Accelerated Solvent Extraction for Gas Chromatographic Analysis of Nicotine and Cotinine in Meconium Samples

Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)

Partitioning and extraction of collagenase from Penicillium aurantiogriseum in poly(ethylene glycol)/phosphate aqueous two-phase system

Purification of collagenase produced by Penicillium aurantiogriseum URM4622 was carried using a PEG/phosphate aqueous two-phase system (ATPS). A 2(3)-full experimental design was used to investigate the influence of PEG molar mass, PEG concentration and phosphate concentration on the selected responses, namely partition coefficient, activity yield and purification factor. The ATPS was composed of PEG (molar mass of 550, 1500 and 4000 g/mol) at concentrations of 15.0, 17.5 and 20.0% (w/w) and phosphate at concentrations of 12.5, 15.0 and 17.5% (w/w). The best results of one-step extraction of collagenase from the fermentation broth (partition coefficient of 1.01, activity yield of 242% and purification factor of 23.5) were obtained at pH 6.0 using 20.0% (w/w) PEG 550 and 17.5% (w/w) phosphate. The results of this preliminary study demonstrate that the selected ATPS is satisfactorily selective for the extraction of such a collagenase. (C) 2012 Elsevier B.V. All rights reserved.

Effects of the extraction conditions on the yield and composition of rice bran oil extracted with ethanol-A response surface approach

Rice bran oil was obtained from rice bran by solvent extraction using ethanol. The influence of process variables, solvent hydration (0-24% of water, on mass basis), temperature (60-90 degrees C), solvent-to-rice bran mass ratio (2.5:1 to 4.5:1) and stirrer speed (100-250 rpm) were analysed using the response surface methodology. The extraction yield was highly affected by the solvent water content, and it varied from 8.56 to 20.05 g of oil/100 g of fresh rice bran (or 42.7-99.9% of the total oil available) depending on the experimental conditions. It was observed that oryzanol and tocols behave in different ways during the extraction process. A larger amount of tocols is extracted from the solid matrix in relation to gamma-oryzanol. It was possible to obtain values from 123 to 271 mg of tocols/kg of fresh rice bran and 1527 to 4164 mg of oryzanol/kg of fresh rice bran, indicating that it is feasible to obtain enriched oil when this renewable solvent is used. No differences in the chemical composition of the extracted oils were observed when compared to the data cited in the literature. (C) 2011 The Institution of Chemical Engineers. Published by Elsevier B.V. All rights reserved.

A simple boiling-based DNA extraction for RAPD profiling of landfarm soil to provide representative metagenomic content

Landfarm soils are employed in industrial and petrochemical residue bioremediation. This process induces selective pressure directed towards microorganisms capable of degrading toxic compounds. Detailed description of taxa in these environments is difficult due to a lack of knowledge of culture conditions required for unknown microorganisms. A metagenomic approach permits identification of organisms without the need for culture. However, a DNA extraction step is first required, which can bias taxonomic representativeness and interfere with cloning steps by extracting interference substances. We developed a simplified DNA extraction procedure coupled with metagenomic DNA amplification in an effort to overcome these limitations. The amplified sequences were used to generate a metagenomic data set and the taxonomic and functional representativeness were evaluated in comparison with a data set built with DNA extracted by conventional methods. The simplified and optimized method of RAPD to access metagenomic information provides better representativeness of the taxonomical and metabolic aspects of the environmental samples.

Liquid-liquid equilibria for systems composed of rice bran oil and alcohol-rich solvents: Application to extraction and deacidification of oil

The liquid-liquid equilibria of systems composed of rice bran oil, free fatty acids, ethanol and water were investigated at temperatures ranging from 10 to 60 degrees C. The results of the present study indicated that the mutual solubility of the compounds decreased with an increase in the water content of the solvent and a decrease in the temperature of the solution. The experimental data set was correlated by applying the UNIQUAC model. The average variance between the experimental and calculated compositions was 0.35%, indicating that the model can accurately predict behavior of the compounds at different temperatures and degrees of hydration. The adjustment of interaction parameters enables both the simulation of liquid-liquid extractors for deacidification of vegetable oil and the prediction of phase compositions for the oil and alcohol-rich phases that are generated during cooling of the stream exiting the extractor (when using ethanol as the solvent). (C) 2012 Elsevier Ltd. All rights reserved.

