ICP-MS法研究重度烧伤患者创面肉芽组织中微量元素的亚细胞分布

摘要

A method for simultaneous and quantitative determination of Cr,Mn,Fe,Co,Cu,Zn,Se and Cd elements in the subcellular fractions of nuclei,mitochondria,lysosome,microsome and cytosol of wound granulation tissue of severe burn pa-tients by octopole reaction system (ORS)inductively coupled plasma mass spectrometry (ICP-MS)was established.Using dif-ferential centrifugation,the sample is separated into different subcellular fractions.The subcellular fraction was digested by HNO3+H2 O2 with microwave digestion followed by dilution with ultrapure water then the above 8 trace elements in the solution were analyzed directly by ICP-MS.In the presented method,using ORS eliminates the polyatomic interferences caused by the matrixes.Rh as internal standard element was used to compensate matrix effect and signal drift.The detection limits of the 8 elements are in the range of 0. 72~33. 05 ng·L-1 ,and the RSD is less than 8. 4%.The results showed that the levels of some elements in subcellular fractions of wound granulation tissues were significantly different from those of normal skin tissues. ORS-ICP-MS is a useful tool for simultaneous determination of multi-elements in wound granulation tissue of severe burn pa-tients,and could be widely used in other biological samples analysis.%利用八极杆碰撞反应池（ORS）技术的电感耦合等离子体质谱（ICP-MS）法研究了Cr，Mn，Fe，Co， Cu，Zn，Se，Cd等元素在重度烧伤患者创面肉芽组织中的亚细胞分布。采用差速分级离心技术将样品分离获得细胞核、线粒体、溶酶体、微粒体和细胞上清液等亚细胞组分，使用混合酸 HNO3＋H2 O2并结合密闭微波消解对各亚细胞组分进行消解，分别选用ORS的氦碰撞模式和氢反应模式校正了质谱分析过程中的多原子离子干扰，以Rh为内标元素校正了基本效应和信号漂移现象。各待测元素的检出限在0．72～33．05 ng·L-1之间，相对标准偏差（RSD）小于8．4％。采用ORS-ICP-MS法可实现重度烧伤患者创面肉芽组织中微量元素亚细胞分布的准确分析，并可广泛应用于其他生物样品的分析，多数元素在创面肉芽组织和正常皮肤组织亚细胞组分的分布中存在显著性差异。