Coeficientes de difusão de metais em materiais não convencionais (agarose e acetato de celulose) usados na técnica de difusão em filmes finos por gradientes de concentração

The DGT technique allows one to measure quantitatively free and labile metal species in aquatic systems. Nevertheless, for this approach, knowledge is required of the diffusion coefficients of the analytes in a diffusive layer. In this study, the diffusion coefficients of Hg(II), As(III), Mn(II), Mg(II), Cu(II), Cd(II) were determined in agarose gel and those of Ba(II), Cd(II), Cu(II), Mg(II), Mn(II) e Zn(II) in cellulose acetate membranes. These materials presented good performance and the reported results can be used as a data base for further DGT studies.

Determinação dos fatores neuroepidemiológicos das crises convulsivas em recém nascidos internados em unidades de terapia intensiva

O objetivo do nosso projeto foi determinar as características dos recém nascidos com crises convulsivas internados em unidades de terapia intensiva. Foi realizada uma pesquisa multicêntrica, observacional, prospectiva, cuja população alvo foi os recém nascidos com crises convulsivas internados em unidades de terapia intensiva, envolvendo equipe multidisciplinar constituída por neurologista infantil, neonatologistas, intensivistas pediátricos, enfermeiros, técnicos de enfermagem e fisioterapêutas. As crises foram definidas pelo critério clínico, com classificação de Volpe. Variáveis relacionadas à gestação, ao parto, características dos recém nascidos, aspectos das crises convulsivas e mortalidade foram analisadas. Estatística: descritiva (freqüências, medidas de tendência central e dispersão) e análise (teste de probabilidade, teste de risco e de acurácia). Comparamos as crises clínicas entre os recém nascidos de termo e pretermo e observamos diferenças com significância estatística na idade de início das crises, mais tardia nos prematuros, na etiologia predominante: hemorragia peri-intraventricular no prematuro e encefalopatia hipóxico-isquêmica no termo e tipo clínico de crise, clônica no pretermo e sutil no termo. Os testes de acurácia utilizados para determinar se o tipo clínico de crise convulsiva é predictor da etiologia da mesma não revelaram resultados positivos. Quanto às características associadas à mortalidade de prematuros com crise convulsiva, observamos associação entre ventilação mecânica e pneumonia com a mortalidade. Existem diferenças clínicas quando comparamos os recém nascidos pretermo e de termo com crises convulsivas, confirmando dados da literatura

Avaliação de um método de cromatografia em fase gasosa head space e estudo da estabilidade do etanol em amostras de sangue

The ethanol is the most overused psychoactive drug over the world; this fact makes it one of the main substances required in toxicological exams nowadays. The development of an analytical method, adaptation or implementation of a method known, involves a process of validation that estimates its efficiency in the laboratory routine and credibility of the method. The stability is defined as the ability of the sample of material to keep the initial value of a quantitative measure for a defined period within specific limits when stored under defined conditions. This study aimed to evaluate the method of Gas chromatography and study the stability of ethanol in blood samples, considering the variables time and temperature of storage, and the presence of preservative and, with that check if the conditions of conservation and storage used in this study maintain the quality of the sample and preserve the originally amount of analyte present. Blood samples were collected from 10 volunteers to evaluate the method and to study the stability of ethanol. For the evaluation of the method, part of the samples was added to known concentrations of ethanol. In the study of stability, the other side of the pool of blood was placed in two containers: one containing the preservative sodium fluoride 1% and the anticoagulant heparin and the other only heparin, was added ethanol at a concentration of 0.6 g/L, fractionated in two bottles, one being stored at 4ºC (refrigerator) and another at -20ºC (freezer), the tests were performed on the same day (time zero) and after 1, 3, 7, 14, 30 and 60 days of storage. The assessment found the difference in results during storage in relation to time zero. It used the technique of headspace associated with gas chromatography with the FID and capillary column with stationary phase of polyethylene. The best analysis of chromatographic conditions were: temperature of 50ºC (column), 150ºC (jet) and 250ºC (detector), with retention time for ethanol from 9.107 ± 0.026 and the tercbutanol (internal standard) of 8.170 ± 0.081 minutes, the ethanol being separated properly from acetaldehyde, acetone, methanol and 2-propanol, which are potential interfering in the determination of ethanol. The technique showed linearity in the concentration range of 0.01 and 3.2 g/L (0.8051 x + y = 0.6196; r2 = 0.999). The calibration curve showed the following equation of the line: y = x 0.7542 + 0.6545, with a linear correlation coefficient equal to 0.996. The average recovery was 100.2%, the coefficients of variation of accuracy and inter intra test showed values of up to 7.3%, the limit of detection and quantification was 0.01 g/L and showed coefficient of variation within the allowed. The analytical method evaluated in this study proved to be fast, efficient and practical, given the objective of this work satisfactorily. The study of stability has less than 20% difference in the response obtained under the conditions of storage and stipulated period, compared with the response obtained at time zero and at the significance level of 5%, no statistical difference in the concentration of ethanol was observed between analysis. The results reinforce the reliability of the method of gas chromatography and blood samples in search of ethanol, either in the toxicological, forensic, social or clinic