Pre- and Postoperative Quantitative Analysis of Contour Abnormalities in Graves Upper Eyelid Retraction

Purpose: One of the most common problems of the surgical management of Graves upper eyelid retraction is the occurrence of eyelid contour abnormalities. In the present study, the postoperative contour of a large sample of eyelids of patients with Graves orbitopathy was measured. Methods: The postoperative upper eyelid contour of 62 eyes of 43 patients with Graves orbitopathy was subjectively classified by 3 experienced surgeons in 3 categories: poor, fair, and good. The shape of the eyelid contours in each category was then measured with a recently developed custom-made software by measuring multiple midpupil eyelid distances each 15 degrees along the palpebral fissure. The upper eyelid contour of 60 normal subjects was also quantified as a control group. Results: The mean ratio between the sum of the lateral and medial midpupil eyelid distances (lateral/medial ratio) was 1.10 +/- 0.11 standard deviation in controls and 1.15 +/- 0.13 standard deviation in patients. Postoperatively, the mean midpupil eyelid distance at 90 degrees was 4.16 +/- 1.13 mm standard deviation. The distribution lateral/medial ratios of the eyelids judged as having good contours was similar to the distribution of the controls with a modal value centered on the interval between 1.0 and 1.10. The distribution of lateral/medial ratios of the eyelids judged as having poor contour was bimodal, with eyelids with low and high lateral/medial ratios. Low lateral/medial ratios occurred when there was a lateral overcorrection, giving the eyelid a flat or a medial ptosis appearance. High lateral/medial ratios were due to a central or medial overcorrection or a lateral peak maintenance. Conclusions: Postoperative upper eyelid contour abnormalities can be quantified by comparing the sum of multiple midpupil eyelid distances of the lateral and medial sectors of the eyelid. Low and high lateral/medial ratios are anomalous and judged as unpleasant. (Ophthal Plast Reconstr Surg 2012;28:429-433)

Simultaneous Quantification of Lycopene, beta-Carotene, Retinol and alpha-Tocopherol in Plasma after a Simple Extraction Procedure: Stability Study and Application to Human Volunteers

A method for the simultaneous quantification of lycopene, beta-carotene, retinol and alpha-tocopherol by high-performance liquid chromatography (HPLC) with Vis/fluorescence detection with isocratic elution was optimized and validated. The method consists of a rapid and simple liquid-liquid extraction procedure and a posterior quantification of extracted supernatants by HPLC. Aliquots of plasma were stored at -20 degrees C for three months for stability study. The methodology was applied to samples from painters and individuals not exposed to paints (n = 75). The assay was linear for all vitamins (r > 0.99). Intra-and inter-run precisions were obtained with coefficient of variation smaller than 5%. The accuracies ranged from 0.29 to -5.80% and recoveries between 92.73 and 101.97%. Plasma samples and extracted supernatants were stable for 60 days at -20 degrees C. A significant decrease of lycopene, beta-carotene and retinol concentrations in plasma from exposed individuals compared to non-exposed individuals (p < 0.05) was observed. The method is simple, reproducible, precise, accurate and sensitive, and can be routinely utilized in clinical laboratories.

An environmentally friendly analytical procedure for nickel determination by atomic and molecular spectrometry after cloud point extraction in different samples

Cloud point extraction (CPE) was employed for separation and preconcentration prior to the determination of nickel by graphite furnace atomic absorption spectrometry (GFAAS), flame atomic absorption spectrometry (FAAS) or UV-Vis spectrophotometry. Di-2-pyridyl ketone salicyloylhydrazone (DPKSH) was used for the first time as a complexing agent in CPE. The nickel complex was extracted from the aqueous phase using the Triton X-114 surfactant. Under optimized conditions, limits of detection obtained with GFAAS, FAAS and UV-Vis spectrophotometry were 0.14, 0.76 and 1.5 mu g L-1, respectively. The extraction was quantitative and the enrichment factor was estimated to be 27. The method was applied to natural waters, hemodialysis concentrates, urine and honey samples. Accuracy was evaluated by analysis of the NIST 1643e Water standard reference material.

Fractionation of lemon essential oil by solvent extraction: Phase equilibrium for model systems at T=298.2 K

The fractioning of lemon essential oil can be performed by liquid-liquid extraction using hydrous ethanol as a solvent. A quaternary mixture composed of limonene, gamma-terpinene, beta-pinene, and citral was used to simulate lemon essential oil. In this paper, we present (liquid + liquid) equilibrium data that were experimentally determined for systems containing essential oil compounds, ethanol, and water at T = 298.2 K. The experimental data were correlated using the NRTL and UNIQUAC models, and the mean deviations between calculated and experimental data were less than 0.0053 in all systems, indicating the accuracy of these molecular models in describing our systems. The results show that as the water content in the solvent phase increased, the values of the distribution coefficients decreased, regardless of the type of compound studied. However, the oxygenated compound always showed the highest distribution coefficient among the components of the essential oil, thus making deterpenation of the lemon essential oil a feasible process. (C) 2012 Elsevier Ltd. All rights reserved.

