基于顶空-固相微萃取-气相色谱-三重四极杆质谱联用的诃子挥发性成分分析

Analysis of volatile compounds in Chebulae Fructus based on head space-solid phase microextraction-gas chromatography-triple quadrupole mass spectrometry

目的建立一种基于顶空-固相微萃取-气相色谱-三重四极杆质谱联用技术的诃子药材中挥发性成分快速筛查与定量分析方法,并用于评价不同产地的诃子质量差异性。方法采用InertCap Pure-WAX毛细管柱(30 m×0.25 mm×0.25μm)为色谱柱;进样口温度为250℃,分流比为5∶1,高压进样压力为83.5kPa;载气为高纯氦气,载气控制方式为恒压力模式;色谱柱流量为1.43 mL/min,线速度为43.3 cm/s,吹扫流量为3.0 mL/min;程序升温:初始温度为50℃,保持5 min,以10℃/min升温至250℃,保持10 min;柱平衡时间为2.0 min。离子源为电子轰击源(EI),检测器电压0.3 kV,离子源温度为200℃,质谱传输接口温度为250℃,碰撞气为氩气;质谱监测模式为多反应监测。结果分析结果显示,在诃子药材中共筛查出86种挥发性化合物,按化合物结构可分为酯、酮、醛、烯、酚、酸、醇、苯、醚、吡嗪、酰胺11类。测定结果表明,不同产地的诃子在挥发性成分在种类上没有差异,但是不同产地的诃子中部分挥发性成分的含量存在较大差异。主成分分析结果显示,以诃子中86种挥发性成分的含量为降维指标分析,能在一定程度上区分不同产地的诃子,但区分不明显,这初步提示不同产地的诃子中挥发性成分的累积具有一定地域特征。结论建立的诃子中86种挥发性成分的快速筛查和定量分析方法,具有高通量、准确和简便的特点,对其它中药材的挥发性成分分析具有参考价值。

Objective To establish a method for rapid screening and quantitative analysis of volatile compounds in chebulae fructus by head space-solid phase microextraction-gas chromatography-triple quadrupole mass spectrometry,and this new analytical method was used to evaluate the quality difference of chebulae fructus collected in different pieces. Methods An InertCap Pure-WAX column was used as a separation column,and the detailed parameter information was 30 m × 0.25 mm × 0.25 μm. The injectin port temperature was 250 ℃. The injection mode was split injection with a ratio of 5∶1. The high pressure injection value was set to 83.5 kPa. The carrier gas was high purity helium and control mode was set to constant pressure. The column flow rate was 1.43 mL/min,the linear velocity was 43.3 cm/s,and the purge flow rate was 3.0 mL/min. The column temperature program as follows: maintain the initial temperature at 50 oC for 5 min,and increased the temperature at a rate of 10 oC per minute to 250 oC,hold for 10 min. the column equilibration time was 2 min. Mass spectrometry detector was triple-quadrupole mass spectrometer,ionization source was electron ionization,the detector voltage was 0.3 kV,and the ionization source temperature was 200 oC,collision gas was high purity argon;MS transfer line temperature was 250 oC. The monitoring mode was set to multiple reaction monitoring. Results The analysis results showed that a total of 86 kinds of volatile compounds were screened out in chebulae fructus,which can be divided into 11 types according to the compound structure,including: esters,ketones,aldehydes,alkenes,phenols,acids,alcohols,benzenes,ethers,pyrazines,amides,etc.,and this proved that the "sour gas" emitted by Chebulae Fructus was a complex odor composed of at least 86 volatile compounds. The results showed that there was not much difference in the types of volatile compounds in Chebulae Fructus in different regions,but there are large differences in the contents of some volatile compounds in Chebulae Fructus produced in different regions. The results of principal component analysis showed that the content of 86 volatile compounds in Chebulae Fructus as the dimensionality reduction index analysis could prove to a certain extent that there are differences in the quality of Chebulae Fructus collected in different pieces,but this evidence was not obvious. This result preliminarily indicated that the accumulation of volatile compounds in Chebulae Fructus had regional characteristics. Conclusion In summary,this newly developed method for rapid screening and quantitative analysis of 86 kinds of volatile compounds in Chebulae Fructus has the characteristics of high throughput,accuracy,and simplicity,and has reference value for volatile compounds analysis of other traditional Chinese medicines.