Analysis of Selected Endocrine Disrupters Fraction Including Bisphenols Extracted from Daily Products, Food Packaging and Treated Wastewater Using Optimized Solid-Phase Extraction and Temperature-Dependent Inclusion Chromatography.

The aim of this research is to demonstrate the concept and ability for the fast and preliminary screening of complex food and environmental samples for the presence of endocrine disrupters fractions, consisting of low-molecular mass micropollutants, particularly various bisphenols (A, B, C, E, F, S, Z, AF, AP, BP and FL). The developed analytical protocol for this research requires two main steps: (i) optimized solid phase extraction (SPE) for selective isolation, purification and pre-concentration of target fraction, and (ii) selective temperature-dependent inclusion chromatography for samples analysis via a HPLC-UV-VisDAD system using isocratic elution and internal standard quantification approach. The chromatographic experiment revealed that both ß-CD and its hydroxypropyl derivative strongly interact with selected bisphenols. This is in contrast to the steroids and PAHs molecules investigated previously, where a strong interaction with ß-cyclodextrin was observed. Integrated areas derived from acquired chromatographic profiles for each individual sample were used as the simple classification variable enabling samples comparison. We demonstrated that the proposed analytical protocol allows for fast estimation of EDC fractions in various daily use products, food and environmental samples. The materials of interest were selected due to the presence in surface water ecosystems of their residues, and finally, in raw wastewater including rice bags, plastic bags, cloths, sanitary towels, fish baits and various plastic foils from food products. Treated sewage water released directly to the environment from a municipal treatment plant (Jamno, Koszalin) was also investigated. It has been demonstrated that a whole range of low-molecular mass compounds, which may be detected using UV-Vis detector, can easily be emitted from various in daily use products. The presence of micropollutants in treated wastewater, water ecosystems and plastic waste utilization via technological wastewater treatment processes must be addressed, especially in terms of microplastic-based pollutants acting as endocrine disrupters. It is hoped that the proposed simple analytical protocol will be useful for fast sample classification or selection prior to advanced targeted analysis involving the more accurate quantification of specific analytes using e.g., mass spectrometry detectors.

Unexpected differences between planar and column liquid chromatographic retention of 1-acenaphthenol enantiomers controlled by supramolecular interactions involving ß-cyclodextrin at subambient temperatures.

We report the results of experimental work focusing on host-guest supramolecular complex creation between macrocyclic compound (ß-cyclodextrin) and 1-acenaphthenol enantiomers (racemic mixture) in liquid phase composed of 35% acetonitrile in water (v/v) at different temperatures ranging from 0 to 90 °C. Experimental setup involved several analytical protocols based on classical non-forced flow planar chromatography (RP-18 TLC plates), micro-TLC (RP-18 W HPTLC plates), column chromatography (HPLC with C-18 and C-30 stationary phases), as well as UV-Vis spectrophotometry and optical microscopy. It has been found that under various planar chromatographic conditions (stationary plates type, chamber shape and volume, development mode, and saturation) non-typical retention properties (extremely high retention) of 1-acenaphthenol at subambient temperatures can be observed. To our knowledge, reported experimental results are in opposition to currently described retention models based on column chromatographic investigation of host-guest complexes (where in case of strong interaction of given analyte with macrocyclic mobile phases additive, which itself is non strongly retarded by stationary phase-close to the retention of dead volume marker, the retention of target compounds is shortened at low temperatures). To explain this TLC phenomenon that may have in our opinion a number of practical applications, especially for selective high throughput separation involving microchromatographic and/or microfluidic devices as well fractionation and extraction protocols (using, e.g., bar extraction systems), several experiments were conducted focusing on (i) acenaphthenol chromatography under different instrumental conditions, (ii) cyclodextrin retention measured as analyte or mobile phase additive, (iii) plate development time under different mobile phases and temperature settings, (iv) various column chromatographic conditions including C-30 and two C-18 stationary phases, (v) UV-Vis spectrophotometry, and (vi) microscopy inspection of precipitated CD-acenaphthenol crystals. Analysis of collected data has revealed that the most probable reasons for TLC retention behavior of 1-acenaphthenol under ß-cyclodextrin additive conditions can be associated with (i) solubility changes of created host-guest complex, (ii) kinetics of solid complex precipitation, and (iii) differences in analysis time between planar and column chromatography. Because precipitation phenomenon may have a massive impact on analytes quantification involving macrocycles as the mobile phase additives, our previously reported data concerning a number of low-molecular compounds (mainly steroids and non steroidal endocrine disrupting chemicals) using HPLC methodology based on binary mobile phases without and with ß-cyclodextrin and its hydroxypropyl derivative were re-examined and results discussed. Considering these data and the whole data set reported presently, the enhanced model of chromatographic retention driven by host-guest interaction was proposed.

