Zarzycki Paweł K, Slączka Magdalena M, Włodarczyk Elżbieta, Baran Michał J
Section of Toxicology and Bioanalytics, Koszalin University of Technology, Śniadeckich 2, 75-453 Koszalin, Poland.
Chromatographia. 2013;76(19):1249-1259. doi: 10.1007/s10337-013-2445-3. Epub 2013 Mar 19.
In this work we demonstrated analytical capability of micro-planar (micro-TLC) technique comprising one and two-dimensional (2D) separation modes to generate fingerprints of environmental samples originated from sewage and ecosystems waters. We showed that elaborated separation and detection protocols are complementary to previously invented HPLC method based on temperature-dependent inclusion chromatography and UV-DAD detection. Presented 1D and 2D micro-TLC chromatograms of SPE (solid-phase extraction) extracts were optimized for fast and low-cost screening of water samples collected from lakes and rivers located in the area of Middle Pomerania in northern part of Poland. Moreover, we studied highly organic compounds loaded in the treated and untreated sewage waters obtained from municipal wastewater treatment plant "Jamno" near Koszalin City (Poland). Analyzed environmental samples contained number of substances characterized by polarity range from estetrol to progesterone as well as chlorophyll-related dyes previously isolated and pre-purified by simple SPE protocol involving C18 cartridges. Optimization of micro-TLC separation and quantification protocols of such samples were discussed from the practical point of view using simple separation efficiency criteria including total peaks number, log(product Δ), signal intensity and peak asymmetry. Outcomes of the presented analytical approach, especially using detection involving direct fluorescence (UV366/Vis) and phosphomolybdic acid (PMA) visualization are compared with UV-DAD HPLC-generated data reported previously. Chemometric investigation based on principal components analysis revealed that SPE extracts separated by micro-TLC and detected under fluorescence and PMA visualization modes can be used for robust sample fingerprinting even after long-term storage of the extracts (up to 4 years) at subambient temperature (-20 °C). Such approach allows characterization of wide range of sample components that are present in given extract in high and middle concentration range. Due to protocol simplicity and low cost of analysis this method can be useful for preliminary sample screening.
在本研究中，我们展示了微平面（微型薄层色谱）技术的分析能力，该技术包括一维和二维分离模式，可用于生成源自污水和生态系统水体的环境样品指纹图谱。我们表明，精心设计的分离和检测方案与先前发明的基于温度依赖性包合色谱和紫外二极管阵列检测的高效液相色谱方法相辅相成。所展示的固相萃取（SPE）提取物的一维和二维微型薄层色谱图经过优化，可用于快速、低成本地筛选从波兰北部中波美拉尼亚地区的湖泊和河流采集的水样。此外，我们研究了从波兰科沙林市附近的“亚姆诺”城市污水处理厂获得的经过处理和未经处理的污水中所含的高有机化合物。分析的环境样品包含多种物质，其极性范围从雌三醇到孕酮，以及先前通过涉及C18柱的简单固相萃取方案分离和预纯化的叶绿素相关染料。从实际角度出发，使用包括总峰数、log（乘积Δ）、信号强度和峰不对称性在内的简单分离效率标准，讨论了此类样品的微型薄层色谱分离和定量方案的优化。将所提出的分析方法的结果，特别是使用直接荧光（UV366/Vis）检测和磷钼酸（PMA）可视化的结果，与先前报道的紫外二极管阵列高效液相色谱生成的数据进行了比较。基于主成分分析的化学计量学研究表明，通过微型薄层色谱分离并在荧光和PMA可视化模式下检测的固相萃取提取物，即使在低于环境温度（-20°C）下长期储存提取物（长达4年）后，也可用于可靠的样品指纹识别。这种方法能够对给定提取物中高、中浓度范围内存在的多种样品成分进行表征。由于该方案简单且分析成本低，此方法可用于初步样品筛选。
