目的 建立高效液相色谱/质谱联用法(LC-MS/MS)同时测定健康受试者静脉滴注热毒宁注射液后血浆中金银花中成分绿原酸、新绿原酸、隐绿原酸, 栀子中成分栀子苷、山栀苷、京尼平龙胆双糖苷的质量浓度, 研究热毒宁注射液在健康人体内的药动学。方法 16名健康受试者静脉输注热毒宁注射液2支, 共20 mL(以5%葡萄糖注射液稀释至250 mL), 静脉输注时间定为90 min, 分别于输注前及输注过程中、输注后不同时间点取静脉血, 测定6种成分的血药浓度, 计算其药动学参数。测定绿原酸、新绿原酸、隐绿原酸的LC-MS/MS条件:Inertsil ODS-2色谱柱(150 mm×2.1 mm, 5 μm), 流动相为乙腈-0.1%甲酸, 卷帘气137.9 kPa(20 psi);碰撞气56.16 kPa(8 psi);喷雾电压-4 500 V。测定栀子苷、山栀苷和京尼平龙胆双糖苷的LC-MS/MS条件: Ecosil C₁₈色谱柱(150 mm×4.6 mm, 5 μm), 流动相为甲醇-20 mmol/L甲酸铵(含0.1%甲酸、10%甲醇), 卷帘气96.53 kPa(14 psi);碰撞气56.16 kPa(8 psi);喷雾电压4 500 V。结果 新绿原酸、绿原酸、隐绿原酸的t_max分别为1.31、1.38、1.44 h;C_max分别为1 241、3 294、2 121 ng/mL;AUC_0~t分别为1 972、5 351、3 596 ng·h/mL;MRT_0~t分别为0.708、0.790、0.899 h;CL分别为10.3、3.85、5.73 L/h;Vd分别为16.7、7.66、10.7 L;t_1/2分别为1.13、1.36、1.27 h。栀子苷、山栀苷、京尼平龙胆双糖苷的t_max分别为1.41、1.47、1.47 h;C_max分别为4.49、0.288、0.541 μg/mL;AUC_0~t分别为7.41、0.671、1.22 μg·h/mL;MRT_0~t分别为0.856、1.59、1.52 h;CL分别为2.74、30.0、16.6 L/h;Vd分别为5.63、60.9、35.2 L;t_1/2分别为1.42、1.42、1.47 h。结论 本试验建立的定量测定方法简便、准确, 可用于热毒宁注射液人体内药动学研究。

Objective To establish a high performance liquid chromatography/mass spectrometry (HPLC-MS/MS) for the simultaneous determination of the concentration in plasma of chlorogenic acid, new chlorogenic acid, cryptochlorogenic acid, gardenia component geniposide, shanzhiside, and genipin gentian diglucoside from Lonicerae Flos in Reduning Injection iv dripped in healthy human body. Methods Sixteen healthy subjects were iv infused of Reduning Injection with 2 ampoules, a total of 20 mL diluted with 5% glucose injection to 250 mL, iv injection time was set for 90 min, venous blood was measured at the different time points before, during, and after infusion, respectively. The conditions of determination for chlorogenic acid and other ingredients were as follows: Inertsil ODS-2 chromatographic column (150 mm × 2.1 mm, 5 μm), mobile phase (acetonitrile, 0.1% formic acid), rolling gas 137.9 kPa (20 psi), collision gas 56.16 kPa (8 psi), spray voltage -4 500 V. The LC-MS/MS conditions for determination of shanzhiside, geniposide and genipin gentian diglycoside were as follows: Ecosil C₁₈ (150 mm × 4.6 mm, 5 μm), the mobile phase (methanol, 20 mmol/L ammonium formate, 0.1% formic acid, and 10% methanol), rolling gas 96.53 kPa (14 psi), collision gas 56.16 kPa (8 psi), spray voltage 4 500 V. Results The t_max of neochlorogenic acid, chlorogenic acid, and cryptochlorogenic acid were 1.31, 1.38, and 144 h; C_max were 1 241, 329 4, and 2 121 ng/mL; AUC_0~t were 1 972, 5 351, and 3 596 ng·h/mL; MRT_0~t were 0.708, 0.790, and 0.899 h; CL were 10.3, 385, and 5.73 L/h; Vd were 16.7, 766, and 10.7 L; t_1/2 were 1.13, 1.36, and 1.27 h, respectively. The t_max of geniposide, shanzhiside, and genipin gentian diglucoside were 1.41, 1.47, and 1.47 h; C_max were 4.49, 0.288, and 0.541 μg/mL; AUC_0~t were 7.41, 0.671, and 1.22 μg·h/mL; MRT_0~t were 0.856, 1.59, and 152 h; CL were 2.74, 30.0, and 16.6 L/h; Vd were 5.63, 60.9, and 35.2 L; t_1/2 were 1.42, 1.42, and 1.47 h. Conclusion The method is simple, accurate, and can be used for in vivo pharmacokinetic study on Reduning Injection.

"十二五"重大新药创制项目"儿科中药新药临床评价研究技术平台规范化建设"课题(2011ZX09302-006-03)