Mass spectrometric analysis of glucose-modified ribonuclease

RNase A (1 mM) was incubated with glucose (0.4 M) at 37°C for up to 14 days in phosphate buffer (0.2 M, pH 7.4), digested with trypsin and analysed by LC-MS. The major sites of fructoselysine formation were Lys1, Lys7, Lys37 and Lys41. Three of these sites (Lys7, Lys37 and Lys41) were also the major sites of Ne-(carboxymethyl)lysine formation.

Flavor characterization of sugar-added pennywort (Centella asiatica L.) juices treated with ultra-high pressure and thermal processes

The flavor characteristics of pennywort juices with added sugar treated by ultra-high pressure, pasteurization, and sterilization were investigated using solid phase microextraction combined with gas chromatography-mass spectrometry. It was found that sesquiterpene hydrocarbons comprised the major class of volatile components present and the juices had a characteristic aroma due to the presence of volatiles including beta-caryophyllene and humulene and alpha-copaene. In comparison with heated juices, HPP-treated samples could retain more volatile compounds such as linalool and geraniol similar to those present in fresh juice, whereas some volatiles such as alpha-terpinene and ketone class were apparently formed by thermal treatment. All processing operations produced juice that was not significantly different in the concentration of total volatiles. Practical Application: Pennywort juice is considered a nutraceutical drink for health benefits. Therefore, to preserve all aroma and active components in this juice, a nonthermal process such as ultra-high pressure should be a more appropriate technique for retention of its nutritive values than pasteurization and sterilization.

Rifaximin modulates the colonic microbiota of patients with Crohn's disease: an in vitro approach using a continuous culture colonic model system.

Rifaximin, a rifamycin derivative, has been reported to induce clinical remission of active Crohn's disease (CD), a chronic inflammatory bowel disorder. In order to understand how rifaximin affects the colonic microbiota and its metabolism, an in vitro human colonic model system was used in this study. We investigated the impact of the administration of 1800 mg/day of rifaximin on the faecal microbiota of four patients affected by colonic active CD [Crohn's disease activity index (CDAI > 200)] using a continuous culture colonic model system. We studied the effect of rifaximin on the human gut microbiota using fluorescence in situ hybridization, quantitative PCR and PCR–denaturing gradient gel electrophoresis. Furthermore, we investigated the effect of the antibiotic on microbial metabolic profiles, using 1H-NMR and solid phase microextraction coupled with gas chromatography/mass spectrometry, and its potential genotoxicity and cytotoxicity, using Comet and growth curve assays. Rifaximin did not affect the overall composition of the gut microbiota, whereas it caused an increase in concentration of Bifidobacterium, Atopobium and Faecalibacterium prausnitzii. A shift in microbial metabolism was observed, as shown by increases in short-chain fatty acids, propanol, decanol, nonanone and aromatic organic compounds, and decreases in ethanol, methanol and glutamate. No genotoxicity or cytotoxicity was attributed to rifaximin, and conversely rifaximin was shown to have a chemopreventive role by protecting against hydrogen peroxide-induced DNA damage. We demonstrated that rifaximin, while not altering the overall structure of the human colonic microbiota, increased bifidobacteria and led to variation of metabolic profiles associated with potential beneficial effects on the host.

Flow injection analysis of ethyl xanthate by gas diffusion and UV detection as CS(2) for process monitoring of sulfide ore flotation