Supplementary evaluation of retention and physicochemical data involving multivariate analysis approach.

This commentary highlights the issue of real differences between stationary phases that were studied in an experimental paper entitled "Novel stationary phases based on asphaltenes for gas chromatography" prepared by Grzegorz Boczkaj and co-authors (J. Sep. Sci. 2016, 39, 2527-2536). Particularly, a chemometric study has revealed relatively small differences between stationary phases investigated. Moreover, simple principle component analysis calculations enabled the identification of the outlier points within large raw dataset and to find the parameters (variables) that may carry equal information.

Estimation of the breakthrough volume of selected steroids for C-18 solid-phase extraction sorbent using retention data from micro-thin layer chromatography.

SPE method is a very popular technique, and is commonly used for the prepurification, concentration, and isolation of different organic compounds from variable matrices. In this work, the optimization of SPE process was carried out. The breakthrough volume of solid sorbents based on octadecylsilane was determined and three methods were compared: (1) calculation one - the breakthrough volume was calculated using retention factor k determined with micro-TLC method, frontal analysis - (2) breakthrough volume was determined as volume of whole elution peak, and (3) breakthrough volume was determined as the center of peak gravity. For calculation method, the k values of key estrogens and progestogens were derived from the micro-TLC experiment reported previously. By combining these three methods, we can point the start of elution, the maximum concentration of analyte in eluate, and the whole eluent volume, which is necessary to achieve an appropriate selectivity and high extraction recovery. Proposed calculation method allows to estimate the beginning of the steroid peak, when the analyte appears in the eluate flowing from the sorbent. Such observation advances the SPE optimization protocol that was described before and was based on the correlation between raw k(SPE) and k(micro-TLC) data.

Microfluidic Paper-Based Analytical Devices (µPADs) and Micro Total Analysis Systems (µTAS): Development, Applications and Future Trends.

Microfluidic paper-based analytical devices and micro total analysis systems are relatively new group of analytical tools, capable of analyzing complex biochemical samples containing macromolecules, proteins, nucleic acids, toxins, cells or pathogens. Within one analytical run, fluidic manipulations like transportation, sorting, mixing or separation are available. Recently, microfluidic devices are a subject of extensive research, mostly for fast and non-expensive biochemical analysis but also for screening of medical samples and forensic diagnostics. They are used for neurotransmitter detection, cancer diagnosis and treatment, cell and tissue culture growth and amplification, drug discovery and determination, detection and identification of microorganisms. This review summarizes development history, basic fabrication methods, applications and also future development trends for production of such devices.

Micro-TLC Approach for Fast Screening of Environmental Samples Derived from Surface and Sewage Waters.

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 ΔhRF), 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.

Long-Term Fluorescence Behavior of CdSe/ZnS Quantum Dots on Various Planar Chromatographic Stationary Phases.

Nanoparticles, particularly quantum dots (QDs), are commonly used for the sensitive detection of various objects. A number of target molecules may be determined using QDs sensing systems. Depending on their chemical nature, physicochemical properties, and spatial arrangement, QDs can selectively interact with given molecules of interest. This can be performed in complex systems, including microorganisms or tissues. Efficient fluorescence enables low exposure of QDs and high sensitivity for detection. One disadvantage of quantum dots fluorophores is fluorescence decay. However, for given applications, this property may be an advantage, e.g., for highly sensitive detection based on correlation images in the time domain. This experimental work deals with the measurement of fluorescence decay of Lumidot TMCdSe/ZnS (530 nm) quantum dots. These nanoparticles were transferred to the surface of various planar chromatographic stationary phases. Fluorescence of formed spots was recorded at room temperature over a long period of time, namely 15.7824 × 105 min (three years). The resulting signal profiles in the time domain were analyzed using classical approach (luminescence model comparison involving different mathematical models).Moreover, fluorescence behavior on different TLC/HPTLC supports was investigated using multivariate statistics (principal component analysis, PCA). Eight planar chromatographic stationary phases were investigated, including cellulose, octadecylsilane, polyamide, silica gel and aluminium oxide in different forms (TLC and HPTLC types). The presented research revealed significantly different and non-linear long-term QDs behavior on these solids. Two different fluorescence signal trajectories were recorded, including typical signal decay after QDs application to the plates and long-term intensity increase. This was particularly visible for given planar chromatographic adsorbents, e.g., cellulose or octadecylsilane. To the author's knowledge, these findings were not reported before using the stationary chromatographic phases, and enable the design of future experiments toward sensing of low molecular mass chemicals using, e.g., advanced quantification approaches. This may include signal processing computations based on correlation images in the time domain. Additionally, the reported preliminary data indicates that the investigated nanoparticles can be applied as efficient and selective fluorophores. This was demonstrated on micro-TLC plates where separated bioactive organic substances quenching from cyanobacteria extracts were sensitively detected. The described detection protocol can be directly applied for different planar chromatographic systems, including paper-based microfluidic devices, planar electrophoresis and/or miniaturized microfluidic chip devices.

