分的峰面积和质量浓度在一定范围内均呈现良好的线性关系（r ≥ 0.999 1），仪器精密度、重复性、稳定性试验的RSD均<5%；加样回收率为99.54%~105.32%，RSD为1.39%~4.33%。最佳醇提工艺条件为料液比1：8，乙醇浓度40%，提取时间0.5 h，提取3次。3次放大试验验证结果显示，上述7种成分的平均含量分别为9.852，0.587，16.247，3.657，8.663，0.119，0.104 mg·g^-1。结论 建立的方法简便、灵敏、高效，可用于三脂清中多种主要活性成分的含量测定；优化所得的醇提取工艺稳定、可行。

Abstract:

OBJECTIVE To develop a method for simultaneous determination of piperine, piperlonguminine, corilagin, gallic acid, ellagic acid, quercetin, rutin in Mongolian medicine Sanzhiqing by HPLC-MS/MS, and to optimize the ethanol extraction technology. METHODS The separation was performed on Shim-pack GIST-HP C₁₈ column(2.1 mm×100 mm, 3 μm), mobile phase consisting of water(containing 0.1% formic acid) and methanol with gradient elution at a flow rate of 0.25 mL·min^-1, and the injection volume was 2 μL. Multiple-reaction monitoring was employed with switching electrospray ion source polarity in positive and negative mode. With comprehensive score of the content of 7 ingredients as the inspection index and material-liquid ratio, ethanol concentration, extraction time and extraction times as the inspection factor, the L₉(3⁴) orthogonal test design was used to optimize the ethanol extraction technology of Sanzhiqing. RESULTS There was a good linear relationship between the peak area and the mass concentration of the 7 components in Sanzhiqing(r ≥ 0.999 1). The RSD of instrument precision, repeatability and stability were all <5%, and the average recoveries ranged from 99.54% to 105.32% and the RSD ranged from 1.39% to 4.33%. The optimal ethanol extraction technology included solid-liquid ratio of 1:8, ethanol concentration of 40%, extracting for 0.5 h and extracting for 3 times. Results of three validation tests showed that average content of the above 7 ingredients was 9.852, 0.587, 16.247, 3.657, 8.663, 0.119, 0.104 mg·g^-1 respectively. CONCLUSION The established method is simple, sensitive and efficient, and can be used for determine the content of various main active ingredients in Sanzhiqing, and the optimized ethanol extraction technology is stable and feasible.