A sensitive and robust analytical method for spectrophotometric determination of ethyl xanthate, CH(3)CH(2)OCS(2)(-) at trace concentrations in pulp solutions from froth flotation process is proposed. The analytical method is based on the decomposition of ethyl xanthate. EtX(-), with 2.0 mol L(-1) HCl generating ethanol and carbon disulfide. CS(2). A gas diffusion cell assures that only the volatile compounds diffuse through a PTFE membrane towards an acceptor stream of deionized water, thus avoiding the interferences of non-volatile compounds and suspended particles. The CS(2) is selectively detected by UV absorbance at 206 nm (epsilon = 65,000 L mol(-1) cm(-1)). The measured absorbance is directly proportional to EtX(-) concentration present in the sample solutions. The Beer`s law is obeyed in a 1 x 10(-6) to 2 x 10(-4) mol L(-1) concentration range of ethyl xanthate in the pulp with an excellent correlation coefficient (r = 0.999) and a detection limit of 3.1 x 10(-7) mol L(-1), corresponding to 38 mu g L. At flow rates of 200 mu L min(-1) of the donor stream and 100 mu L min(-1) of the acceptor channel a sampling rate of 15 injections per hour could be achieved with RSD < 2.3% (n = 10, 300 mu L injections of 1 x 10(-5) mol L(-1) EtX(-)). Two practical applications demonstrate the versatility of the FIA method: (i) evaluation the free EtX(-) concentration during a laboratory study of the EtX(-) adsorption capacity on pulverized sulfide ore (pyrite) and (ii) monitoring of EtX(-) at different stages (from starting load to washing effluents) of a flotation pilot plant processing a Cu-Zn sulfide ore. (C) 2010 Elsevier By. All rights reserved.

Síntese e caracterização de catalisadores nanométricos de LaSrNiO4 para aplicação em dessulfurização de tiofeno

The mixed metal oxides constitute an important class of catalytic materials widely investigated in different fields of applications. Studies of rare earth nickelates have been carried by several researchers in order to investigate the structural stability afforded by oxide formed and the existence of catalytic properties at room temperature. So, this study aims synthesize the nanosized catalyst of nickelate of lanthanum doped with strontium (La(1-x)SrxNiO4-d; x = 0,2 and 0,3), through the Pechini method and your characterization for subsequent application in the desulfurization of thiophene reaction. The precursor solutions were calcined at 300ºC/2h for pyrolysis of polyester and later calcinations occurred at temperatures of 500 - 1000°C. The resulting powders were characterized by thermogravimetric analysis (TG / DTG), surface area for adsorption of N2 by BET method, X-ray diffraction (XRD), scanning electron microscopy (HR_SEM) and spectrometry dispersive energy (EDS). The results of XRD had show that the perovskites obtained consist of two phases (LSN and NiO) and from 700ºC have crystalline structure. The results of SEM evidenced the obtainment of nanometric powders. The results of BET show that the powders have surface area within the range used in catalysis (5-50m2/g). The characterization of active sites was performed by reaction of desulfurization of thiophene at room temperature and 200ºC, the relation F/W equal to 0,7 mol h-1mcat -1. The products of the reaction were separated by gas chromatography and identified by the selective detection PFPD sulfur. All samples had presented conversion above 95%

Comparison of the quantification of caffeine in human plasma by gas chromatography and ELISA

In the present study we evaluated the precision of the ELISA method to quantify caffeine in human plasma and compared the results with those obtained by gas chromatography. A total of 58 samples were analyzed by gas chromatography using a nitrogen-phosphorus detector and routine techniques. For the ELISA test, the samples were diluted to obtain a concentration corresponding to 50% of the absorbance of the standard curve. To determine whether the proximity between the I50 of the standard curve and that of the sample would bring about a more precise result, the samples were divided into three blocks according to the criterion of difference, in modulus, of the I50 of the standard curve and of the I50 of the sample. The samples were classified into three groups. The first was composed of 20 samples with I50 up to 1.5 ng/ml, the second consisted of 21 samples with I50 ranging from 1.51 to 3 ng/ml, and the third of 17 samples with I50 ranging from 3.01 to 13 ng/ml. The determination coefficient (R² = 0.999) showed that the data obtained by gas chromatography represented a reliable basis. The results obtained by ELISA were also reliable, with an estimated Pearson correlation coefficient of 0.82 between the two methods. This coefficient for the different groups (0.88, 0.79 and 0.49 for groups 1, 2 and 3, respectively) showed greater reliability for the test with dilutions closer to I50.