Investigation of Hybrid Methods for Elimination of Brilliant Blue Dye from Water Phase Using Various Nanomaterials Combined with Activated Sludge and Duckweed.

The main goal of this experimental work is screening of different natural and synthetic nanomaterials and biopolymers that may improve elimination of stable micropollutants from water phase. In this work, as a target chemical acting as the micropollutant molecule, the Brilliant Blue (BB) dye was selected. We tested different active matrices dispersed in water phase including activated carbon (AC), lyophilized graphene oxide (GO), ß-cyclodextrin (CD), raw dandelion pappus (DP), microcrystalline cellulose(MC), and raw pine pollen (PP), as well as two types of Egyptian Blue mineral pigments (EB1 and EB2). Graphene oxide and Egyptian Blue nanomaterials were synthesized in our laboratory. We investigated potential application of such nanoparticles and biopolymer conglomerates as additives that may tune the activated sludge (AS) microorganisms or duckweed water plant (DW) and increase efficiency of micropollutants removal from wastewater. Studied nanomaterials/biopolymers were used in two different experimental modes involving real activated sludge microorganisms (24 h experiment) as well as duckweed plant (16 day experiment). Quantitative data of BB were obtained using microfluidic type device based on micro-TLC plate. This approach enabled direct determination of target component without sample pre-treatment like pre-concentration or pre-purification. Within single analytical run calibration line, retention standard spots (methyl red) and multiple samples were analyzed simultaneously. Due to the multivariate nature of these experiments, quantitative data were explored with chemometric tools including AHC (agglomerative hierarchical clustering), PCA (principal component analysis), and FA (factor analysis). Experimental data and multivariate calculations revealed that BB is strongly resistant on biodegradation, however, inclusion complexes formation with ß-cyclodextrinmay induce degradation of this dye in the presence of duckweed. It is hoped that results of our experimental work can be used for designing of future experiments for fast screening of different additives and improvement of technological processes, focusing on purification of sewage and water from micropollutants.

Carbon-Based Nanomaterials as Promising Material for Wastewater Treatment Processes.

In the latest literature search, the technology based on graphite oxide (GO) nanomaterials exhibits a great potential in many aspects of wastewater treatment involving adsorption, photocatalysis, disinfection and membrane process. In this study experimental data involving the carbon element in different forms such as active carbon (AC), graphite and graphene oxide (GO) applied as the active reagents in wastewater treatment are summarized and discussed. The first step was to characterize the aforementioned carbon materials and nanoparticles using various complementary techniques. These include optical microscopy, scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), Raman spectrophotometry and zeta potential measurements. The second issue was to design the relatively simple experiment enabling us to observe the physicochemical and biological effects of carbon nanoparticles in the presence of sewage water and/or active sludge. Obtained experimental data have been inspected using univariate and multivariate (principal component analysis, PCA) approaches confirming the complex interaction of GO nanoparticles with microorganisms that are present in activated sludge. This experiment enabled the collection of an initial data set to design different large scale investigations focusing on active nanoparticles affecting wastewater purification. PCA calculations clearly revealed that GO strongly affects the wastewater technological processes investigated. It is hoped that the described results will allow the design of smart environmental protection systems in the future.

Degradation Studies of Selected Bisphenols in the Presence of ß-Cyclodextrin and/or Duckweed Water Plant.