Assessment of Preemptive Analgesia Efficacy in Surgical Extraction of Third Molars

Background and objectives: Literature on preemptive analgesia is controversial. Reliability of results and difficult reproducibility of research contribute for non-elucidation of the subject. The aim of this study is to test the efficacy of oral ketoprofen (150 mg) preemptively administrated two days before third molar surgery, compared with postoperative administration in the same patient. Methods: Thirteen patients underwent surgical removal of bilateral third molar in two separate procedures. In a random and double blind procedure, oral ketoprofen 150 mg was administered every 12 hours two days before surgery and, after the procedure, the same drug was administered for three days. On the other side, a control (placebo) was used orally every 12 hours two days before surgery and, after the procedure, ketoprofen 150 mg was administered every 12 hours for three days. Postoperative pain was assessed by visual analogue scale, nominal scale, and amount of rescue analgesics consumed. Results: There was no statistically significant difference in postoperative pain between the preemptive treatment and control. Conclusion: In this experimental model, preemptive analgesia was not effective in reducing postoperative pain in surgical extraction of third molar compared with the postoperative administration of the same drug.

Perception of the esthetic impact of mandibular incisor extraction treatment on laypersons, dental professionals, and dental students

Objective: To evaluate the degree of perception of laypersons, dental professionals, and dental students regarding dental esthetics in cases with mandibular central incisor extraction. Materials and Methods: Using a smile photograph of a person with normal occlusion and all teeth, modifications were made to reflect the extraction of a mandibular incisor of various compositions and sizes. For this purpose a program specifically for image manipulation (Adobe Photoshop CS3, Adobe Systems Inc) was used. After manipulation the images were printed on photographic paper, attached to a questionnaire and distributed to laypersons, dental professionals, and dental students (n = 90) to evaluate the degree of perception and esthetic using a scale of attractiveness, where 0 = hardly attractive, 5 = attractive, and 10 = very attractive. The differences between examiners were checked by the Mann-Whitney test. All the statistics were performed with a confidence level of 95%. Results: The results demonstrated the skill of the dental professionals and dental students in perceiving the difference between cases of normal occlusion and cases where an incisor was lacking (P < .05). The photograph in which the lateral incisors were shown to be larger than the central incisor was the one that obtained the highest value among the cases of extraction in all groups of evaluators. Conclusions: It can be concluded that dental professionals and dental students are more skillful at identifying deviation from normality. In addition, central incisor extraction should always be discarded when there are other treatment options available. (Angle Orthod. 2012;82:732-738.)

Multiple Radial Midpupil Lid Distances: A Simple Method for Lid Contour Analysis

Purpose: To describe a new computerized method for the analysis of lid contour based on the measurement of multiple radial midpupil lid distances. Design: Evaluation of diagnostic technology. Participants and Controls: Monocular palpebral fissure images of 35 patients with Graves' upper eyelid retraction and of 30 normal subjects. Methods: Custom software was used to measure the conventional midpupil upper lid distance (MPLD) and 12 oblique MPLDs on each 15 degrees across the temporal (105 degrees, 120 degrees, 135 degrees, 150 degrees, 165 degrees, and 180 degrees) and nasal (75 degrees, 60 degrees, 45 degrees, 30 degrees, 15 degrees, and 0 degrees) sectors of the lid fissure. Main Outcome Measures: Mean, standard deviation, 5th and 95th percentiles of the oblique MPLDs obtained for patients and controls. Temporal/nasal MPLD ratios of the same angles with respect to the midline. Results: The MPLDs increased from the vertical midline in both nasal and temporal sectors of the fissure. In the control group the differences between the mean central MPLD (90 degrees) and those up to 30 degrees in the nasal (75 degrees and 60 degrees) and temporal sectors (105 degrees and 120 degrees) were not significant. For greater eccentricities, all temporal and nasal mean MPLDs increased significantly. When the MPLDs of the same angles were compared between groups, the mean values of the Graves' patients differed from control at all angles (F = 4192; P<0.0001). The greatest temporal/nasal asymmetry occurred 60 degrees from the vertical midline. Conclusions: The measurement of radial MPLD is a simple and effective way to characterize lid contour abnormalities. In patients with Graves' upper eyelid retraction, the method demonstrated that the maximum amplitude of the lateral lid flare sign occurred at 60 degrees from the vertical midline. Financial Disclosure(s): The authors have no proprietary or commercial interest in any of the materials discussed in this article. Ophthalmology 2012; 119: 625-628 (C) 2012 by the American Academy of Ophthalmology.