Desenvolvimento de uma unidade laboratorial para quantificação de BTX como poluentes atmosféricos, usando microextração em fase sólida

This dissertation aims the development of an experimental device to determine quantitatively the content of benzene, toluene and xylenes (BTX) in the atmosphere. BTX are extremely volatile solvents, and therefore play an important role in atmospheric chemistry, being precursors in the tropospheric ozone formation. In this work a BTX new standard gas was produced in nitrogen for stagnant systems. The aim of this dissertation is to develop a new method, simple and cheaper, to quantify and monitor BTX in air using solid phase microextraction/ gas chromatography/mass spectrometry (SPME/CG/MS). The features of the calibration method proposed are presented in this dissertation. SPME sampling was carried out under non-equilibrium conditions using a Carboxen/PDMS fiber exposed for 10 min standard gas mixtures. It is observed that the main parameters that affect the extraction process are sampling time and concentration. The results of the BTX multicomponent system studied have shown a linear and a nonlinear range. In the non-linear range, it is remarkable the effect of competition by selective adsorption with the following affinity order p-xylene > toluene > benzene. This behavior represents a limitation of the method, however being in accordance with the literature. Furthermore, this behavior does not prevent the application of the technique out of the non-linear region to quantify the BTX contents in the atmosphere.

Volatile compounds from organic and conventional passion fruit (Passiflora edulis F. Flavicarpa) pulp

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Coordination polymer adsorbent for matrix solid-phase dispersion extraction of pesticides during analysis of dehydrated Hyptis pectinata medicinal plant by GC/MS

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Identification of polycyclic aromatic hydrocarbons and methoxylated phenols in wood smoke emitted during production of charcoal

More than 130 organic substances in dichloromethane-methanol (4: 1) extracts of particulate matter and the gaseous phase from wood burning for the production of charcoal have been identified by capillary gas chromatography coupled with low-resolution mass spectrometry (GC-MS), use of GC retention indices, and comparison with authentic standards. Many of the substances identified are methoxyphenols (derivatives of syringol and guaiacol), polycyclic aromatic hydrocarbons (PAH), oxidized PAH (oxy-PAH), and levoglucosan, the last being a monosoccharide derivative from the thermal breakdown of cellulose. The amount of unsubstituted PAH was greater than that of methyl- and dimethyl-substituted homologs.

OFF-LINE SUPERCRITICAL-FLUID EXTRACTION - HIGH-RESOLUTION GAS-CHROMATOGRAPHY APPLIED TO THE STUDY OF MORACEAE SPECIES

Supercritical fluid extraction with CO2, Performed in a home-made system, of rhizomes of Dorstenia bryoniifolia Mart. ex Miq. (Moraceae) and of bark roots of Brosimum gaudichaudii Trecul (Moraceae) afforded crude extracts that were analysed by high resolution gas chromatography (HRGC). The D. bryoniifolia extract contained, besides the previously reported pimpinelin and isobergapten, the furocoumarins psoralen, bergapten, isopimpinelin and the triterpenes alpha- and beta-amyrin and the acetate of the latter. The B. gaudichaudii extract contained a number of terpenoids as well as the previously reported psoralen and bergapten. Supercritical fluid extraction gave extracts qualitatively similar to those obtained by Soxhlet extraction with hexane and, together with off-line HRGC, was shown to be a fast and accurate technique to be used in rapid phytochemical examination.

Screening Brazilian automotive gasoline quality through quantification of saturated hydrocarbons and anhydrous ethanol by gas chromatography and exploratory data analysis

In this paper a set of Brazilian commercial gasoline representative samples from São Paulo State, selected by HCA, plus six samples obtained directly from refineries were analysed by a high-sensitive gas chromatographic (GC) method ASTM D6733. The levels of saturated hydrocarbons and anhydrous ethanol obtained by GC were correlated with the quality obtained from Brazilian Government Petroleum, Natural Gas and Biofuels Agency (ANP) specifications through exploratory analysis (HCA and PCA). This correlation showed that the GC method, together with HCA and PCA, could be employed as a screening technique to determine compliance with the prescribed legal standards of Brazilian gasoline.

