摘要
目的:建立应用荧光偏振免疫分析法(FPIA)测定小鼠血药浓度的实验方法,探讨该方法测定庆大霉素、地高辛和丙戊酸钠3种常见小分子药物小鼠血药浓度的可行性。方法:选用庆大霉素、地高辛和丙戊酸钠标准品,采用荧光偏振(FP)值非线性拟合法确定药物检出限及线性范围,在线性范围内分别设置低、中和高3个浓度组,检测质控偏差、日间精密度和小鼠血清干扰水平。选用30只KM小鼠进行药物代谢曲线测试,分为庆大霉素组(肌肉注射5μg·g-1)、地高辛组(灌胃20ng·g-1)和丙戊酸钠组(灌胃15μg·g-1),每组10只,给药后24h内取5个时间点(1、3、6、12和24h)测定小鼠血药浓度。结果:FPIA法检测3种药物的检出限均在50μg·L-1以下,线性范围较宽(大于10倍检出限),检测质控误差小于5%,日间精密度较高[中和高浓度组变异系数(CV)<5%],小鼠血清对庆大霉素和地高辛的检测效果无明显影响[误差百分比(ER)<5%],高浓度血清对丙戊酸钠的检测效果有一定的负向影响(50%血清组ER为-12.84%)。小鼠给药1h后庆大霉素达到血药浓度峰值,半消期3h;1h后地高辛达到血药浓度峰值,半消期>24h;3h后丙戊酸钠达到血药浓度峰值,半消期6~12h。结论:FPIA法测定小分子药物小鼠血药浓度所需样本量小,精确度高,前处理简单,具有高通量和快速检测的优势特点,可广泛应用于药理学、药剂学和临床检测研究中。
Objective：To establish a drug concentration in plasma determination method based on fluorescence polarization immunoassay（FPIA）,and to explore the feasibility of this method in monitoring the concentrations of three small molecular drugs including gentamycin,digoxin and sodium valproate in plasma of the mice.Methods：The standard samples of gentamycin,digoxin,and sodium valproate were selected.The limits and linear ranges of drugs were measured with fluorescence polarization（FP）non-linear fitting method.Three standard groups with low,medium,and high concentrations of drug were selected during liver range,and the accuracy,inter-day precisions and serum anti-interference abilities of the mice were detected.Total 30 KM mice were divided into gentamycin group（5μg·g-1,intramuscular injection）,digoxin group（20 ng·g-1,gavage）and sodium valproate group（15μg·g-1,gavage）;the blood drug concentrations of the mice were measured at 1,3,6,12 and 24 htime points after administration.Results：For all three drugs detected,the detection limits detected by FPIA were lower than 50μg·L-1）,the linear ranges were broad（more than 10 times of detection limit）,the accuracy（ER）was lower than 5% and the inter-day precisions were higher（CV5% in medium and high concentration groups）.The mouse serum showed no affection to FPIA detection qualities for gentamycin and digoxin,while high concentration of serum negatively affected the detection quality for sodium valproate（ER=-12.84%in 50% serum group）.After administration,gentamycin reached the peak blood concentration at 1 h with half-life period of 3 h,digoxin reached the peak blood concentration at 1 hwith half-life period of 〉24 h,and sodium valproate reached the peak blood concentration at 3 hwith half-life period of 6-12 h.Conclusion：FPIA for small molecular drug concentration in plasma of the mice has the advantages of small sample size,high accuracy,simple preprocessing,high throughput detection and rapid test, which can be applied in de
作者
刘洋
刘艳
刘宸辛
冯志桐
姜丹
LIU Yang;LILT Yah;LIU Chenxin;FENG Zhitong;JIANG Dan(School of Life Science,Jilin University,Changchun 130021,China)
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2018年第4期833-838,共6页
Journal of Jilin University：Medicine Edition
基金
吉林省科技厅科技创新人才培育计划资助课题(20140520005JH)
吉林大学实验技术项目资助课题(409020720134)
关键词
荧光偏振免疫分析
血药浓度
庆大霉素
地高辛
丙戊酸钠
fluorescence polarization immunoassay
drug plasma concentration
gentamycin
digoxin
sodium valproate
