基于气相色谱-质谱联用的小儿柴胡退热栓主要药效成分质量控制方法研究

Quality Control Method of Main Active Components in Xiao’er Chaihu Tuire Suppository Based on Gas Chromatography-Mass Spectrometry

目的:建立基于气相色谱-质谱(GC-MS)联用的小儿柴胡退热栓主要药效成分的质量控制方法。方法:采用GC-MS联用分析技术,测定小儿柴胡退热栓的主要药效成分正己醛和正庚醛的含量,并结合质谱数据库信息对未知成分进行比对分析;使用SH-Rtx-Wax毛细管色谱柱(30 m×0.25 mm×0.25μm)分离,程序升温条件为初始温度60℃,保持2 min,以20℃/min速率升温至140℃,保持3 min,再以15℃/min速率升温至180℃,保持4 min,最后以10℃/min速率升温至230℃,保留4 min;载气为氦气,流速为1 mL/min,分流比为10∶1;进样口温度为230℃,进样量1.0μL;质谱条件:EI正离子模式,离子源电压70 eV,离子源温度230℃,质量扫描范围为45~300,数据采集模式为Q_(3)Scan。结果:正己醛和正庚醛在选定的浓度范围内与各自峰面积呈良好的线性关系,相关系数分别为0.998和0.999,加样回收率分别为99.7%和100.4%,RSD分别为1.46%和2.89%,重复性、稳定性、精密度均良好,测得小儿柴胡退热栓中正己醛和正庚醛的平均含量分别为每枚73.3μg和110.3μg,并鉴定出包括β-桉叶醇、4-萜烯醇等26个化合物成分。结论:基于GC-MS的小儿柴胡退热栓主要药效成分的质量控制方法专属性强、灵敏度高、重复性好,可作为其质量控制方法,为小儿柴胡退热栓的进一步药理作用物质基础研究提供一定的理论基础。

Objective: To establish a quality control method for the main active components of Xiao’er Chaihu Tuire suppository based on gas chromatography-mass spectrometry(GC-MS). Methods: The contents of N-hexanal and N-heptanal in the main active components of Xiao’er Chaihu Tuire suppository were determined by GC-MS, and the unknown components were compared and analyzed combined with the mass spectrometry database. The samples were separated on a SH-Rtx-Wax(30 m×0.25 mm×0.25 μm) capillary column under the initial programmed temperature of 60 ℃ for 2 mins, the temperature was increased to 140 ℃ at the rate of 20 ℃/min for 3 mins, to 180 ℃ at the rate of 15 ℃/min for 4 mins, and finally to 230 ℃ at a rate of 10 ℃/min for 4 mins. GC-MS carrier gas was helium, with a flow rate of 1 mL/min and a split ratio of 10∶1. The inlet temperature was 230 ℃, and the injection volume was 1.0 μL. Mass spectrometry conditions were as follows: the EI source, positive ion mode, bombardment voltage at 70 eV, ion source temperature at 230 ℃, the quality scan range from 45 to 300, and data acquisition mode of Q_(3)Scan. Results: N-hexanal and N-heptanal had good linear relationship with the peak areas in the selected concentration range. The correlation coefficients were 0.998 and 0.999, the recovery rates were 99.7% and 100.4%, and RSD were 1.46% and 2.89%, respectively. The repeatability, stability and precision were all good. The average contents of N-hexanal and N-heptanal in Xiao’er Chaihu Tuire suppository were respectively 73.3 μg/piece and 110.3 μg/piece, and 26 compounds were identified, including β-eudesmol and terpinen-4-ol. Conclusion: The quality control method of the main pharmacodynamic components of Xiao’er Chaihu Tuire suppository based on GC-MS has strong specificity, high sensitivity and good repeatability, and can be used as quality control method to provide theoretical basis for further research on the material basis of the pharmacological action of Xiao’er Chaihu Tuire suppository.