Spot-test identification and rapid quantitative sequential analysis of dipyrone

A qualitative spot-test and tandem quantitative analysis of dipyrone in the bulk drug and in pharmaceutical preparations is proposed. The formation of a reddish-violet color indicates a positive result. In sequence a quantitative procedure can be performed in the same flask. The quantitative results obtained were statistically compared with those obtained with the method indicated by the Brazilian Pharmacopoeia, using the Student's t and the F tests. Considering the concentration in a 100 µL aliquot, the qualitative visual limit of detection is about 5×10-6 g; instrumental LOD ≅ 1.4×10-4 mol L-1 ; LOQ ≅ 4.5×10-4 mol L-1.

Cutaneous lesions sensory impairment recovery and nerve regeneration in leprosy patients

It is important to understand the mechanisms that enable peripheral neurons to regenerate after nerve injury in order to identify methods of improving this regeneration. Therefore, we studied nerve regeneration and sensory impairment recovery in the cutaneous lesions of leprosy patients (LPs) before and after treatment with multidrug therapy (MDT). The skin lesion sensory test results were compared to the histopathological and immunohistochemical protein gene product (PGP) 9.5 and the p75 nerve growth factor receptors (NGFr) findings. The cutaneous neural occupation ratio (CNOR) was evaluated for both neural markers. Thermal and pain sensations were the most frequently affected functions at the first visit and the most frequently recovered functions after MDT. The presence of a high cutaneous nerve damage index did not prevent the recovery of any type of sensory function. The CNOR was calculated for each biopsy, according to the presence of PGP and NGFr-immunostained fibres and it was not significantly different before or after the MDT. We observed a variable influence of MDT in the recovery from sensory impairment in the cutaneous lesions of LPs. Nociception and cold thermosensation were the most recovered sensations. The recovery of sensation in the skin lesions appeared to be associated with subsiding inflammation rather than with the regenerative activity of nerve fibres.

An electronic pressure-meter nociception paw test for mice

The aim of the present investigation was to describe and validate an electronic mechanical test for quantification of the intensity of inflammatory nociception in mice. The electronic pressure-meter test consists of inducing the animal hindpaw flexion reflex by poking the plantar region with a polypropylene pipette tip adapted to a hand-held force transducer. This method was compared to the classical von Frey filaments test in which pressure intensity is automatically recorded after the nociceptive hindpaw flexion reflex. The electronic pressure-meter and the von Frey filaments were used to detect time versus treatment interactions of carrageenin-induced hypernociception. In two separate experiments, the electronic pressure-meter was more sensitive than the von Frey filaments for the detection of the increase in nociception (hypernociception) induced by small doses of carrageenin (30 µg). The electronic pressure-meter detected the antinociceptive effect of non-steroidal drugs in a dose-dependent manner. Indomethacin administered intraperitoneally (1.8-15 mg/kg) or intraplantarly (30-300 µg/paw) prevented the hypersensitive effect of carrageenin (100 µg/paw). The electronic pressure-meter also detected the hypernociceptive effect of prostaglandin E2 (PGE2; 10-100 ng) in a dose-dependent manner. The hypernociceptive effect of PGE2 (100 ng) was blocked by dipyrone (160 and 320 µg/paw) but not by intraplantar administration of indomethacin (300 µg/paw). The present results validate the use of the electronic pressure-meter as more sensitive than the von Frey filaments in mice. Furthermore, it is an objective and quantitative nociceptive test for the evaluation of the peripheral antinociceptive effect of anti-inflammatory analgesic drugs, which inhibit prostaglandin synthesis (indomethacin) or directly block the ongoing hypernociception (dipyrone).

Determination of hydrochlorothiazide in pharmaceutical formulations by diffuse reflectance spectroscopy

This paper describes a method for quantitative spot test analysis of hydrochlorothiazide using diffuse reflectance spectroscopy. The reflectance measurements were performed analyzing the colored compound (l = 585 nm) produced from the reaction between hydrochlorothiazide and p-dimethylaminocinnamaldehyde (PDAC) in acid medium. This reaction occurred on filter paper after heating to 80ºC for 8 minutes. Factorial designs allowed varying multiple reaction factors simultaneously in order to obtain the best reaction conditions. These factors included heating temperature, heating time, acid volume and PDAC volume. The linearity was studied in the range of 3.36x10-2 to 1.01x10-1 mol L-1 with a correlation coefficient of 0.998. The limit of detection was estimated to be 1.32x10² mol L-1. Commercial samples were analyzed using the proposed method and the results were favorably compared with those of the United States Pharmacopeia method, showing that quantitative spot test analysis by diffuse reflectance could be successfully used to determine hydrochlorothiazide in medicines.