BACKGROUND: This research reports a multivariate experiment enabling observation of the potential application of macrocyclic compound [ß-cyclodextrin (ß-CD)] and/or duckweed organisms as the active factors for elimination of selected bisphenols A, B, and S from water samples. OBJECTIVE: Target bisphenols selection was based on observation that such components can be present in food or environmental samples (e.g., vegetable/fruit juices, milk, drinking water, or treated wastewater). METHODS: Biological research was carried out using aquatic organisms containing chlorophyll, particularly duckweed (Lemna minor L), that may work as an active biomass for the elimination or extraction of bisphenols micropollutants from water. Using such a system, we studied the potential encapsulation effect and removal efficiency of nontoxic macrocyclic oligosaccharide (ß-cyclodextrin) acting as an encapsulation reagent to promote the removal of selected bisphenols from liquid phase both with and without the presence of duckweed biomass. RESULTS: Experimental data have revealed that ß-CD or combined ß-CD/duckweed system has an effect on bisphenols elimination from water. The initial data set obtained from this preliminary experiment (and combined with supramolecular complex formation data calculated from chromatographic experiments, published previously) enables designing of further experiments focusing on the development of green chemistry technology. CONCLUSIONS: It is hoped that this may be used for the efficient removal of low-molecular-mass micropollutants using classical technological wastewater treatment processes modified by biomass and macrocyclic additives. This process needs to be optimized, but the results presented have revealed that such green chemistry technology, if successful, may be an interesting alternative for the selective removal of the micropollutants investigated from wastewater using classical adsorbents (e.g., carbons and carbon-related nanomaterials), particularly in terms of the worldwide problem with microplastic pollutants in the environment and food products.

Smart Sampling and Probing: Are You Getting All the Relevant Information?

BACKGROUND: Sampling (collecting) and probing (testing, measuring) are very common tasks in the analytical field, where we need to characterize a given system and complex samples. In this action, we try to ensemble maximal information related with the system under a given study, and, frequently, we may end an inefficient analytical situation. OBJECTIVE: The best way to avoid "oversampling" and "overprobing" is to evaluate the number of factors and objects that may be present in a current data set. METHODS: Suggested methodology in data analysis is mainly related with principal component analysis and principal object analysis. All used simulations and other controlled situations were here used to demonstrate how to retrieve the number of factors and objects present in a given data set and allow to supervise all sampling and probing process. RESULTS AND CONCLUSIONS: In this work, we explain and suggest how to use eigenvalue decomposition to access the actual number of factors and object contributions. A large pool of datasets were tested in order to assess the number of relevant features present in each dataset. HIGHLIGHTS: Proposed numerical approach was designed to supervise and help in sampling and probing process for the efficient analysis of complex systems such as those involving food and environmental samples.

Pilbara Craton Soil as A Possible Lunar Soil Simulant for Civil Engineering Applications.

Recent fast development in lunar exploration exposed a lack of lunar soil simulant (LSS) fit for civil engineering applications. Permanent human presence on the Moon will be associated with significant construction efforts. Adequate technologies and building materials have to be developed and tested prior to setting the actual building site on the Moon. Current LSSs were created for non-civil engineering purposes, thus they are very expensive and available in limited amounts. In the paper, the authors proved that Pilbara Craton soil is a suitable material for the creation of an affordable LSS for civil engineering applications. The main tool of the conducted study was principal component analysis (PCA).

Simple horizontal chamber for thermostated micro-thin-layer chromatography.

The aim of presented work is to describe simple, fast and robust temperature-controlled system for non-forced-flow micro-planar chromatography. With this separation system the micro-TLC plates can be developed in horizontal position under temperature gradient or non-gradient as well as saturated or unsaturated chamber conditions using low amount of mobile phase ranging from 0.3 to 1.0 mL. The device may work at wide range of temperatures from -20 to 80 degrees C. Under such conditions the plate temperature equilibration can be obtained within 5-12 min and a typical non-forced flow run can be finished within short period of time ranging from 5 to 20 min. It has been revealed that micro-plate is capable to separate more than 10 spots in one direction or up to 180 spots per plate for two-dimensional and multi-development runs. Particularly, fast and efficient separation of number of analytes including fullerenes, cyclodextrins and steroids as well as complex samples obtained from natural products and pharmaceutical formulations was demonstrated. Moreover, the application of thermostated micro-planar chromatography for the retention and quantification studies is also discussed.

Application of temperature-controlled micro planar chromatography for separation and quantification of testosterone and its derivatives.