Alcoolemia em vítimas fatais de acidentes de trânsito no Rio Grande do Norte empregando cromatografia em fase gasosa - head space

Alcohol is one of the few psychotropic drugs that their consumption has admitted legally and sometimes encouraged by the society. Studies show alcohol as the highest consumption of drugs among young people and society in general, probably because of its availability and easy access. The abuse causes public health problems, which was closely related to the violence, socioeconomic problems and the high number of automobile accidents. Transit is one of the main sectors affected by the effects of alcohol, observing a high incidence in the studies. About half of automobile accidents occurs after the consumption of alcoholic beverage, and the vast majority of cases related to high concentrations of alcohol in the bloodstream. The relationship of drunk with traffic accidents is in fact evident everywhere in the world, including Brazil, where studies have shown a high relationship between alcohol consumption and traffic accidents. This study determined the alcohol in fatal victims of traffic accidents in the state of Rio Grande do Norte and established the profile of this population compared with those found in Brazil and other countries. Samples of blood of ethanol added to fulfillment of the standardization of chromatographic conditions and procedures for the analysis, being employed in the determination of alcohol in blood samples of 277 victims of traffic accidents, collected at the Institute of Scientific Technical Police of Rio Grande do North (ITEP) in the year 2007. The blood alcohol level was determined in these samples correlated with the sex, age and marital status of the victim and the location, day of week and month when the accident occurred, is doing a statistical analysis and outlining a profile of the victims of an accident at transit in the state of Rio Grande do Norte. The parameters of standardization studied ensured the quality of the analytical method and, consequently, to obtain reliable laboratory results. Being given the best temperature for injector (150 ºC), detector (250 ºC) and column (50 ºC) with a flow of gas in the column of 2mL/minutos and analysis of time of 12 minutes. The method was linear in the range of 0.01 to 3.2 g / L (r2 = 0.9989) with average recovery of 100.2% and precision with coefficient of variation less than 15%. The analysis carried out on victims of fatal road traffic accidents, ethanol detected in the blood in 66.43% of the victims and these, 96% showed concentration ≥ 0.2 g / L, 87.73% of victims were male, while 12.27% female. The younger age group (1535 years) was the most involved (52,35%) and most single (55.60%). The accidents occurred with greater prevalence in the day on Monday (27%) followed by Sunday (24,19%) and Saturday (15,52%) and it was found that the prevalence of injuries varied between the different months of the year, and in February (14.4%) and April (10.47%) the months that had a higher number of accidents, however this oscillation showed no statistically significant difference. Also no significant difference was observed between the tracks of concentration found in men and women. The standardized method showed to be efficient, given satisfactorily to the goals of this work, and the high levels of alcohol found in victims of fatal road traffic accidents are consistent with several studies of literature, and the profile of the victim also supported by presenting in its most young adults, male and single

Controle de qualidade físico-químico e biodisponibilidade relativa em ratos de suspensões orais de naproxeno sódico obtidas de farmácias de manipulação na cidade do Natal RN

Compounded medicines have been reported by the ANVISA due to decreased of the therapeutic response or toxicity of these formulations. The aim of this work was to investigate the physicochemical quality control among naproxen sodium oral suspensions 25 mg/mL obtained from six compounding pharmacies (A, B, C, D, E and F) and the manufactured suspension (R). In the quality control test, the tests of pH, content, homogeneity, volume and physical and organoleptic characteristics were performed according to the Brazilian Pharmacopoeia. The analytical method for determination of naproxen in suspensions was validate. This method showed excellent precision, accuracy, linearity and specificity. In the content test the suspensions B, C and E showed lower value and the F suspension showed a high value of the content. The products C and E were disapproved in the description of the physical and organoleptic characteristics test. In the pH test, three suspensions were outside specifications (C, E and F). Only the products R, A and D showed satisfactory results in these tests and therefore they were approved for relative bioavailability test. The R, A and D suspensions were orally administered to Wistar rats and the blood samples were taken at time intervals of 10, 20, 40, 60 min, 3, 4, 6, 24 and 48 h. The plasma samples were immediately stored at 80 ºC until analysis of HPLC. The bioanalytical method validation showed specificity, linearity (R2 0.9987), precision, accuracy, good recovery and stability. The chromatographic conditions were: flow rate of 1.2 mL.min-1 with a mobile phase of acetonitrile : sodium phosphate buffer pH 4.0 (50:50, v/v) at 280 nm, using a C18 column. The confidence interval of 90% for the Cmax and AUCt ratio was within the range of 80 - 125% proposed by the FDA. Only one suspension, obtained from the compounding pharmacy D, was considered bioequivalent to the rate of absorption under the conditions proposed by this study. Thus, the results indicate the need for strict supervision from the relevant authorities to ensure the patient safety and the quality of compounded drugs by pharmacies

