LC-MS/MS法同时测定刺五加制剂中紫丁香苷、刺五加苷E和异嗪皮啶的含量

Simultaneous Determination of Syringin, Eleutheroside E and Isofraxidin in Acanthopanax Preparations by LC-MS/MS

目的：建立一种液相色谱-串联四级杆质谱（LC-MS/MS）法同时测定两种刺五加制剂中紫丁香苷、刺五加苷E和异嗪皮啶的含量。方法LC采用内标法，MS采用电喷雾离子源，多反应监测模式，正负离子同时检测。3种被测成分的监测离子对分别为395.0/232.0（紫丁香苷），765.2/765.2（刺五加苷E），223.0/162.0（异嗪皮啶），内标化合物的监测离子对为431.0/311.1（牡荆苷）。色谱柱为Diamonsil C18色谱柱（4.6 mm×150 mm ,5μm），流动相A为甲醇，流动相B为0.1%甲酸溶液，梯度洗脱，流速0.6 mL/min，分析时间12 min。结果刺五加注射液、刺五加片中紫丁香苷、刺五加苷E、异嗪皮啶的线性范围分别为6.00-2000 ng/mL（r =0.9979），6.00-2000 ng/mL（r =0.9951），2.00-600 ng/mL （r=0.9931）；刺五加注射液的平均加样回收率（n=9）分别为104.13%，95.16%，102.64%；刺五加片的平均加样回收率（n=9）分别为102.51%，99.60%，99.15%。结论采用LC-MS/MS方法测定刺五加注射液、刺五加片中3个有效成分含量，该方法快速、简便、灵敏度高、专属性好，可作为刺五加制剂的一种质量控制评价方法。

Objective To establish a liquid chromatography-tandem quadrupole mass spectrometry（LC-MS/MS） method for simultaneous determination of syringin, eleutheroside E and isofraxidin in Acanthopanax preparations. Methods Vitexin was used as internal standard for the determination of three active compounds in Acanthopanax preparations. Detection and quantification were performed by mass spectrometry in multiple reaction monitoring（MRM） mode with m/z 395.0→232.0 for syringin, m/z 765.2→765.2 for eleutheroside E, 223.0→162.0 for isofraxidin, and m/z 431.0→311.1 for vitexin, respectively. The chromatographic analysis was carried out by Diamonsil C18 column（4.6 mm × 150 mm, 5 μm）. The gradient elution was performed with mobile phase consisted of methanol（A）-0.1%formic acid in water （B）, with at a flow rate of 0.6 mL/min. The acquisition time was 12 min for each run. Results The calibration curves were linear in the ranges of 6.00-2000 ng/mL（r=0.9979） for syringin,6.00-2000 ng/mL（r=0.9951） for eleutheroside E, and 2.00-600 ng/mL（r=0.9931） for isofraxidin. The average recoveries （n=9） were 104.13 %,95.16 %,102.64 % for Acanthopanax injection and 102.51%,99.60 %,99.15% for Acanthopanax tablets, respectively. Conclusion This method was successfully applied for the determination of three active ingradients in Acanthopanax preparations and the method was rapid, simple, sensitive and specific, which could become an effective method for the quality control of Acanthopanax preparations.