高效液相色普法测定兔灌胃后血清中百草枯浓度及其变化特征  

Paraquat concentration in rabbit serum determinated by HPLC after intragastric administration and its characteristics

作　　者：黄晓亮[1] 李映新[1] 凌家杰[2] 党毅 刘金远[3] 姚佩珊 颜池[2] 雷卓青 Xiao-liang Huang;Ying-xin Li;Jia-jie Ling;Yi Dang;Jin-yuan Liu;Pei-shan Yao;Chi Yan;Zhuo-qing Lei(Pharmaceutical College,Guangxi Medical University,Nanning,Guangxi 530021,China;School of Preclinical Medicine,Guangxi Medical University,Nanning,Guangxi 530021,China;Department of General Medicine,Guangxi Medical University,Nanning,Guangxi 530021,China;Emergency Department,the Second Affiliated Hospital of Guangxi Medical University,Nanning,Guangxi 530007,China)

机构地区：[1]广西医科大学药学院,广西南宁530021 [2]广西医科大学基础医学院,广西南宁530021 [3]广西医科大学全科学院,广西南宁530021 [4]广西医科大学第二附属医院西院急诊科,广西南宁530007

出　　处：《中国现代医学杂志》2018年第13期17-21,共5页China Journal of Modern Medicine

基　　金：广西医科大学救援人才小高地开放性课题(No:GXJZ201415)

摘　　要：目的探索建立高效液相色谱法(HPLC)在测定兔血清中百草枯浓度的可行性,并观察不同剂量百草枯灌胃后兔血清浓度的经时变化情况。方法 12只家兔按照随机数字表法分为两组,每组6只,分别灌胃给予100 mg/kg(高剂量组)和20 mg/kg(低剂量组)百草枯,于给药后1、2、5、12、24、48及72 h采集血清,血清用乙腈直接沉淀蛋白处理后进行高效液相测定。色谱柱条件:流动相为0.1 mol/L磷酸缓冲液(含12.5 mmol/L庚烷磺酸钠)-乙腈(90︰10,TFA调p H=2.8),流速为1.0 ml/min,检测波长为258 nm,柱温为30℃。结果所建立的方法在0.2~100.0 mg/L浓度范围内线性关系良好(r=0.999)。相对回收率在97.82%~101.20%之间,绝对回收率在84.37%~88.80%之间,日内和日间精密度(RSD)<5%。百草枯血药浓度时间曲线为单峰,无论是高剂量组还是低剂量组,给药2 h后其血清浓度达到峰值,之后逐渐下降,其中,高剂量组于给药后72 h后检测不出百草枯,低剂量组于药后24 h检测不出百草枯。重复测量设计的方差分析结果显示,不同时间点间、不同剂量组间的百草枯浓度及高、低剂量组随时间变化趋势差异有统计学意义。结论所建立的HPLC方法可用于测定兔血清百草枯浓度,不同剂量百草枯兔灌胃后其血清浓度和蓄积的时间与剂量有关,给药2 h后血药浓度达峰值。Objective To explore the feasibility of high performance liquid chromatography(HPLC)in determination of paraquat concentration in rabbit serum,and to observe the characteristics of the concentration changes after intragastric administration of paraquat of different dosages in rabbits.Methods Twelve New Zealand rabbits were randomly divided into two groups,with 6 rabbits in each group.The rabbits were orally given different dosages of paraquat to induce acute toxicosis,their serum was collected after administration for 1,2,5,12,24,48 and 72 h respectively and each serum sample was deproteinized with acetonitrile.The concentration of paraquat in serum was determinated by HPLC.The mobile phase was 0.1 mol/L phosphate buffer(12.5 mmol/L sodium heptanesulfonate)-acetonitrile(90:10),pH was adjusted to 2.8,the flow rate was 1.0 ml/min,the detector wavelength was 258 nm,column temperature was at 30℃.Results Excellent linear relationship was obtained within the range of 0.2-100.0 mg/L(r=0.999).The relative recovery was between 97.82%and 101.20%,and the absolute recovery was between 84.37%and 88.80%.The intra-day and inter-day RSD was over 5%.The plasma concentration timedependent reliability of Paraquat was unimodal curve with the peak value obtained 2 h after administration,then the concentration of paraquat in serum decreased.The concentration of paraquat could not be detected after 24 h in the low-dosage group and after 72 h in the high-dosage group.Variance of repeated measurement analysis revealed significant differences in the concentrations of paraquat among different time points(F=221.627,P=0.000),different dosage groups(F=133.618,P=0.000),and change trends with time of different dosage groups(F=81.874,P=0.000),respectively.Conclusions The established HPLC can be used to determine the concentration of paraquat in rabbit serum.Our results showed that the paraquat concentration and its accumulation time are positively correlated with the dosage,and the concentration reaches the peak value 2 h after administration.

关 键 词：百草枯 血清 高效液相色谱法 

分 类 号：R-332[医药卫生]