Determinação dos períodos de convivência da cana-soca com plantas daninhas

Esta pesquisa foi realizada com o objetivo de determinar o período anterior à interferência (PAI) e o período total de prevenção à interferência (PTPI) das plantas daninhas em cana-de-açúcar (cana-soca, segundo corte) do cultivar SP803280. O experimento foi instalado sobre um Latossolo Vermelho. Os tratamentos experimentais constituíram de períodos crescentes de controle ou de convivência das plantas daninhas com a cultura: 0, 015, 0-34, 0-59. 0-90, 0-124, 0-157 e 0-269 dias (colheita da cana), totalizando 16 tratamentos, dispostos no delineamento de blocos ao acaso com quatro repetições. As plantas daninhas mais frequentes encontradas na área experimental foram: capim-colonião (Panicum maximum), carrapicho-de-carneiro (Acanthospermum hispidum) e apaga-fogo (Alternanthera tenella). Os resultados obtidos, admitindo-se 2 e 5% de redução, na produção de colmos da cana-de-açúcar revelaram que o período anterior à interferência (PAI) tolerado pela cultura foi de 9 e 18 dias após a brotação (DAB), e o período total de prevenção à interferência (PTPI), de 200 e 137 DAB, respectivamente, resultando num período crítico de prevenção à interferência de 9 aos 200 DAB para 2% e 18 aos 137 DAB para 5% de redução.

Eficiência do herbicida oxyfluorfen, quando veiculado ao papel, no controle de algumas espécies daninhas

Com o objetivo de encontrar um método alternativo de aplicação de herbicidas, estudou-se a eficiência do oxyfluorfen aplicado em três doses (0, 480 e 960g/ha) sob duas formas (pulverizado e veiculado em folha de papel) no controle em pré-emergência de Bidens pilosa, Desmodium tortuosum, Eleusine indica, Sida rhombifolia Amaranthus retroflexus, Acanthospermum hispidum e Digitaria horizontalis. A pulverização do oxyfluorfen foi realizada com o auxílio de um pulverizador, e sua veiculação foi feita através da determinação prévia da capacidade de embebição do papel. Obteve-se a concentração necessária da calda quando o papel atingiu completa embebição e então, escorrido o excesso, foi posto a secar na sombra. O delineamento experimental foi o inteiramente casualizado, com cinco repetições. Foram realizadas avaliações aos 30, 45 e 60 dias após a aplicação (DAA), e os resultados indicaram que os métodos de aplicação utilizados, assim como as doses, apresentaram excelente controle de B. pilosa, D. tortuosum, E. indica S. rhombifolia, D. horizontalis e A. retroflexus até 60 DAA, reduzindo tanto a densidade como o acúmulo de matéria seca. Quanto ao A. hispidum, ambos os métodos e doses não proporcionaram bom controle. Foi observado também que o papel utilizado como veículo do oxyfluorfen proporcionou redução na densidade de B. pilosa, E. indica, D. horizontalis e A. retroflexus e promoveu a germinação de S. rhombifolia e A. hispidum quando da avaliação aos DAA. Estes resultados demonstram a viabilidade de utilização de papel como veículo de aplicação do oxyfluorfen.

