生物样品中新污染物筛查:透析和凝胶渗透色谱联用净化方法  

Screening of New Pollutants in Biological Samples:A Combined Purification Method Based on Dialysis and Gel Permeation Chromatography

作　　者：娄小涵 佟宇俊 程飞[1] 游静[1] LOU Xiao-han;TONG Yu-jun;CHENG Fei;YOU Jing(College of Environment and Climate,Jinan University,Guangzhou 511443,China)

机构地区：[1]暨南大学环境与气候学院,广东广州511443

出　　处：《分析测试学报》2024年第8期1212-1219,共8页Journal of Instrumental Analysis

基　　金：广东省基础与应用基础研究基金项目(2023A1515011396)。

摘　　要：以河蚬为代表,建立了一种能有效去除生物样品中脂肪及其它内源性干扰物,并保留性质广泛的新污染物的净化方法。选择理化性质迥异的35种污染物(log Kow为-1.30~8.41),优化生物样品萃取、透析及凝胶渗透色谱(GPC)分离方法,并验证了方法可行性。优化的样品前处理方法为:样品先后采用正己烷、丙酮、二氯甲烷混合溶液(体积比2∶2∶1)和乙腈加速溶剂萃取,提取液经4次透析后再用GPC净化,目标物使用气相色谱-质谱和液相色谱-串联质谱进行分析。采用香兰素-磷酸法测定每一净化步骤后样品的脂肪含量,并结合色谱-质谱全扫描分析,检验干扰物的去除效率。结果表明,净化后河蚬提取物中残余的脂肪含量小于1.06%,GPC中待测分析物的流出时间为8~20 min,平均回收率为55.1%±13.0%,相对标准偏差为1.4%~9.2%。净化后的河蚬生物样品中脂肪和部分内源性物质被有效去除,宽泛程的目标物的回收率均达到环境样品痕量新污染物检测的要求,所建方法可用于生物样品非目标筛查的前处理。In the present study,a cleanup method using a combination of dialysis and gel perme⁃ation chromatography(GPC)was developed to remove lipids as well as natural hormones from biologi⁃cal samples with the Asian clam(Corbicula fluminea)as the model organism.To effectively screen new pollutants across different categories in biological samples,the procedures of sample extraction,dialysis,and GPC separation were optimized.This method effectively removed lipids and other en⁃dogenous interferences from biological samples while retaining a broad range of new pollutants.We selected 35 contaminants with diverse physicochemical properties(logKow ranging from-1.30 to 8.41)and evaluated their recoveries after each processing step and measured the lipid contents throughout the entire cleanup procedure.The optimized extraction method was to use accelerated sol⁃vent extraction(ASE)with a mixture of hexane,acetone and dichloromethane(2∶2∶1,volume ra⁃tio)and acetonitrile,sequentially.The extracts were subjected to four rounds of dialysis,and then were purified using GPC.Depending on their physicochemical properties,target analytes were ana⁃lyzed by gas chromatography-mass spectrometry(GC-MS)or liquid chromatography-tandem mass spectrometry(LC-MS/MS).Furthermore,a vanillin-phosphoric acid method was applied to measure lipid content after each purification step.At the same time,removal efficiency of the interferences was also verified using chromatography-mass spectrometry analyses in scan mode.Results after the combined dialysis and GPC treatment showed the residual lipid content in sample extraction was less than 1.06%.The elution time for target analytes in GPC was determined to be between 8 to 20 min.Average recoveries was 55.1%±13.0%,with relative standard deviations of 1.4%–9.2%.In conclusion,the developed method effectively removed lipids and some endogenous substances from clam ex⁃tracts.Meanwhile,the recoveries of all target analytes have met the requirements for trace environ⁃mental sample analysis,sh

关 键 词：生物样品 透析 凝胶渗透色谱 净化 非目标分析 

分 类 号：O657.7[理学—分析化学] X132[理学—化学]