Quantitative toxoplasma gondii oocyst detection by a modified Kato Katz test using Kinyoun staining (KKK) in ME49 strain experimentally infected cats

We detected Toxoplasma gondii oocysts in feces of experimentally infected cats, using a Kato Katz approach with subsequent Kinyoun staining. Animals serologically negative to T. gondii were infected orally with 5x10² mice brain cysts of ME49 strain. Feces were collected daily from the 3rd to the 30th day after challenge. Oocysts were detected by qualitative sugar flotation and the quantitative modified Kato Katz stained by Kinyoun (KKK). In the experimentally infected cats, oocysts were detected from the 7th to 15th day through sugar flotation technique, but oocysts were found in KKK from the 6th to 16th day, being sensitive for a larger period, with permanent documentation. The peak of oocysts excretion occurred between the 8th to 11th days after challenge, before any serological positive result. KKK could be used in the screening and quantification of oocysts excretion in feces of suspected animals, with reduced handling of infective material, decreasing the possibility of environmental and operator contamination.

Sensory characteristics of 'Douradão' peaches submitted to modified atmosphere packaging

The sensory, physical and chemical characteristics of 'Douradão' peaches cold stored in different modified atmosphere packaging (LDPE bags of 30, 50, 60, 75µm thickness) were studied. After 14, 21 and 28 days of cold storage (1 ± 1 ºC and 90 ± 5% RH), samples were withdrawn from MAP and kept during 4 days in ambient air for ripening. Descriptive terminology and sensory profile of the peaches were developed by methodology based on the Quantitative Descriptive Analysis (QDA). The assessors consensually defined the sensory descriptors, their respective reference materials and the descriptive evaluation ballot. Fourteen individuals were selected as judges based on their discrimination capacity and reproducibility. Seven descriptors were generated showing similarities and differences among the samples. The data were analysed by ANOVA, Tukey test and Principal Component Analysis (PCA). The atmospheres that developed inside the different packaging materials during cold storage differed significantly. The PCA showed that MA50 and MA60 treatments were more characterized by the fresh peach flavour, fresh appearance, juiciness and flesh firmness, and were effective for keeping good quality of 'Douradão' peaches during 28 d of cold storage. The Control and MA30 treatments were characterized by the mealiness, the MA75 treatment showed lower intensity for all attributes evaluated and they were ineffective to maintain good quality of the fruits during cold storage. Higher correlation coefficients (positive) were found between fresh appearance and flesh firmness (0.95), fresh appearance and juiciness (0.97), ratio and intensity of fresh peach smell (0.81), as well as higher correlation coefficients (negative) between Hue angle and intensity of yellow colour (-0.91), fresh appearance and mealiness (-0.92), juiciness and mealiness (-0.95), firmness and mealiness (-0.94).

Sensory profiles of chocolates produced from cocoa cultivars resistant to Moniliophtora Perniciosa

The present study evaluated the sensory quality of chocolates obtained from two cocoa cultivars (PH16 and SR162) resistant to Moniliophtora perniciosa mould comparing to a conventional cocoa that is not resistant to the disease. The acceptability of the chocolates was assessed and the promising cultivars with relevant sensory and commercial attributes could be indicated to cocoa producers and chocolate manufacturers. The descriptive terminology and the sensory profile of chocolates were developed by Quantitative Descriptive Analysis (QDA). Ten panelists, selected on the basis of their discriminatory capacity and reproducibility, defined eleven sensory descriptors, their respective reference materials and the descriptive evaluation ballot. The data were analyzed using ANOVA, Principal Component Analysis (PCA) and Tukey's test to compare the means. The results revealed significant differences among the sensory profiles of the chocolates. Chocolates from the PH16 cultivar were characterized by a darker brown color, more intense flavor and odor of chocolate, bitterness and a firmer texture, which are important sensory and commercial attributes. Chocolates from the SR162 cultivar were characterized by a greater sweetness and melting quality and chocolates from the conventional treatment presented intermediate sensory characteristics between those of the other two chocolates. All samples indicated high acceptance, but chocolates from the PH16 and conventional cultivars obtained higher purchase intention scores.

