129 resultados para SOLID-PHASE
Resumo:
In the present study we determined the efficacy of the measurement of fecal cortisol and androgen metabolite concentrations to monitor adrenal and testicular activity in the jaguar (Panthera onca). Three captive male jaguars were chemically restrained and electroejaculated once or twice within a period of two months. Fecal samples were collected daily for 5 days before and 5 days after the procedure and stored at -20ºC until extraction. Variations in the concentrations of cortisol and androgen metabolites before and after the procedure were determined by solid phase cortisol and testosterone radioimmunoassay and feces dry weight was determined by drying at 37ºC for 24 h under vacuum. On four occasions, fecal cortisol metabolite levels were elevated above baseline (307.8 ± 17.5 ng/g dry feces) in the first fecal sample collected after the procedure (100 to 350% above baseline). On one occasion, we did not detect any variation. Mean (± SEM) fecal androgen concentration did not change after chemical restraint and electroejaculation (before: 131.1 ± 26.7, after: 213.7 ± 43.6 ng/g dry feces). These data show that determination of fecal cortisol and androgen metabolites can be very useful for a noninvasive assessment of animal well-being and as a complement to behavioral, physiological, and pathological studies. It can also be useful for the study of the relationship between adrenal activity and reproductive performance in the jaguar.
Resumo:
Alternative methods to the utilization of laboratory animal blood and its by-products are particularly attractive, especially regarding hamsters due to their small size and difficulties in obtaining serial blood samples. Steroid hormone metabolite quantification in feces, widely used in studies of free-ranging or intractable animals, is a non-invasive, non-stressor, economical, and animal saving technique which allows longitudinal studies by permitting frequent sampling of the same individual. The present study was undertaken to determine the suitability of this method for laboratory animals. Estradiol and progesterone metabolites were quantified by radioimmunoassay in feces of intact, sexually mature female Syrian hamsters during the estrous cycle (control) and in feces of superovulated females. Metabolites were extracted by fecal dilution in ethanol and quantified by solid phase radioimmunoassay. Median estrogen and progesterone concentrations were 9.703 and 180.74 ng/g feces in the control group, respectively. Peaks of estrogen (22.44 ± 4.54 ng/g feces) and progesterone (655.95 ± 129.93 ng/g feces) mean fecal concentrations respectively occurred 12 h before and immediately after ovulation, which is easily detected in this species by observation of a characteristic vaginal postovulatory discharge. Median estrogen and progesterone concentrations (28.159 and 586.57 ng/g feces, respectively) were significantly higher in superovulated animal feces (P < 0.0001). The present study demonstrated that it is possible to monitor ovarian activity in Syrian hamsters non-invasively by measuring fecal estradiol and progesterone metabolites. This technique appears to be a quite encouraging method for the development of new endocrinologic studies on laboratory animals.
Resumo:
The purpose of the present study was to compare the sensitivity and specificity of V3 enzyme immunoassay (solid phase EIA and EIA inhibition) and restriction fragment length polymorphism (RFLP) with the DNA sequencing "gold standard" to identify the Brazilian HIV-1 variants of subtype B and B"-GWGR. Peripheral blood mononuclear cells were collected from 61 HIV-1-infected individuals attending a clinic in São Paulo. Proviral DNA was amplified and sequentially cleaved with the Fok I restriction enzyme. Plasma samples were submitted to a V3-loop biotinylated synthetic peptide EIA. Direct partial DNA sequencing of the env gene was performed on all samples. Based on EIA results, the sensitivity for detecting B-GPGR was 70%, compared to 64% for the Brazilian variant B"-GWGR while, the specificity of B-GPGR detection was 85%, compared to 88% for GWGR. The assessment of RFLP revealed 68% sensitivity and 94% specificity for the B-GPGR strain compared to 84 and 90% for the B"-GWGR variant. Moreover, direct DNA sequencing was able to detect different base sequences corresponding to amino acid sequences at the tip of the V3 loop in 22 patients. These results show a similar performance of V3 serology and RLFP in identifying the Brazilian variant GWGR. However, V3 peptide serology may give indeterminate results. Therefore, we suggest that V3 serology be used instead of DNA sequencing where resources are limited. Samples giving indeterminate results by V3 peptide serology should be analyzed by direct DNA sequencing to distinguish between B-GPGR and the Brazilian variant B"-GWGR.
Resumo:
The volatile components of noni at two ripening stages were isolated by headspace solid-phase microextraction using 65 µm Polydimethylsiloxane-Divinylbenzene (PDMS/DVB) fibers and analyzed using GC/MS. Both maturation stages had several compounds in common. Ninety-six compounds were identified, from which octanoic acid ( 70% of total extract) and hexanoic acid ( 8% of total extract) were found to be the major constituents. Due to noni maturation, octanoic acid, decanoic acid and 2E-nonenal decreased their concentrations, while some esters (methyl hexanoate, methyl octanoate, ethyl octanoate and methyl 4E-decenoate), which their fruity odor notes, increased their contents. Two unsaturated esters, reported for the first time in this fruit, 3-methyl-3-buten-1-yl hexanoate and 3-methyl-3-buten-1-yl octanoate, significantly decreased their concentration in the ripe to over-ripe fruits.
