【摘要】 目的:研究大鼠尿样中多种马兜铃酸类(AAs)及马兜铃内酰胺类(ALs)化合物的固相萃取和富集的方法,为定量测定建立前处理方法。方法:以马兜铃酸-Ⅰ(AA-Ⅰ)、马兜铃酸-Ⅱ(AA-Ⅱ)、马兜铃内酰胺-Ⅰ(AL-Ⅰ)、马兜铃内酰胺-Ⅱ(AL-Ⅱ)和其他16个AAs及ALs化合物的高效液相色谱峰面积为指标,比较填料为C18的Agilent C18/100 mg,Alltech HC18/100 mg,Alltech C18/100 mg,Alltech C18/300 mg和填料为苯基(Phenyl)的AgilentPhenyl/200 mg 5种固相萃取(SPE)小柱对目标成分的萃取保留作用的强弱;以AA-Ⅰ,AA-Ⅱ,AL-Ⅰ和AL-Ⅱ的萃取回收率为指标,比较水和1%醋酸-0.02%三乙胺水溶液2种淋洗溶剂,乙醚、丙酮、氯仿、醋酸乙酯、二氯甲烷、甲醇、乙腈7种洗脱溶剂对AAs及ALs化合物的萃取效果的影响;采用4因素3水平正交试验对影响固相萃取的条件包括活化体积、清洗体积、淋洗体积、洗脱体积等进行优化,以AA-Ⅰ,AA-Ⅱ,AL-Ⅰ,AL-Ⅱ和其他7个分离度好的AAs及ALs的色谱峰面积为指标,确定最佳的因素水平。结果:确定固相萃取的条件为采用200 mg的Phenyl柱作固相萃取小柱,用1.0 mL甲醇活化和1.0 mL水清洗后,上大鼠尿样1.0 mL,以1%醋酸-0.02%三乙胺水溶液0.8 mL淋洗,3.0 mL甲醇洗脱。结论:建立的尿样固相萃取方法萃取效率高、选择性好、简单、省时,可作为大鼠尿样中多种AAs及ALs化合物分析测定的尿样前处理方法。更多还原

【Abstract】 Objective: To develop a urine pretreatment method of Solid Phase Extraction(SPE) for the quantitative determination of a number of aristolochic acids(AAs) and aristololactams(ALs) in rat urine.Method: The HPLC peak area of AA-Ⅰ,AA-Ⅱ,AL-Ⅰ and AL-Ⅱ,and other sixteen AAs and ALs was chosen as evaluating index to study the extract results of five Solid Phase Extraction columns(Agilent C18/100 mg,Alltech HC18/100 mg,Alltech C18/100 mg,Alltech C18/300 mg and Agilent Phenyl/200 mg) comparatively.The influences of two washing solvents(water and 1% acetic acid-0.02% triethylamine solution) and seven eluting solvents(ether,acetone,chloroform,ethyl acetate,dichloromethane,methanol and acetonitrile) on extract results of AAs and ALs are comparatively studied with the extracting recoveries of AA-Ⅰ,AA-Ⅱ,AL-Ⅰ and AL-Ⅱ as indicators.The HPLC peak area of AA-Ⅰ,AA-Ⅱ,AL-Ⅰ and AL-Ⅱ,and other seven AAs and ALs with good separation being targets,several factors which affect extracting efficiency of analytes,including activating volume,cleansing volume,washing volume and eluting volume,are optimized by orthogonal design experiments with four factors at three levels.Result: The established method of SPE is as follows: Agilent Phenyl SPE column of 200 mg,activating with 1.0 mL methanol,cleansing with 1 mL water,adding 1.0 mL rat urine sample,washing with 0.8 mL 1% acetic acid 0.02% triethylamine solution,and eluting with 3.0 mL methanol.Conclusion: The established method of SPE is efficient,selective,simple and fast,and can be used as urine pretreatment method to analyze a variety of aristolochic acids and aristololactams in rat urine.更多还原