Determinação dos intervalos de referência para lipídios, lipoproteínas e apolipoproteínas no estado do Rio Grande do Norte

The lipid profile is a group of lab tests that include triglycerides, total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C). However, serum non-HDL-C, Apo A-I and Apo B levels, as well as the lipids ratios (TC/HDL-C, LDL-C/HDL-C and Apo B/Apo A-I), have been described as better predictors of cardiovascular diseases. Reference intervals are tools often used to help the evaluation of the people s health state. These days, Brazilian studies still use the reference intervals of lipids and lipoproteins from other countries, ignoring differences between the populations. Therefore, this study aimed to establish reference intervals for lipids, lipoproteins and apolipoproteins in adults of Rio Grande do Norte/Brazil. Healthy individuals (96 men and 283 women) between 18 and 59years old formed the reference sample group. The samples were collected after fasting 12 to 14 hours. Information on lifestyle and dietary habits of the participants were obtained through questionnaire. The serum glucose level and renal and liver activity were evaluated by laboratory testing. The results of lipid profile were analyzed according to sex, age and mesoregion of Rio Grande do Norte, with significance level of 5% (p < 0,05). The lower and upper reference limits were identified by the 2.5 percentile and 97.5 percentile, respectively, and assurance intervals of 90% was calculated for each of these limits. Among the determinants of lipid profile analyzed, only a few significant differences were observed according to sex, but in terms of age, the groups of smaller and older ages were most likely different. When evaluated by region, the means of West region shown the most significant variations. Not many studies were useful to compare the reference intervals determined in this study. Thus, it becomes necessary to carry out similar studies in other regions of Brazil and of the world given the clinical importance of reference intervals

Estudo de pré-formulação para dose fixa combinada dos tuberculostáticos rifampicina, isoniazida, pirazinamida e etambutol (4 em 1)

According to the global framework regarding new cases of tuberculosis, Brazil appears at the 18th place. Thus, the Ministry of Health has defined this disease as a priority in the governmental policies. As a consequence, studies concerning treatment and prevention have increased. Fixed-dose combination formulations (FDC) are recognized as beneficial and are recommended by WHO, but they present instability and loss on rifampicin bioavailability. The main purpose of this work was to carry out a pre-formulation study with the schedule 1 tuberculosis treatment drugs: rifampicin, isoniazid, pyrazinamide and ethambutol and pharmaceutical excipients (lactose, cellulose, magnesium stearate and talc), in order to develop an FDC product (150 mg of rifampicin + 75 mg of isoniazid + 400 mg of pyrazinamide + 250 mg of ethambutol). The studies consisted of the determination of particle size and distribution (Ferret s diameter) and shape through optical microscopy, as well as rheological and technological properties (bulk and tapped densities, Hausner Factor, Carr s Index, repose angle and flux rate) and interactions among drugs and drug excipient through thermal analysis (DSC, DTA, TG and your derivate). The results showed that, except isoniazid, the other drugs presented poor rheological properties, determined by the physical characteristics of the particles: small size and rod like particles shape for rifampicin; rectangular shape for pyrazinamide and ethambutol, beyond its low density. The 4 drug mixture also not presented flowability, particularly that one containing drug quantity indicated for the formulation of FDC products. In this mixture, isoniazid, that has the best flowability, was added in a lower concentration. The addition of microcrystalline cellulose, magnesium stearate and talc to the drug mixtures improved flowability properties. In DSC analysis probable interactions among drugs were found, supporting the hypothesis of ethambutol and pyrazinamide catalysis of the rifampicin-isoniazid reaction resulting in 3- formylrifamycin isonicotinyl hydrazone (HYD) as a degradation product. In the mixtures containing lactose Supertab® DSC curves evidenced incompatibility among drugs and excipient. In the DSC curves of mixtures containing cellulose MC101®, magnesium stearate and talc, no alterations were observed comparing to the drug profiles. The TG/DTG of the binary and ternary mixtures curves showed different thermogravimetrics profiles relating that observed to the drug isolated, with the thermal decomposition early supporting the evidences of incompatibilities showed in the DSC and DTA curves

Determinação de parâmetros de qualidade para formas farmacêuticas homeopáticas

All medicine, whether allopathic or homeopathic, must go through strict quality control, which must ratify their characteristics throughout the period of validity. During the time of preparation and storage, solutions of the drugs are in permanent contact with packaging materials that can release undesirable substances to the solution. Several factors may influence the release of packing materials, and factorial design (FD) is a useful tool for analyzing the phenomenon. The aim of this study was the determination of quality parameters for Homeopathic solid (globules) and liquid (drops) dosage forms. It was carried out analysis in homeopathic globules for weight variation, mechanical strength, and moisture content uniformity. For liquid preparations, standard solutions were prepared from natural rubber bulbs, which were subjected to exhaustive extraction with two ethanol solutions (30 and 70%) in the ultrasonic bath for 20 minutes at 25°C and 50°C in three successive cycles. Studies of transfer have been made within five days, by spectrophotometric analysis in the UV region at 312 nm with λmáx and 323 nm for samples in 70% ethanol and 30% respectively. PH values were analyzed. We also conducted two FD studies, where the first, the three-level variables were solvent (chloroform, ethanol and nhexane), sample mass (30, 60 and 90mg), particle size (large disk, small disk and powder sample). In the second study, the solvent level variables were different ethanolic degrees (EtOH 30%, 70% and pure). The percentage of lending in the solutions was 5.5%, 12.4%, 24.2% and 41% of the total estimated in the reference solution. The values of rate constants of transfer were determined in the order of 0.0134 days-1 and 0.0232 days-1 in absorbance values, the solutions in ethanol at 30% and 70% respectively. These results suggest that the speed of transfer of materials from rubber is affected both by the nature of the vehicle as by the temperature

