LC-MS/MS法测定Wistar大鼠组织中银杏内酯B的浓度  

Determination of ginkgolide B in rat tissues by LC-MS/MS

作　　者：刘冰洋[1] 刘有平[1] 王鑫[1] 王寅[1] 郝瑞家[1] 邸欣[1] 

机构地区：[1]沈阳药科大学药物代谢与药物动力学实验室,辽宁沈阳110016

出　　处：《沈阳药科大学学报》2012年第7期541-545,574,共6页Journal of Shenyang Pharmaceutical University

摘　　要：目的建立LC-MS/MS法测定大鼠组织中银杏内酯B的浓度,并将此方法应用于银杏内酯B注射液在Wistar大鼠体内的组织分布研究。方法色谱柱:AQVASIL C18柱(100 mm×2.0 mm I.D.,5μm),流动相:乙腈-水-甲酸(体积比为60.0∶40.0∶0.4),流速:0.4 mL.min-1,离子源为电喷雾电离(electrospray ionization,ESI)源,负离子化方式,以选择反应监测(selective reaction monito-ring,SRM)方式进行扫描定量,用于定量分析的离子反应分别为m/z 423→m/z 367(银杏内酯B)和m/z 444→m/z 138(格列吡嗪)。测定了18只Wistar大鼠静脉注射给予银杏内酯B(给药剂量为3.75 mg.kg-1)后大鼠体内各组织和脏器中银杏内酯B的质量浓度。结果测定大鼠组织匀浆液中银杏内酯B的线性为1.0～500.0μg.L-1,定量下限为1.00μg.L-1。低、中、高3个质量浓度质控样品的日内和日间精密度分别小于6.4%和14.5%,准确度为97.3%～103.6%。银杏内酯B在心、肝、脾、肺、肾、胰、胃、小肠、脑、肌肉、脂肪、子宫、卵巢和睾丸14种组织中(除脑组织6 h时间点外)均被检出。银杏内酯B在大鼠各组织中分布广泛,自各组织中消除也较快。结论该方法适用于银杏内酯B注射液在Wistar大鼠体内的组织分布研究。Objective To develop a convenient and sensitive method for the determination of ginkgolide B in Wistar rat tissues by LC-MS/MS for tissue distribution study.Methods Chromatographic separation was performed on an AQVASIL C18（100 mm×2.0 mm I.D.,5 μm）column with a mobile phase consisting of acetonitrile-water-formic acid（V∶ V∶ V=60.0∶ 40.0∶ 0.4）at the flow rate of 0.4 mL · min-1.A tandem mass spectrometer equipped with electrospray ionization（ESI）source was used as the detector and operated in the negative ion mode.Quantification was performed in selected reaction monitoring（SRM）mode using the precursor-production transitions of m/z 423→m/z 367 and m/z 444→m/z 318 for ginkgolide B and glipizide（I.S.）,respectively.The method was used to determine the concentration of ginkgolide B in Wistar rat tissues after given a single intravenous injection of EG-1 injector（3.75 mg · kg-1）.Results The linear range of ginkgolide B in Wistar rat tissue homogenate was 1.0-500.0 μg · L-1,and LLOQ was 1.00 μg · L-1.The intra-and inter-day precisions at three QC levels were less than 6.4% and 14.5%,respectively.The accuracy was within 97.3%-103.6%.Ginkgolide B was detected in all the fourteen tissues including heart,liver,spleen,lung,kidney,pancreas,stomach,intestine,brain,muscle,fat,uterus,ovary and testicle at 0.5 h,2 h and 6 h after administration（except for brain samples at 6 h）.Ginkgolide B distributed generally and eliminated quickly in rat tissues.No obvious accumulation was observed.The drug could permeate blood-brain barrier easily.Conclusions The method is simple,rapid and sensitive.It can be applied to Wistar rat tissue distribution studies of ginkgolide B.

关 键 词：银杏内酯B 液相色谱-串联质谱法 组织分布 

分 类 号：R972.9[医药卫生—药品] R917[医药卫生—药学]