液相色谱-串联质谱法同时测定大鼠血浆中咪达唑仑和非那西丁及其药动学研究

发布时间：2019-04-23 14:01

【摘要】：目的建立液相色谱-串联质谱法(LC-MS/MS)同时测定非那西丁(PN)、咪达唑仑(MDZ)在大鼠血液中的含量。方法大鼠随机被分为PN组、MDZ组和PN-MDZ合用组(均n=6),分别尾静脉注射PN、MDZ及PN和MDZ混合探针药物,剂量均为1 mg·kg-1。于指定时间眼眶采集血样,以苯海拉明为内标,采用LC-MS/MS测定大鼠血浆中探针药物及其代谢产物的浓度。色谱柱为Kinetex XB-C18柱(100 mm×3.0 mm,2.6μm),流动相为甲醇∶0.025%甲酸水,进行梯度洗脱。电喷雾离子源,以多反应离子监测方法进行正离子扫描,PN和其代谢产物醋氨酚(Ace),MDZ和其代谢产物1-羟基咪达唑仑(1-OH-MDZ)及苯海拉明离子对分别为m/z 180.2→110.0,m/z 152.2→110.1,m/z 326.2→291.2,m/z 342.2→324.2和m/z 256.3→167.2。结果PN、Ace、MDZ和1-OH-MDZ线性范围分别为:4.288~21 440 ng·m L-1、1.038~5 190 ng·m L-1、4.664~11 660 ng·m L-1、0.01~50 ng·m L-1;回收率、稳定性和日内、日间精密度均符合生物样品分析要求;PN和MDZ单用与合用前后药动学参数无显著差异(P0.05)。PN单用和与MDZ合用后t1/2分别为(0.44±0.15)、(0.42±0.08)h,ρmax分别为(9.35±1.58)、(10.17±0.76)μg·m L-1,AUC0-6 h分别为(4.21±0.63)、(4.90±0.42)μg·h·m L-1;MDZ单用和与PN合用后t1/2分别为(0.64±0.09)、(0.68±0.05)h,ρmax分别为(3.48±0.51)、(3.01±0.64)μg·m L-1,AUC0-6 h分别为(2.58±0.41)、(2.08±0.29)μg·h·m L-1。结论建立的测定方法可用于PN、MDZ以及其代谢产物的药动学研究,并证明PN、MDZ在大鼠体内基本没有代谢的相互作用。
[Abstract]:Objective to establish a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of fenacetin (PN), midazolam (MDZ) in rat blood. Methods the rats were randomly divided into three groups: PN group, MDZ group and PN-MDZ combination group (all n = 6). PN,MDZ and PN and MDZ mixed probe drugs were injected intravenously into tail vein at a dose of 1 mg kg-1.. Blood samples were collected from the orbit at the specified time and diphenhydramine was used as the internal standard to determine the concentration of probe drugs and their metabolites in rat plasma by LC-MS/MS. The chromatographic column was Kinetex XB-C18 column (100 mm 脳 3.0 mm, 2.6 渭 m), the mobile phase was methanol: 0.025% formic acid water, and gradient elution was carried out. Electrospray ion source, positive ion scanning with multi-reaction ion monitoring method, PN and its metabolite acetaminophen (Ace), MDZ and its metabolites, 1-hydroxymidazolam (1-OH-MDZ) and diphenhydramine ion pairs were 180.2, 152.2, 326.2 and 291.2, respectively. M\ x {5ee5} z 342.2 / 324.2 and m\ {# * $} z 256.3 / 167.2. Results the linear ranges of PN,Ace,MDZ and 1-OH-MDZ were 4.288 ~ 21,440 ng 路mL ~ (- 1), 1.038 ~ 5 190 ng 路mL ~ (- 1), 4.664 ~ 11.660 ng 路mL ~ (- 1), 0.01 ~ 50 ng 路mL ~ (- 1), respectively. The recovery, stability, intra-day and inter-day precision all meet the requirements of biological sample analysis. There was no significant difference in pharmacokinetic parameters before and after combination of PN and MDZ (P0.05). After PN alone and combined with MDZ, t _ 1-max _ 2 was (0.44 卤0.15), (0.42 卤0.08 h and 蟻 max was (9.35 卤1.58) h, respectively. (10.17 卤0.76) 渭 g 路mL ~ (- 1) and (4.21 卤0.63), (4.90 卤0.42) 渭 g 路h 路mL ~ (- 1) for 6 h, and (4.21 卤0.63) 渭 g 路h 路mL ~ (- 1) for 6 h, respectively. T _ 1 ~ (2), 蟻 _ (max) and AUC _ (0) were (0.64 卤0.09), (0.68 卤0.05h, (3.48 卤0.51), (3.01 卤0.64) 渭 g 路mL ~ (- 1) and (2.58 卤0.41) 渭 g 路mL ~ (- 1) for MDZ alone and combined with PN, respectively. (2.08 卤0.29) 渭 g 路h 路mL. Conclusion the established method can be used to study the pharmacokinetics of PN,MDZ and its metabolites, and it is proved that there is no metabolic interaction between PN,MDZ and its metabolites in rats.
【作者单位】：江西中医学院现代中药制剂教育部重点实验室;成都中医药大学药学院;
【基金】：国家自然科学基金资助项目(81060347) 国家留学基金委资助项目(留金法[2012]5031) 江西省青年科学学培养计划项目(20122BCB23021) 江西省教育厅科学技术研究项目(GJJ13606)
【分类号】：R965

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