Terbinafine: optimization of a LC method for quantitative analysis in pharmaceutical formulations and its application for a tablet dissolution test

A simple liquid chromatographic method was optimized for the quantitative determination of terbinafine in pharmaceutical hydroalcoholic solutions and tablets, and was also employed for a tablet dissolution test. The analysis was carried out using a RP-C18 (250 mm × 4.6 mm, 5 μm) Vertical® column, UV-Vis detection at 254 nm, and a methanol-water (95:5, v/v) mobile phase at a flow-rate of 1.2 mL min-1. Method validation investigated parameters such as linearity, precision, accuracy, robustness and specificity, which gave results within the acceptable range. The tablets dissolution was quite fast: 80% of the drug was dissolved within 15 min.

Sensory profile of Italian salami with coriander (Coriandrum sativum L.) essential oil

The descriptive terminology and sensory prolife of four samples of Italian salami were determined using a methodology based on the Quantitative Descriptive Analysis (QDA). A sensory panel consensually defined sensory descriptors, their respective reference materials, and the descriptive evaluation ballot. Twelve individuals were selected as judges and properly trained. They used the following criteria: discriminating power, reproducibility, and individual consensus. Twelve descriptors were determined showing similarities and differences among the Italian salami samples. Each descriptor was evaluated using a 10 cm non-structured scale. The data were analyzed by ANOVA, Tukey test, and the Principal Component Analysis (PCA). The salami with coriander essential oil (T3) had lower rancid taste and rancid odor, whereas the control (T1) showed high values of these sensory attributes. Regarding brightness, T4 showed the best result. For the other attributes, T1, T2, T3, and T4 were similar.

Sensory profile of 'Douradão' peaches cold stored under controlled atmosphere

The sensory quality of 'Douradão' peaches cold stored in three different conditions of controlled atmosphere (CA1, CA2, CA3 and Control) was studied. After 14, 21 and 28 days of cold storage, samples were withdrawn from CA and kept for 4 days in ambient air for ripening. The sensory profile of the peaches and the descriptive terminology were developed by methodology based on the Quantitative Descriptive Analysis (QDA). The panelists consensually defined the sensory descriptors, their respective reference materials and the descriptive evaluation ballot. Fourteen panelists were selected based on their discrimination capacity and reproducibility. Seven descriptors were generated showing similarities and differences between samples. The data were analyzed by ANOVA, Tukey test and Principal Component Analysis (PCA). Results showed significant differences in the sensory profiles of the peaches. The PCA showed that CA2 and CA3 treatments were more characterized by the fresh peach flavor, fresh peach appearance, juiciness and flesh firmness, and were effective in keeping the good quality of the 'Douradão' peaches during the 28 days of cold storage. The Control and CA1 treatments were characterized by the mealiness and were ineffective for quality maintenance of the fruits during cold storage.

Sensory profile of eleven peach cultivars

The goal of this study was to evaluate the sensory profile of eleven peach cultivars grown in an experimental orchard located in the city of Lapa (PR, Brazil) in two seasons. The peach cultivars analyzed were Aurora I, Chimarrita, Chiripá, Coral, Eldorado, Granada, Leonense, Maciel, Marli, Premier, and Vanguarda. The sensory analysis was performed by previously trained panelists; 20 of them in the first season and 10 in the second season. The sensory evaluation was performed using Quantitative Descriptive Analysis, in which the following attributes were measured: appearance, aroma, flesh color, flesh firmness, flavor, and juiciness. The results showed preference for sweet, soft, and juicy fruits. Chimarrita, Chiripá, and Coral fruits showed better sensorial performance than the other peach cultivars. It was also verified that the analysis of the attributes aroma, flesh firmness, and flavor is enough for performing the sensory profile of peach fruits for in natura consumption.