Temperature-controlled micro thin-layer chromatography (TLC) was applied for separation and quantification studies of testosterone and its derivatives including methyltestosterone, testosterone propionate, isobutyrate, phenylpropionate, isocaproate, enanthate and caprate. Chromatographic studies were performed on silica-, octadecylsilica- and aluminum-coated plates working inside a small thermostated horizontal chamber unit allowing one-dimensional and two-dimensional developing modes with an elution distance of 45 mm. Retention properties of steroids were investigated across a whole range of binary mixtures such as methanol/water, acetonitrile/water, methanol/dichloromethane and acetone/hexane (0-100% v/v). Moreover, the effect of temperature ranging from -20 to +60 degrees C under saturated and unsaturated chamber conditions was also investigated. Our results revealed that depending on the mobile phase polarity the separation system based on the low carbon load wettable with water RP18W plates may work as a normal-phase (NP) or reversed-phase (RP) chromatographic system. It has been also demonstrated that micro TLC equipment can be applied as a fast retention screening device as well as simple and robust quantitative tool for determination of testosterone residue containing testosterone derivatives in complex samples.

Evaluation of the water and organic liquids extraction efficiency of Spirulina maxima dyes using thermostated micro thin-layer chromatography.

Thermostated micro thin-layer chromatography was applied for separation and quantification studies of Spirulina maxima dyes isolated from pharmaceutical formulation by a simple one-step liquid extraction. The isolation process was performed using a number of liquids, including water; 10 mM water solutions of native alpha-, beta-, and gamma-cyclodextrin and their hydroxypropyl derivatives; and a number of common organic liquids characterized by different polarity, namely, methanol, ethanol, 1-propanol, 2-propanol, acetonitrile, acetone, tetrahydrofuran, dichloromethane, toluene, and n-hexane. Chromatographic studies were performed on RP18W plates working inside a small thermostated horizontal chamber allowing a development distance of 45 mm. Using a mobile phase consisting of acetone-n-hexane (30 + 70, v/v) and 40 degrees C separation temperature, plate peak capacity of at least 15 spots/lane and developing time <5 min were obtained. Validation data indicated that under such conditions, with an office scanner used for chromatogram digitalization, spot quantification could be accurately performed within an analyte mass range of 2 factors. The raw quantitative data obtained from microchromatograms acquired under visible light conditions were explored using cluster analysis and principal components analysis. Chemometric investigations revealed that the best extraction liquids for isolation of dye mixtures from Spirulina samples were methanol, ethanol, tetrahydrofuran, and dichloromethane. Moreover, it was found that the liquids' parachor values could be used for estimation of the dye extraction efficiency from complex samples.

Dye Removal from Water and Wastewater Using Various Physical, Chemical, and Biological Processes.

Synthetic dyes or colorants are key chemicals for various industries producing textiles, food, cosmetics, pharmaceutics, printer inks, leather, and plastics. Nowadays, the textile industry is the major consumer of dyes. The mass of synthetic colorants used by this industry is estimated at the level of 1 ÷ 3 × 105 tons, in comparison with the total annual consumption of around 7 × 105 tons worldwide. Synthetic dyes are relatively easy to detect but difficult to eliminate from wastewater and surface water ecosystems because of their aromatic chemical structure. It should be highlighted that the relatively high stability of synthetic dyes leads to health and ecological concerns due to their toxic, mutagenic, and carcinogenic nature. Currently, removal of such chemicals from wastewater involves various techniques, including flocculation/coagulation, precipitation, photocatalytic degradation, biological oxidation, ion exchange, adsorption, and membrane filtration. In this review, a number of classical and modern technologies for synthetic dye removal from industry-originated wastewater were summarized and discussed. There is an increasing interest in the application of waste organic materials (e.g., compounds extracted from orange bagasse, fungus biosorbent, or green algal biomasses) as effective, low-cost, and ecologically friendly sorbents. Moreover, a number of dye removal processes are based on newly discovered carbon nanomaterials (carbon nanotubes and graphene as well as their derivatives).

Preliminary Studies of Synthetic Dye Adsorption on Iron Sludge and Activated Carbons.

There is great interest in the search for multifunctional waste-based materials that may be applied as environmentally friendly adsorbents. Iron-rich sludge from ground drinking-water treatment plants may be considered a potential adsorbent for various water contaminants. This material is generated during ground water purification because of the excess of metal ions in water (Fe, Mn). In practice, this sludge is frequently disposed of as waste material and, so far, is not commonly applied as the adsorption base. Our research aims to explore the adsorption potential of iron sludge for selected synthetic dyes, including malachite green, ponceau 4R, and brilliant blue FCF. Experimental data were performed using iron sludge collected from the Groundwater Treatment Plant in Koszalin, Poland, and comparing it with adsorption properties of commercial activated carbons (Norit SA Super and Norit CA 1). The kinetics, adsorption isotherms, and temperature influence on the removal of target dyes were investigated and discussed. Preliminary experimental data have revealed that iron sludge can be considered an adsorbent for the removal of cationic dyes.

