CE methods for analysis of isoforms of prostate-specific antigen compatible with online derivatization for LIF detection

Prostate-specific antigen (PSA) is the usual biomarker for prostate cancer (PCa). However, its lack of selectivity has lead to the search for new biomarkers. PSA glycosylation seems to depend on the pathophysiological conditions of the individual. Thus, methods to separate PSA isoforms (peaks) to study their role as PCa markers are needed. In this work, CE methods for PSA isoforms separation, based on the use of different dynamic coatings, are developed using UV detection. Three complementary CE methods allowing the separation of 8 or 9 PSA isoforms are selected. The longest method takes only 17?min, while the shortest one separates 9 isoforms in < 8?min. Depending on the isoforms of interest for their use as PCa biomarker, the CE method to be used can be chosen or various of them can be combined. A remarkable aspect of these methods is that the BGEs employed are devoid of compounds with primary amino groups, making the CE methods compatible with fluorescent on-column derivatization through amino residues. As a proof-of-concept, a preliminary result shows that LIF detection of labeled PSA analyzed by one of the three developed methods permits detection of glycoprotein isoforms.     

Effect of synthesis conditions on the microstructure of TEOS derived silica hydrogels synthesized by the alcohol-free sol–gel route

Silica matrices synthesized from a pre-hydrolysis step in ethanol followed by alcohol removal at low pressure distillation, and condensation in water, are suitable for encapsulation of biomolecules and microorganisms and building bioactive materials with optimized optical properties. Here we analyze the microstructure of these hydrogels from the dependence of I(q) data acquired from SAXS experiments over a wide range of silica concentration and pH employed in the condensation step. From the resulting data it is shown that there is a clear correlation between the microscopic parameters—cluster fractal dimension (D), elementary particle radius (a) and cluster gyration radius (R)—with the attenuation of visible light when the condensation step proceeds at pH < 6. At higher pHs, there is a steep dependence of the cluster density (~R ^D−3) with the condensation pH, and non-monotonous changes of attenuance are less than 20%, revealing the complexity of the system. These results, which were obtained for a wide pH and silica concentration range, reinforce the idea that the behavior of gels determined in a restricted interval of synthesis variables cannot be extrapolated, and comparison of gelation times is not enough for predicting their properties.     

Centrifugally-driven sample extraction, preconcentration and purification in microfluidic compact discs

Centrifugally-driven microfluidic compact discs (μ-CDs) have attracted significant interest within the analytical science community in the past decade, with the primary focus on the potential of such platforms for performing parallel and/or multiplex biological assays and further application in biomedical diagnostics. More recently, μ-CD-based devices were also applied to environmental analysis as platforms for multi-sample extraction and transportation, prior to off-disc analysis in the laboratory. This review critically summarizes recent developments in μ-CD platforms for sample extraction, preconcentration, fractionation and purification in bioanalytical and environmental applications. We also summarize the common methods employed in the fabrication of μ-CD platforms. Further, we discuss preparation of stationary phases in microfluidic channels embedded in μ-CDs, as applications of μ-CDs in sample extraction are generally based on enclosed series of extraction phases and microcolumns.     

Analytical techniques in the study of highly-nitrated nitrocellulose

This work presents an updated overview of the analytical techniques used to study highly-nitrated nitrocellulose, which is used in explosives and is of forensic interest. Most articles published in the past decade were designed:(1) to investigate polymeric parameters of nitrocellulose (e.g., molar mass distribution, viscosity and specific refractive index) by size-exclusion chromatography;(2) to determine the morphological and thermal characteristics of nitrocellulose using thermal and spectroscopic techniques; and,(3) to study the thermal, biological and mechanical degradation of nitrocellulose by thermal, spectroscopic, and mass spectrometric (MS) techniques, alone or coupled to gas chromatography.However, the few papers that focused on the determination of nitrocellulose used in explosives employed analytical techniques [e.g., vibrational techniques (infrared and Raman spectroscopy), MS and ion-mobility spectrometry (IMS) and liquid chromatography (LC) (high-performance LC and ion chromatography)]. Most of the information reported by these techniques has been qualitative. Only quantitative determination of nitrocellulose or its nitrogen content has been performed by measuring the nitrite and/or nitrate ions released from its basic hydrolysis.     

Electromembrane extraction (EME) and HPLC determination of non-steroidal anti-inflammatory drugs (NSAIDs) in wastewater samples

In this paper, an electromembrane extraction (EME) combined with a HPLC procedure using diode array (DAD) and fluorescence detection (FLD) has been developed for the determination of six widely used non-steroidal anti-inflammatory drugs (NSAIDs): salicylic acid (SAC), ketorolac (KTR), ketoprofen (KTP), naproxen (NAX), diclofenac (DIC) and ibuprofen (IBU). The drugs were extracted from basic aqueous sample solutions, through a supported liquid membrane (SLM) consisting of 1-octanol impregnated in the walls of a S6/2 Accurel^® polypropylene hollow fiber, and into a basic aqueous acceptor solution resent inside the lumen of the hollow fiber with a potential difference of 10 V applied over the SLM. Extractions that were carried out in 10 min using a potential of 10 V from pH 12 NaOH aqueous solutions shown concentration enrichments factors of 28-49 in a pH 12 NaOH aqueous acceptor solution. The proposed method was successfully applied to urban wastewaters. Excellent selectivity was demonstrated as no interfering peaks were detected. The procedure allows very low detection and quantitation limits of 0.0009-9.0 and 0.003-11.1 μg L⁻¹, respectively.     

