Binder Grace A, Metcalf Rainer, Atlas Zachary, Daniel Kenyon G
University of South Florida, Tampa, Chemistry Department & Florida Center for Drug Discovery & Innovation (CDDI), USA.
University of South Florida, Tampa, Chemistry Department & Florida Center for Drug Discovery & Innovation (CDDI), USA.
Anal Biochem. 2018 Feb 15;543:37-42. doi: 10.1016/j.ab.2017.11.016. Epub 2017 Nov 24.
Digestion techniques for ICP analysis have been poorly studied for biological samples. This report describes an optimized method for analysis of trace metals that can be used across a variety of sample types. Digestion methods were tested and optimized with the analysis of trace metals in cancerous as compared to normal tissue as the end goal. Anthropological, forensic, oncological and environmental research groups can employ this method reasonably cheaply and safely whilst still being able to compare between laboratories. We examined combined HNO and HO digestion at 170 °C for human, porcine and bovine samples whether they are frozen, fresh or lyophilized powder. Little discrepancy is found between microwave digestion and PFA Teflon pressure vessels. The elements of interest (Cu, Zn, Fe and Ni) yielded consistently higher and more accurate values on standard reference material than samples heated to 75 °C or samples that utilized HNO alone. Use of HSO does not improve homogeneity of the sample and lowers precision during ICP analysis. High temperature digestions (>165 °C) using a combination of HNO and HO as outlined are proposed as a standard technique for all mammalian tissues, specifically, human tissues and yield greater than 300% higher values than samples digested at 75 °C regardless of the acid or acid combinations used. The proposed standardized technique is designed to accurately quantify potential discrepancies in metal loads between cancerous and healthy tissues and applies to numerous tissue studies requiring quick, effective and safe digestions.
用于电感耦合等离子体(ICP)分析的生物样品消化技术研究较少。本报告描述了一种优化的痕量金属分析方法,该方法可用于多种样品类型。以癌组织与正常组织中痕量金属分析为最终目标,对消化方法进行了测试和优化。人类学、法医学、肿瘤学和环境研究小组可以以合理的成本安全地采用这种方法,同时仍能够在不同实验室之间进行比较。我们研究了在170°C下对人、猪和牛的样品(无论其是冷冻、新鲜还是冻干粉末)进行硝酸(HNO₃)和过氧化氢(H₂O₂)联合消化的情况。发现微波消解和全氟烷氧基(PFA)特氟龙压力容器之间差异不大。与加热至75°C的样品或仅使用硝酸(HNO₃)的样品相比,感兴趣的元素(铜、锌、铁和镍)在标准参考物质上产生的数值始终更高且更准确。使用硫酸(H₂SO₄)并不能提高样品的均匀性,反而会降低电感耦合等离子体分析期间的精度。建议采用如所述的硝酸(HNO₃)和过氧化氢(H₂O₂)组合进行高温消解(>165°C),作为所有哺乳动物组织,特别是人类组织的标准技术,并且无论使用何种酸或酸组合,其产生的值比在75°C下消化的样品高300%以上。所提出的标准化技术旨在准确量化癌组织和健康组织之间金属负荷的潜在差异,并适用于众多需要快速、有效和安全消解的组织研究。