摘要
建立大鼠血液和脑组织液中游离型青藤碱、川芎嗪、加巴喷丁、扑热息痛、普瑞巴林和阿米替林的微透析液取样及HPLC-MS/MS检测方法。大鼠的颈静脉和脑纹状体植入微透析探针,分别以枸橼酸缓冲溶液和林格氏液为灌流液,以1.5μL·min^-1的灌流速度,以20 min的间隔收集血液和脑组织液微透析液样品,连续收集至给药后24 h。液相色谱分离采用C18反相色谱柱(HSS T32.5μm,2.1 mm×50 mm),流动相为甲醇/水(含0.05‰甲酸),流速为0.3 m L·min^-1梯度洗脱;质谱检测采用电喷雾离子源,正离子模式,多反应监测方式。定量离子对青藤碱、川芎嗪、加巴喷丁、扑热息痛、普瑞巴林、阿米替林和内标纳洛酮分别为330/181、137/80、172/154、152/110、160/142、278/233和328/310。方法经选择性、线性范围、基质效应、准确度、精密度、稳定性和探针回收率等考察确证适合大鼠血液和脑组织液微透析液中以上药物的测定。计算的微透析探针体内回收率在19.38%~25.88%之间。以40~50 mg·kg^-1的剂量大鼠股静脉注射给药,测定的血液微透析液中峰浓度在0.2~10μg·m L^-1内;脑组织液微透析液峰浓度在0.1~6μg·m L^-1内,但各时间点浓度水平均低于血液微透析液。总之,该方法成功用于大鼠给药青藤碱、川芎嗪、加巴喷丁、扑热息痛、普瑞巴林和阿米替林的微透析液取样和游离型药物浓度定量研究。
The treatment plan for chronic pain often proceeds from a single drug to drug combination therapy.Sinomenine and ligustrazine,natural alkaline substances derived from traditional Chinese medicines,are expected to provide a new choice for combination analgesic therapy strategies.Here we establish a microdialysis sampling and HPLC-MS/MS quantification method for sinomenine,ligustrazine,gabapentin,paracetamol,pregabalin and amitriptyline in rat blood and brain extracellular fluid.Blood and brain microdialysis probes were implanted in the jugular vein toward the right atrium and left corpus striatum zone(AP+0.2 mm,ML 3.0 mm,DV 3.5 mm)in rats.The blood and brain microdialysis probes were perfused with citric acid buffer solution and Ringer’s solution,respectively.Blood and brain extracellular fluid microdialysate were collected at intervals of 20 min at a perfusion rate of 1.5μL·min^-1,and continuously collected for 24 h after administration.The liquid chromatographic separation used a C18-reversed phase chromatographic column(HSS T32.5μm,2.1 mm×50 mm),the mobile phase was meth‐anol/water(containing 0.05‰formic acid),and gradient elution was carried out at a flow rate of 0.3 m L·min^-1.Mass spectrometric detection used an electrospray ion source,positive ion mode and multi-reaction monitoring method.The selected quantitative ions for sinomenine,ligustrazine,gabapentin,paracetamol,pregabalin,amitripty‐line and internal standard naloxone were 330/181,137/80,172/154,152/110,160/142,278/233 and 328/310 respec‐tively.The specificity,linear range,matrix effect,accuracy,precision,stability and probe recovery were investigated and confirmed to be suitable for the determination of the above drugs in rat blood and brain extracellular fluid microdialysate.The calculated in vivo recovery of microdialysis probes ranged from 19.38%to 25.88%.After intra‐venous administration of sinomenine(50 mg·kg^-1),ligustrazine(50 mg·kg^-1),gabapentin(50 mg·kg^-1),paracetamol(50 mg·kg^-1),pregabalin(50 mg·kg^-1)and amitripty
作者
李涛
赵小亮
高天乐
焦玥
高文雅
刘洋
张美玉
王志国
王丹巧
LI Tao;ZHAO Xiao-liang;GAO Tian-le;JIAO Yue;GAO Wen-ya;LIU Yang;ZHANG Mei-yu;WANG Zhi-guo;WANG Dan-qiao(Beijing Key Laboratory of Traditional Chinese Medicine Basic Research on Prevention and Treatment of Major Diseases,Experimental Research Center,China Academy of Chinese Medical Sciences,Beijing 100700,China;State Key Laboratory of Bioactive Substances and Function of Natural Medicine,Institute of Materia Medica,Chinese Academy of Medical Sciences,Beijing 100050,China;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)
出处
《药学学报》
CAS
CSCD
北大核心
2020年第9期2198-2206,共9页
Acta Pharmaceutica Sinica
基金
国家自然科学基金资助项目(81503278)
中央级公益性科研院所基本科研业务费专项资金资助项目(ZZ2019008,ZZ2018009)
中国中医科学院优秀青年基金资助项目(ZZ13-YQ-078)。