Study of the capillary electrophoresis profile of intact α-1-acid glycoprotein isoforms as a biomarker of atherothrombosis

α-1-Acid glycoprotein (AGP) is a serum glycoprotein that presents several isoforms. Changes in the isoforms of AGP have been related to different pathological states including cardiovascular diseases (CVDs) such as acute myocardial infarction. However, to our knowledge, the role of variations of AGP isoforms as a potential biomarker of atherothrombosis has not been addressed. In this work, a preliminary study about differences in the capillary zone electrophoresis (CZE) profile of intact (non-hydrolyzed) AGP isoforms between healthy individuals and patients with atherothrombosis, specifically abdominal aortic aneurysm (AAA) and carotid atherosclerosis (CTA), has been performed. Biological samples (plasmas and sera) were analyzed by CZE after immunoaffinity chromatography purification. Up to 13 peaks corresponding to groups of isoforms of intact AGP from plasma samples were detected by CZE-UV. Electrophoretic profiles were aligned, peaks assigned, and linear discriminant analysis (LDA) of percentage of the corrected areas of AGP peaks was employed to discriminate and classify the CZE profiles of AGP samples. LDA enabled to accomplish 92.9% of correct classification of the AGP samples when the three groups of samples were considered. Besides, the LDA model showed high predictive power in the groups healthy vs. sick, healthy vs. AAA, and healthy vs. CTA. The described method was a successful approach to study the potential of AGP isoforms profile as a biomarker of atherothrombosis. To the best of our knowledge this has been the first time that a possible role of the CZE profile of intact AGP isoforms as a biomarker of vascular diseases has been demonstrated.     

Highly stereoselective double (R)-phenylglycinol-induced cyclocondensation reactions of symmetric aryl bis(oxoacids)

The double cyclocondensation of symmetric pyridyl bis(oxoacids) 2b and 3b with (R)-phenylglycinol stereoselectively gave access to bis-phenylglycinol-derived oxazolopyrrolidine 9 and oxazolopiperidone 10, respectively. Application of the stereocontrolled cyclocondensation reaction to phenyl bis-γ-oxoacid 4b provided 11, which was converted to the corresponding enantiopure di(pyrrolidinyl)benzene 22. The absolute configuration of the new stereogenic centers generated in the key cyclocondenstion step was unambiguously established by X-ray crystallographic analysis.     

Synthesis of trifluoromethyl cyclohexyl, cyclohexenyl and aryl compounds via stepwise Robinson annulation

The diketone 2-fluoro-2-(trifluoromethyl)-1-phenylhexane-1,5-dione 3 was synthesized by a Mukaiyama Michael type reaction from the corresponding tetrafluoroenol silyl ether prepared from pentafluoropropiophenone. This diketone was treated under basic conditions and was converted, depending on the stoichiometry of the base, into the surprisingly stable ketol 4-fluoro-4-(trifluoromethyl)-3-hydroxy-3-phenylcyclohexanone 4 as a single diastereomer (catalytic KOH) or to the biphenylol 6-(trifluoromethyl)biphenyl-3-ol (excess KOH, THF) 5. Solvolysis of the trifluoromethyl group (anionic activation) occurred using excess KOH in alcohol. The corresponding cyclohexenone derivative 7, the usual product of Robinson annulation, might be prepared in good yield via mesylation of the ketol. Thus various unprecedented fluorinated cyclohexane and aromatic derivatives were achieved in a few steps from the commercially available pentafluoropropiophenone.     

Intracrystalline diffusion in metal organic framework during heterogeneous catalysis: influence of particle size on the activity of MIL-100 (Fe) for oxidation reactions

Three MIL-100 (Fe) samples differing in average crystal size (from 60-70 to >400 nm) have been synthesized by microwave heating using three HF/Fe(3+) ratios. Oxidation of diphenylmethane with tert-butylhydroperoxide (TBHP) and thiophenol with oxygen are catalyzed by three MIL-100 (Fe) samples with similar reaction rates regardless of its average particle size. In contrast, the activity of the three MIL-100 (Fe) samples for the oxidation of bulky triphenylmethane by TBHP largely depends on the average crystal size of the sample: the smaller the average particle size, the larger the initial reaction rate of triphenylmethane oxidation. These results show that diffusion limitation takes place on MOF catalysis depending on the substrate size and provides indirect evidence that these reactions take place inside the intracrystalline space of the porous catalysts.     

Determination of ethylene glycol dinitrate in dynamites using HPLC: application to the plastic explosive Goma-2 ECO

In this study, a sequential extraction method using water and methanol to recover ethylene glycol dinitrate or nitroglycol (EGDN) contained in Goma-2 ECO dynamite was developed. After, an HPLC method was used for the determination of EGDN in the two extracted phases. The analytical method was validated by evaluating its selectivity, sensitivity, linearity, and linear working concentration range, limit of detection and quantitation, precision (as repeatability and intermediate precision), accuracy, and robustness, providing appropriate values (i.e. RSD values for precision about 6% and accuracy about 100%). Finally, the EGDN content of a sample of the Goma-2-ECO dynamite was determined obtaining a concentration of 30.29%, which is in accordance with the manufacturer's specifications for this dynamite (25.7-31.4%).