Determination of the sodium monofluoroacetate in serum by gas chromatography

Sodium monofluoroacetate (NAFAc) has been widely used for vertebrate pest control, such as rabbits in Australia. However NAFAc is extremely toxic to all vertebrates and its use is restricted. Although this compound is stringently restricted, the occurrence of accidental and homicidal poisoning is no ever-present possibility.The method developed in this work shows the applicability of SPE with alumina cartridges for the extraction of NAFAc from serum samples. The method is efficient with recoveries of at least 96.8% from spiked serum. The samples were subsequently derivatized with dicyclohexylcabodiimide (DCC), using 2,4-dichloroaniline (DCA), to make the product volatile for GC analysis.

Essential oil composition of Aloysia triphylla (L'Herit) Britton leaves cultivated in Botucatu, São Paulo, Brazil

The family Verbenaceae comprises about 175 genera and 2300 species, distributed in tropics and subtropics, mainly in temperate zone of southern hemisphere. The lemon verbena (Aloysia triphylla (L'Herit) Britton) is a perennial, bushy plant originally from South America. The essential oil of this plant is used in pharmaceutical, cosmetic and perfumery industry. Therapeutic properties include febrifuge, sedative, stomachical, diuretic, and antispasmodic activities. The present work aimed to identify the chemical composition of essential oil of Aloysia triphylla leaves. The study was done in Lageado Experimental Farm of the Department of Plant Production-Horticulture, Agronomical Sciences College, São Paulo State University Campus of Botucatu. Leaves of lemon verbena from Medicinal and Aromatic Plant Garden, were collected in the end of winter (September/2001). The essential oil was extracted by hydrodistillation, in Clevenger apparatus. 100 g of leaves were used in each extraction. Four extractions were performed during three hours. The essential oils of the leaves were analyzed in Gas Chromatography Mass spectrometry (CG-MS, Shimadzu, QP-5000), equipped with capillary column DB-5 (30 m × 0,25 mm × 0,25 mm), split 1/35, injector for 220 C°, detector for 230 C°, dragged by gas He (1,0 mL/min), with programmed temperature for 60 C° to 240 C°, 3 C°/min. The identification of the substances was held by comparison of their mass spectra with data of the CG-MS (Nist 62 lib), literature references and retention index of Kovats. The main constituents of essential oils were geranial (29.54 %), neral (27.01 %), limonene (15.93 %), geranyl acetate (4.0 %) and geraniol (3.96 %). This species possesses high quantity of monoterpenes and low quantity of sesquiterpenes.

Composition of coriander essential oil from Brazil

Coriander (Coriandrum sativum L.) is an annual and herbaceous plant, belonging to the Apiaceae family. Native of southern Europe and western Mediterranean region, this herb is cultivated world widely. This species, rich in linalool, has potential using as source of essential oil and as a medicinal plant. It has been used as analgesic, carminative, digestive, depurative, anti-rheumatic and antispasmodic agent. Its fruits (commonly called seeds) are used for flavoring candies, in cookery, perfumery, beverage and in tobacco industry. The aim of this study was to analyze the chemical composition of the seed essential oil of this species grown in Botucatu, São Paulo, Brazil. The experiment was carried out in Lageado Experimental Farm, Department of Plant Production, Agronomical Sciences College, São Paulo State University. The fruits were harvest 108 days after sowing. The essential oils were extracted by hydro distillation, in Clevenger apparatus. 50 g of fruits were used in each extraction. Three extractions were performed during three hours. The essential oils were analyzed in Gas Chromatography Mass Spectrometer (CG-MS, Shimadzu, QP-5000), equipped with DB-5 capillary column (30 m × 0,25 mm × 0,25 mm), split 1/20, injector for 240 C°, detector for 230 C°, dragged by gas He (1,7 mL/min), with programmed temperature for 40 C° (5 min)-150 C°, 4 C°/min; 150 C°-280 C°, 8 C°/min. The identification of the compounds was made by comparison of their spectra of masses with data from CG-MS (Nist 62 lib), literature references and retention index of Kovats. The 18 most important components were identified and quantified. The main components of the oil were linalool (77.48 %), γ-terpinene (4.64 %), α-pinene (3.97 %), limonene (1.28 %), geraniol (0.64 %) and 2-decenal (0.16 %).