Sequential extraction of phosphorus in an Oxisol amended with biosolids in a long-term field experiment in Brazil

The continued growth of large cities is producing increasing volumes of urban sewage sludge. Disposing of this waste without damaging the environment requires careful management. The application of large quantities of biosolids (treated sewage sludge) to agricultural lands for many years may result in the excessive accumulation of nutrients like phosphorus (P) and thereby raise risks of eutrophication in nearby water bodies. We evaluated the fractionation of P in samples of an Oxisol collected as part of a field experiment in which biosolids were added at three rates to a maize (Zea mays L) plantation over four consecutive years. The biosolids treatments were equivalent to one, two and four times the recommended N rate for maize crops. In a fourth treatment, mineral fertilizer was applied at the rate recommended for maize. Inorganic P forms were extracted with ammonium chloride to remove soluble and loosely bound P; P bound to aluminum oxide (P-Al) was extracted with ammonium fluoride; P bound to iron oxide (P-Fe) was extracted with sodium hydroxide; and P bound to calcium (P-Ca) was extracted with sulfuric acid. Organic P was calculated as the difference between total P and inorganic P. The predominant fraction of P was P-Fe, followed by P-Al and P-Ca. P fractions were positively correlated to the amounts of P applied, except for P-Ca. The low values of P-Ca were due to the advanced weathering processes to which the Oxisol have been subjected, under which forms of P-Ca are converted to P-Fe and P-Al. The fertilization with P via biosolids increased P availability for maize plants even when a large portion of P was converted to more stable forms. Phosphorus content in maize leaves and grains was positively correlated with P fractions in soils. From these results it can be concluded that the application of biosolids in highly weathered tropical clayey soils for many years, even above the recommended rate based on N requirements for maize, tend to be less potentially hazardous to the environment than in less weathered sandy soils because the non-readily P fractions are predominant after the addition of biosolids. (C) 2012 Elsevier B.V. All rights reserved.

Analysis of Insect Cuticular Compounds by Non-lethal Solid Phase Micro Extraction with Styrene-Divinylbenzene Copolymers

Insect cuticular hydrocarbons including relatively non-volatile chemicals play important roles in cuticle protection and chemical communication. The conventional procedures for extracting cuticular compounds from insects require toxic solvents, or non-destructive techniques that do not allow storage of subsequent samples, such as the use of SPME fibers. In this study, we describe and tested a non-lethal process for extracting cuticular hydrocarbons with styrene-divinylbenzene copolymers, and illustrate the method with two species of bees and one species of beetle. The results demonstrate that these compounds can be efficiently trapped by ChromosorbA (R) (SUPELCO) and that this method can be used as an alternative to existing methods.

Vaginal morcellation: A new strategy for large gynecological malignant tumor extraction A pilot study

Objective. Evaluate feasibility and safety of a novel technique for uterine morcellation in patients scheduled for laparoscopic treatment of gynecologic malignances. Background. The laparoscopic management of uterine malignancies is progressively gaining importance and popularity over laparotomy. Nevertheless, minimal invasive surgery is of limited use when patients have enlarged uterus or narrow vagina. In these cases, conventional uterus morcellation could be a solution but should not be recommended due to risks of tumor dissemination. Methods. Prospective pilot study of women with endometrial cancer in which uterus removal was a realistic concern due to both organ size and proportionality. Brief technique description: after completion of total laparoscopic hysterectomy and bilateral anexectomy, a nylon with polyurethane Lapsac (R) is vaginally inserted into the abdomen; the specimen is placed inside the pouch that will be closed and rotated 180 degrees toward the vaginal vault and, posteriorly, pushed into the vaginal canal; in the transvaginal phase, the surgeon pulls the edges of the bag up to vaginal introitus and all vaginal walls will be covered; inside the pouch, the operator performs a uterus bisection-morcellation. Results. In our series of 8 cases, we achieved successful completion in all patients, without conversion to laparotomy. Average operative time, blood loss and length of hospitalization were favorable. One patient presented with a vesicovaginal fistula. Conclusion. The vaginal morcellation following oncologic principles is a feasible method that permits a rapid uterine extraction and may avoid a number of unnecessary laparotomies. Further studies are needed to confirm the oncological safety of the technique. (C) 2012 Elsevier Inc. All rights reserved.