目的 采用超高效液相色谱串联三重四级杆质谱法（UPLC-QqQ-MS/MS）技术建立了同时测定热毒宁注射液中6个活性成分[japonicaside A（JA）、L-phenylalanion secologanin B（PSB）、木犀草苷、东莨菪内酯、（1S，6R，7R，10R）-6-羧基-10-甲基-α-亚甲基-1-（1-氧丁基）-环己烷丙烯酸（CMCA）、3α，5α-tetrahydrodeoxycordifoline lactam（TL）]的定性定量的检测方法。方法 色谱条件为Agilent Zorbax SB-Aq C₁₈色谱柱（150 mm×2.1 mm，3.5 μm），流动相为0.1%甲酸水溶液-甲醇，梯度洗脱：0.01~2.00 min，5%甲醇；2.00~4.00 min，5%~40%甲醇；4.00~11.00 min，40%~95%甲醇；11.00~13.00 min，95%甲醇；13.00~13.10 min，95%~5%甲醇；13.10~14.00 min，5%甲醇；体积流量0.5 mL/min；柱温20℃。质谱条件：采用电喷雾电离源（ESI），扫描方式为正、负离子多反应监测模式。其中，正离子监测模式的质谱条件为碰撞气（CAD）体积流量8 mL/min，气帘气（CUR）体积流量20 mL/min，雾化温度（TEM）500℃，喷雾电压（IS）4 500 V；负离子监测模式的质谱条件为碰撞气（CAD）体积流量8 mL/min，气帘气（CUR）体积流量20 mL/min，TEM 500℃，喷雾电压（IS）-4 500 V。结果 热毒宁注射液中6个活性成分在测定的质量浓度范围内均具有良好的线性关系（r ≥ 0.999 0），平均加样回收率分别为78.93%、114.65%、101.99%、90.98%、98.08%、115.58%；16批次热毒宁注射液中6个活性成分的平均质量浓度依次为2.00、26.63、52.63、5.29、34.64、9.69 μg/mL。结论 该方法简单准确，具有良好的重复性和稳定性，可为热毒宁注射液的质量标准的完善提供科学依据。

Objective An ultra high performance liquid chromatography coupled to triple quadrupole mass spectrometry (UPLC-QqQ-MS/MS) method has been established to simultaneously determine the contents of six bioactive constituents[japonicaside A (JA), L-phenylalanion secologanin B (PSB), luteolin, scopolamine, (1S,6R,7R,10R)-6-carboxy-10-methyl-α-methylene-1-(1-oxobutyl)-cyclohexane acrylic acid (CMCA), and 3α,5α-tetrahydrodeoxycordifoline lactam (TL)] from Reduning Injection. Methods This chromatographic separation was performed on an Agilent Zorbax SB-Aq C₁₈ (150 mm×2.1 mm, 3.5 μm) column with the mobile phase consisting of 0.1% formic acid water (A) and methanol (B) in a gradient elution (0.01-2.00 min, 5% B; 2.00-4.00 min, 5%-40% B; 4.00-11.00 min, 40%-95% B; 11.00-13.00 min, 95% B; 13.00-13.10 min, 95%-5% B; 13.10-14.00 min, 5% B) at a flow rate of 0.5 mL/min and the colunm temperature was set at 20℃. The analytes were detected using electrospray ionization (ESI) source by positive and negative ion monitoring mode. A triple quadrupole mass spectrometer was operated by ESI source in positive and negative ionization mode with multiple reaction monitoring for the detection of the six compounds. In the positive ion monitoring mode, the flow rate of Collision Gas (CAD) was 8 mL/min, the flow rate of Curtain Gas (CUR) was 20 mL/min, the temperature (TEM) was 500℃ and the Ion Spray Voltage (IS) was 4 500 V. In the negative ion monitoring mode, the flow rate of Collision Gas (CAD) was 8 mL/min, the flow rate of Curtain Gas (CUR) was 20 mL/min, the temperature (TEM) was 500℃ and the Ion Spray Voltage (IS) was -4 500 V. Results All calibration curves of the six components showed excellent linear regressions (r ≥ 0.999 0) within the test range. The average recoveries were 78.93%, 114.65%, 101.99%, 90.98%, 98.08%, and 115.58%, and the average contents of six bioactive constituents in 16 batches of Reduning Injection were 2.00, 26.63, 52.63, 5.29, 34.64, and 9.69 μg/mL, respectively. Conclusion The established method is rapid, accurate, and has high repeatability, which could provide scientific evidences for the quality control of Reduning Injection.

江苏省自然科学青年基金：基于活性成分追踪分离及快速检识分析新策略的中药复方药效物质基础研究（BK20140441）；江苏省博士后科研资助计划（1401171C）；“重大新药创制”科技重大专项（2013ZX09402203）；国家中药标准化项目（ZYBZH-C-JS-31）