液液萃取-GFAAS法测定生物样品中的Cu(Ⅰ)  

Determination of Cuprous in Biological Samples by Liquid-Liquid Extraction-GFAAS

作　　者：张源 吴鹏 李慧 罗红军 罗文鸿 林哲绚 ZHANG Yuan;WU Peng;LI Hui;LUO Hong-jun;LUO Wen-hong;LIN Zhe-xuan(Laboratory of Biological Analysis of Shantou University Medical College,Shantou 515041,China)

机构地区：[1]汕头大学医学院生物分析实验室

出　　处：《光谱学与光谱分析》2020年第2期632-636,共5页Spectroscopy and Spectral Analysis

基　　金：国家自然科学基金项目(31671183);广东省自然科学基金项目(2018A030307005)资助

摘　　要：建立液液萃取-石墨炉原子吸收法测定生物样品中Cu(Ⅰ)方法。200μL血清及细胞匀浆、细胞膜液等样本与200μL 30%三氯乙酸混匀后,离心去蛋白,取上清液400与1500μLpH为12.5的甘氨酸-NaOH-缓冲溶液混至pH约为9,加入含1000μL 0.05%2,2′-联喹啉的正戊醇溶液,旋涡振荡1 min,静置分层后取有机层500μL置于2 mL特氟隆消化管,于95℃烘箱烘6 h挥发有机溶剂,冷却至室温后分别加入200μL硝酸和双氧水,80℃水浴消化,室温加入600μL 1%硝酸后用石墨炉原子吸收法,测定的结果为Cu(Ⅰ)含量。另取100μL血清(200μL细胞匀浆、细胞膜液)置于2 mL特氟隆消化管,于80℃烘箱烘5 h至样品干燥,冷却至室温分别加入200μL硝酸和双氧水,80℃水浴消化后,室温用1%硝酸稀释成1000μL,用石墨炉原子吸收法测定总铜含量。方法检测限:0.04μg·L-1,相对偏差<5%。回收率:95%~102%。用此法检测了一些无机样品和生物样品。测定的结果表明,宫颈癌患者血清Cu(Ⅰ)比正常人要高,总铜含量差别不大。一些无机样品如自来水、农夫山泉水、尿液含Cu(Ⅱ)离子不含Cu(Ⅰ)离子,而生物样品如肝细胞及肝细胞膜含有Cu(Ⅰ)离子不含Cu(Ⅱ)离子。本法可以在混有Cu(Ⅱ)离子的情况下测定Cu(Ⅰ)离子,且10倍Cu(Ⅱ)离子浓度存在下测定痕量Cu(Ⅰ)离子无干扰。A method for the determination of cuprous[Cu(Ⅰ)]in biological samples by liquid-liquid extraction-inductively coupled plasma mass spectrometry(ICP-MS)was developed.Serum and cell homogenate,cell membrane components were deproteinized with trichloroacetic acid,then the supernatant was mixed with a glycine-sodium hydroxide(NaOH)buffer(pH 12.5)to obtain a mixture with pH of 9.Then 1000μL of 0.05%2,2’-biquinoline dissolved in N-pentanol was added and vortexed for 1 min.After static layering,500μL of the organic layer was collected in a 2 mL Teflon digestive tube,and the organic solvent was evaporated in an oven at 95℃.After digestion with nitric acid and hydrogen peroxide,the sample solution was subjected to inductively coupled plasma mass spectrometry and the data of cuprous content were obtained.The detection limit of this method was 0.04μg·L-1,with relative deviation being less than 5%.The recovery was 95%to 102%.Then the method was applied to determine Cu(Ⅰ)of serum from cervical cancer patients and several types of water.The results showed that serum Cu(Ⅰ)was higher in cervical cancer patients than that in normal controls.Cuprous ions could not be detected in tap water,Nongfu Spring water,and urine.While,Cuprous ions but not divalent copper ions could be detected in cytosol and cell membrane.This method could detect trace Cu(Ⅰ)ions in the presence of 10 times concentration of Cu(Ⅱ)ion without interference.

关 键 词：亚铜 石墨炉原子吸收法光谱 甘氨酸 2 2′-联喹啉 

分 类 号：O657.3[理学—分析化学]