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1.
Anal Bioanal Chem ; 394(8): 2087-94, 2009 Aug.
Article in English | MEDLINE | ID: mdl-19575192

ABSTRACT

Designed experiments based on a simplex mixture design were employed to explore the effects of three solvent components (water, formic acid, and aqueous acetic acid), extraction time, and extraction temperature for the automated microextraction of basic (cationic) dyes from acrylic fibers. Extractions were conducted by an automated liquid handling system, and dye extraction was evaluated using a UV/visible microplate reader. Highest extraction efficiency for two subclasses of basic dyes (methine and azo) from acrylic fibers was achieved with an extraction solvent containing 88% formic acid/12% water. Cationic dyes were analyzed by capillary electrophoresis using a 45 mM ammonium acetate buffer in acetonitrile-water at pH 4.7. The utility of microextraction combined with capillary electrophoresis-mass spectrometry for analysis of extracts from trace fibers was demonstrated by the detection and characterization of three basic dyes extracted from a 2-mm length of single acrylic fiber.

2.
Anal Bioanal Chem ; 394(8): 2077-85, 2009 Aug.
Article in English | MEDLINE | ID: mdl-19543716

ABSTRACT

The extraction and separation of dyes present on textile fibers offers the possibility of enhanced discrimination between forensic trace fiber evidence. An automated liquid sample handling workstation was programmed to deliver varying solvent combinations to acid-dyed nylon samples, and the resulting extracts were analyzed by an ultraviolet/visible microplate reader to evaluate extraction efficiencies at different experimental conditions. Combinatorial experiments using three-component mixture designs varied three solvents (water, pyridine, and aqueous ammonia) and were employed at different extraction temperatures for various extraction durations. The extraction efficiency as a function of the three solvents (pyridine/ammonia/water) was modeled and used to define optimum conditions for the extraction of three subclasses of acid dyes (anthraquinone, azo, and metal complex) from nylon fibers. The capillary electrophoresis analysis of acid dye extracts is demonstrated using an electrolyte solution of 15 mM ammonium acetate in acetonitrile/water (40:60, v/v) at pH 9.3. Excellent separations and discriminating diode array spectra are obtained even for dyes of similar color.

3.
FEBS Lett ; 580(16): 3780-6, 2006 Jul 10.
Article in English | MEDLINE | ID: mdl-16780840

ABSTRACT

(1-->3)-alpha-glucanases catalyze the hydrolysis of fungal cell wall (1-->3)-alpha-glucan, and function during cell division of yeasts containing this cell wall component or act in mycoparasitic processes. Here, we characterize the mechanism of action of the (1-->3)-alpha-glucanase MutAp from the mycoparasitic fungus Trichoderma harzianum. We observed that MutAp releases predominantly beta-glucose upon hydrolysis of crystalline (1-->3)-alpha-glucan, indicating inversion of the anomeric configuration. After having identified (1-->3)-alpha-glucan tetrasaccharide as the minimal substrate for MutAp, we showed that reduced (1-->3)-alpha-glucan pentasaccharide is cleaved into a trisaccharide and a reduced disaccharide, demonstrating that MutAp displays endo-hydrolytic activity. We propose a model for the catalytic mechanism of MutAp, whereby the enzyme breaks an intrachain glycosidic linkage of (1-->3)-alpha-glucan, and then continues its hydrolysis towards the non-reducing end by releasing beta-glucose residues in a processive manner.


Subject(s)
Glycoside Hydrolases/metabolism , Trichoderma/enzymology , Carbohydrate Conformation , Catalysis , Glucans/chemistry , Glucans/metabolism , Glucose/metabolism , Glycoside Hydrolases/antagonists & inhibitors , Hydrolysis , Models, Biological , Polysaccharides/metabolism , Substrate Specificity
4.
Appl Spectrosc ; 59(8): 986-92, 2005 Aug.
Article in English | MEDLINE | ID: mdl-16105206

ABSTRACT

Nylons are an important class of synthetic polymers, from an industrial, as well as forensic, perspective. A spectroscopic method, such as Fourier transform infrared (FT-IR) spectroscopy, is necessary to determine the nylon subclasses (e. g., nylon 6 or nylon 6,6). Library searching using absolute difference and absolute derivative difference algorithms gives inconsistent results for identifying nylon subclasses. The objective of this study was to evaluate the usefulness of peak ratio analysis and multivariate statistics for the identification of nylon subclasses using attenuated total reflection (ATR) spectral data. Many nylon subclasses could not be distinguished by the peak ratio of the N-H vibrational stretch to the sp(3) C-H(2) vibrational stretch intensities. Linear discriminant analysis, however, provided a graphical visualization of differences between nylon subclasses and was able to correctly classify a set of 270 spectra from eight different subclasses with 98.5% cross-validated accuracy.

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