Resumo:
Simultaneous Distillation-Extraction (SDE) and headspace-solid phase microextraction (HS-SPME) combined with GC-FID and GC-MS were used to analyze volatile compounds from plum (Prunus domestica L. cv. Horvin) and to estimate the most odor-active compounds by application of the Odor Activity Values (OAV). The analyses led to the identification of 148 components, including 58 esters, 23 terpenoids, 14 aldehydes, 11 alcohols, 10 ketones, 9 alkanes, 7 acids, 4 lactones, 3 phenols, and other 9 compounds of different structures. According to the results of SDE-GC-MS, SPME-GC-MS and OAV, ethyl 2-methylbutanoate, hexyl acetate, (E)-2-nonenal, ethyl butanoate, (E)-2-decenal, ethyl hexanoate, nonanal, decanal, (E)-β-ionone, Γ-dodecalactone, (Z)-3-hexenyl acetate, pentyl acetate, linalool, Γ-decalactone, butyl acetate, limonene, propyl acetate, Δ-decalactone, diethyl sulfide, (E)-2-hexenyl acetate, ethyl heptanoate, (Z)-3-hexenol, (Z)-3-hexenyl hexanoate, eugenol, (E)-2-hexenal, ethyl pentanoate, hexyl 2-methylbutanoate, isopentyl hexanoate, 1-hexanol, Γ-nonalactone, myrcene, octyl acetate, phenylacetaldehyde, 1-butanol, isobutyl acetate, (E)-2-heptenal, octadecanal, and nerol are characteristic odor active compounds in fresh plums since they showed concentrations far above their odor thresholds.
Resumo:
Existing data about the aroma of fresh-cut watermelon and the metabolic changes that occur with minimal processing are scarce. Given the close relationship that exists between aroma, texture, and quality characteristics, it is necessary to investigate the changes in the volatile profile and texture of watermelon, a fruit extensively sold in supermarket chains throughout Brazil. The objective of this work was to analyze the volatile profile using solid phase microextraction (SPME) as well as texture changes in fresh-cut watermelon stored at 5 °C for ten days. Chromatography associated with sensory analysis (sniffing) led us to conclude that 9-carbon (C9) alcohols and aldehydes are the major responsible for the flavor and aroma of minimally processed watermelon stored at 5 ± 1 °C/90 ± 5% RH for ten days, and also that the aroma diminishes in intensity with storage, but it does not affect the final quality of the product. It was noted that the amount of drained liquid, soluble pectin, and weight loss increased during storage concurrently with a reduction in firmness and a structural breakdown of the cells. Pectin methyl esterase activity remained constant and polygalacturonase activity was not detected.
Resumo:
Increased preference for healthy and functional foods could be an opportunity to increase the consumption of clarified cashew apple juice. Given its level of fructose, glucose, and vitamin C, it can be used as a base in blends. However, its characteristic odor can interfere with the acceptance of these formulations, especially by consumers who are not familiar with cashew aroma. The aim of this study was to evaluate the effect of treatment with macroporous resins (FPA54, FPX66, XAD761, and XAD4) on the volatile profile and physicochemical characteristics of clarified cashew apple juice. After the treatment with the resins, the volatile profile was evaluated using solid-phase microextraction (SPME) and gas chromatography/mass spectrometry (GC/MS). The physicochemical analyses performed were: pH, soluble solids (ºBrix), total titrable acidity, reducing sugars, and vitamin C. Gas chromatography analyses showed that XAD4 and FPX66 led to a reduction of the initial amount of volatile compounds to 14.05% and 15.72%, respectively. These two resins also did not affect the physicochemical characteristics of the clarified cashew apple juice.
Resumo:
The aim of this study was to extract and identify volatile compounds from pineapple residues generated during concentrated juice processing. Distillates of pineapple residues were obtained using the following techniques: simple hydrodistillation and hydrodistillation by passing nitrogen gas. The volatile compounds present in the distillates were captured by the solid-phase microextraction technique. The volatile compounds were identified in a system of high resolution gas chromatography system coupled with mass spectrometry using a polyethylene glycol polar capillary column as stationary phase. The pineapple residues constituted mostly of esters (35%), followed by ketones (26%), alcohols (18%), aldehydes (9%), acids (3%) and other compounds (9%). Odor-active volatile compounds were mainly identified in the distillate obtained using hydrodistillation by passing nitrogen gas, namely decanal, ethyl octanoate, acetic acid, 1-hexanol, and ketones such as γ-hexalactone, γ-octalactone, δ-octalactone, γ-decalactone, and γ-dodecalactone. This suggests that the use of an inert gas and lower temperatures helped maintain higher amounts of flavor compounds. These data indicate that pineapple processing residue contained important volatile compounds which can be extracted and used as aroma enhancing products and have high potential for the production of value-added natural essences.
Resumo:
Abstract Brazilian wine production is characterized by Vitis labrusca grape varieties, especially the economically important Isabel cultivar, with over 80% of its production destined for table wine production. The objective of this study was to optimize and validate the conditions for extracting volatile compounds from wine with the solid-phase microextraction technique, using the response surface method. Based on the response surface analysis, it can be concluded that the central point values maximize the process of extracting volatile compounds from wine, i.e., an equilibrium time of 15 minutes, an extraction time of 35 minutes, and an extraction temperature of 30 °C. Esters were the most numerous compounds found under these extraction conditions, indicating that wines made from Isabel cultivar grapes are characterized by compounds that confer a fruity aroma; this finding corroborates the scientific literature.