Toward the Understanding of Micro-TLC Behavior of Various Dyes on Silica and Cellulose Stationary Phases Using A Data Mining Approach.

Planar chromatography and related techniques [micro-planar chromatography, micro-TLC, or paper-based microfluidic devices (µPADs)] present several advantages in analytical applications, such as simplicity, low cost of analysis, and the ability to work with raw complex samples without the involvement of time-consuming prepurification steps. By using commonly applied planar chromatographic systems and µPADs devices, stationary phases (silica and cellulose based), different solvent mixtures (methanol-water and dichloromethane-methanol), and proportions varying from 0 to 100% (v/v), micro-TLC migration profiles of several dyes described in terms of characteristic of chromatographic parameters (retardation factor, peak base width, and asymmetry factor) were investigated. Combining these results with some quantum mechanics calculated properties for each solute (dipole moment, polarizability), and by using the data mining approach, we modeled this overall chromatographic behavior in order to describe experimental data. With this approach, we were able to predict with reasonable confidence some chromatographic properties. This effort its crucial in order to (1) optimize solute elution, (2) increase mixture resolution, and (3) identify some molecular properties of analytes for designing simple micro-TLC. It is hoped that the presented nonhypothesis-driven data-mining approach can be helpful for understanding the chromatographic behavior of dyes on silica and cellulose adsorbents using the simplest mobile phases. This should be helpful for further designing the micro-TLC separation systems or µPADs quantification devices based on cellulose and related biopolymers and considering dye compounds as analytes for separation and sensing molecules.

Optimization of a solid phase extraction procedure for prostaglandin E2, F2 alpha and their tissue metabolites.

The primary prostaglandins PGE(2) and PGF(2 alpha) are metabolized in tissues by a series of enzymatic and non-enzymatic reactions. To measure metabolic rates and individual reaction rates it is necessary to extract the parent prostaglandins and metabolites before the separation and quantification of each compound is achieved. Here we have established and optimized a solid phase extraction (SPE) procedure to recover PGE(2), PGF(2 alpha) and their six enzymatic and non-enzymatic tissue metabolites from aqueous solutions including urine, plasma and tissue homogenate. We have used octadecyl-bonded silica gel as the stationary phase and methanol-water mixtures as binary mobile phases. The volumes and concentrations of the washing and elution solutions were optimized individually for each PG. Recoveries of all PG standards were quantitative except for PGEM, which was recovered at 80% efficiency. Biological matrix components interfered with the extraction in a PG- and matrix-specific fashion. Inclusion of 1% formic acid in the loading mixture raised recoveries from urine, plasma and tissue homogenate to >or=90%. This SPE method is the first that has been optimized by systematic elution studies for PGE(2), PGF(2 alpha) and the complement of their tissue metabolites. The procedure is simple, robust and can serve as an effective pre-purification step before downstream separation and quantification of each tissue metabolite of PGE(2) and PGF(2 alpha) from complex biological matrices.

Characterization of human fetal cord blood steroid profiles in relation to fetal sex and mode of delivery using temperature-dependent inclusion chromatography and principal component analysis (PCA).

In the present work, human male and female fetal cord blood samples were purified, selectively extracted and separated to examine a fraction of steroids ranging from polar estetrol to relatively non-polar progesterone using solid phase extraction based on C-18 tubes and beta-cyclodextrin driven temperature dependent inclusion chromatography. Resulting UV diode array chromatographic patterns revealed the presence of 27 peaks. Chromatographic patterns of UV detected steroids were analyzed using principal components analysis which revealed differences between male/female and labour/not-in-labour clusters. Quantitative analysis of nine identified steroids including: estetrol, 17beta-estradiol, estrone, estriol, cortisol, cortisone, progesterone, 20 alpha-hydroxyprogesterone and 17 alpha-hydroxyprogesterone were not significantly different between males and females. Significant differences between male and female fetuses were related to as yet unidentified compounds. Four peaks were significantly different with labour which corresponded with cortisol, cortisone and two unidentified compounds. This protocol may distinguish significant differences between clinical groups that are not readily identifiable using univariate measurements of single steroids or different low molecular mass biomarkers. Moreover, we have provided new evidence that despite the absence of testosterone there are number of steroids and low molecular mass compounds that differ between male and female fetuses.