Sensory profile and drivers of liking for grape nectar among smoker and nonsmoker consumers

Decreased gustatory and olfactory capacity is one of the problems caused by tobacco use. The objectives of this study were to determine the sensory profile of six grape nectar samples sweetened with different sweeteners and to verify the drivers of liking in two distinct consumer groups: smokers and nonsmokers. The sensory profile was constructed by twelve trained panelists using quantitative descriptive analysis (QDA). Consumer tests were performed with 112 smokers and 112 nonsmokers. Partial least squares regression analyses was used to identify the drivers of acceptance and rejection of the grape nectars among the two consumer groups. According to the QDA, the samples differed regarding six of the nineteen attributes generated. The absolute averages of the affective test were lower in the group of smokers; possibly because smoking influences acceptance and eating preferences, especially with regard to sweet foods. The results showed that the grape flavor was the major driver of preference for acceptance of the nectar, while astringency, wine aroma, bitterness and sweetness, and bitter aftertaste were drivers of rejection in the two groups of consumers, with some differences between the groups.

Microimmunodiffusion test for the serodiagnosis of paracoccidioidomycosis

We used the micro- and macroimmunodiffusion test for the qualitative and quantitative measurement of anti - P. brasiliensis antibodies in serum of patients with paracoccidioidomycosis. All 103 paracoccidioidomycosis sera (100%) were positive in the micro test versus 87% positivity index in the macrotest. All 83 control sera from patients with other diseases were negative in both tests. Titers of the positive sera tended to be higher in the microtest, which revealed sharper and easier to read precipiting bands. Microimmunodiffusion is simple to be performed, requires a minimum amount of reagents and allows the simultaneous testing of 102 sera. It may replace the macrotest specially in laboratories dealing with great serologic routine.

Comparison on the performance of Leishmania major-like and Leishmania braziliensis braziliensis as antigen for new world leishmaniasis IgG-immunofluorescence test

The performance of an antigen of L. major-like promastigotes for the serological diagnosis of mucocutaneous leishmaniasis in the IgG-immunofluorescent test was compared to that of an antigen of L. braziliensis braziliensis. Each antigen was used to test two hundred and twenty-four sera of etiologies such as mucocutaneous leishmaniasis, deep mycoses, toxoplasmosis, malaria, Chagas' disease, visceral leishmaniasis, anti-nuclear factor, schistosomaiasis, rheumatoid factor and normal controls. Agreement between responses to each antigen was high: 77.2% of leishmaniases sera agreed on a positive or a negative result to both antigens and 91.1 % of control sera. Cross reactivity was restricted to Chagas' disease sera, visceral leishmaniasis, anti-nuclear factor and paracoccidiodomycosis. The quantitative response of leishmaniasis and Chagas' disease sera to both antigens was evaluated by a linear regression; although the y-intercept and the slope were different for each antigen, neither was better than the other in the disclosure of anti-Leishmania antibodies. In the case of Chagas' disease sera the L. major-like antigen was better than L. b. braziliensis' to disclose cross-reacting antibodies.

A low-cost method to test cytotoxic effects of Crotalus vegrandis (Serpentes: Viperidae) venom on kidney cell cultures

The pathogenesis of the renal lesion upon envenomation by snakebite has been related to myolysis, hemolysis, hypotension and/or direct venom nephrotoxicity caused by the venom. Both primary and continuous cell culture systems provide an in vitro alternative for quantitative evaluation of the toxicity of snake venoms. Crude Crotalus vegrandis venom was fractionated by molecular exclusion chromatography. The toxicity of C. vegrandis crude venom, hemorrhagic, and neurotoxic fractions were evaluated on mouse primary renal cells and a continuous cell line of Vero cells maintained in vitro. Cells were isolated from murine renal cortex and were grown in 96 well plates with Dulbecco's Modified Essential Medium (DMEM) and challenged with crude and venom fractions. The murine renal cortex cells exhibited epithelial morphology and the majority showed smooth muscle actin determined by immune-staining. The cytotoxicity was evaluated by the tetrazolium colorimetric method. Cell viability was less for crude venom, followed by the hemorrhagic and neurotoxic fractions with a CT50 of 4.93, 18.41 and 50.22 µg/mL, respectively. The Vero cell cultures seemed to be more sensitive with a CT50 of 2.9 and 1.4 µg/mL for crude venom and the hemorrhagic peak, respectively. The results of this study show the potential of using cell culture system to evaluate venom toxicity.