Desenvolvimento e validação de método analítico para determinação de teor de ácido gálico e catequina no fitoterápico sanativo® por cromatografia líquida de alta eficiência

The herbal medicine Sanativo® is produced by the Pernambucano Laboratory since 1888 with indications of healing and hemostasis. It is composed of a fluid extract about Piptadenia colubrina, Schinus terebinthifolius, Cereus peruvianus and Physalis angulata. Among the plants in their composition, S. terebinthifolius and P. colubrina have in common phenolic compounds which are assigned most of its pharmacological effects. The tannins, gallic acid and catechin were selected as markers for quality control. The aim of this study was the development and validation of analytical method by HPLC/UV/DAD for the separation and simultaneous quantification of gallic acid (GAC) and catechin (CTQ) in Sanativo®. The chromatographic system was to stationary phase, C-18 RP column, 4,6 x 150 mm (5 mm) under a temperature of 35 ° C, detection at 270 and 210 nm. The mobile phase consisted of 0.05% trifluoroacetic acid and methanol in the proportions 88:12 (v/v), a flow rate of 1 ml/min. The analytical method presented a retention factor of 0.30 and 1.36, tail factor of 1.8 and 1.63 for gallic acid and catechin, respectively, resolution of 18.2, and theoretical plates above 2000. The method validation parameters met the requirements of Resolution n º 899 of May 29, 2003, ANVISA. The correlation coefficient of linear regression analysis for GAC and CTQ from the standard solution was 0.9958 and 0.9973 and when performed from the Sanativo® 0.9973 and 0.9936, the matrix does not interfere in the range 70 to 110 %. The limits of detection and quantification for GAC and CQT were 3.25 and 0.863, and 9.57 and 2.55 mg/mL, respectively. The markers, GAC and CQT, showed repetibility (coefficient of variation of 0.94 % and 2.36 %) and satisfactory recovery (100.02 ± 1.11 % and 101.32 ± 1.36 %). The method has been characterized selective and robust quantification of GAC and CTQ in the Sanativo® and was considered validated

Preparação e caracterização de sistemas emulsionados contendo benznidazol

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Avaliação da argila atapulgita para potencial uso como excipiente farmacêutico em formas sólidas

Clays are natural materials that have great potential for use as excipients for solid dosage forms. Palygorskite is a type of clay that has hydrophilic properties as well as a large surface area, which could contribute to the dissolution of drugs. Thus, the present study aims to evaluate the use of palygorskite clay, from Piaui (Northeast region of Brazil), as a pharmaceutical excipient for solid dosage forms, using rifampicin and isoniazid as the model drugs. The former is a poorly soluble drug often associated with isoniazid for tuberculosis treatment. Palygorskite was characterized by X-ray diffraction (XRD), X-ray fluorescence (XRF), particle size, transmission electron microscopy (TEM), scanning electron microscopy (SEM) and specific surface area (BET). The rheological and technological properties of palygorskite were determined and compared to those of talc, magnesium stearate and Aersosil 200. Mixtures between drugs and palygorskite were analyzed by differential scanning calorimetry (DSC), thermogravimetric analysis (TG) combined with thermal analysis (DTA) and Fourier Transform Infrared Spectroscopy (FT-IR), where the results were compared with those of the individual compounds. In addition, dissolution studies of solid dispersions and capsules containing the drugs, mixed with either palygorskite or a mixture of talc and magnesium stearate, were performed. The results showed that palygorskite has small particles with a high surface area. Its rheological characteristics were better than those of others commonly used glidants and lubricants. There was no interaction between palygorskite and the drugs (rifampicin and isoniazid). Among the dispersions studied, the mixture with palygorskite (5%) showed the highest drug dissolution when compared to other excipients. The dissolution of the rifampicin capsules containing palygosrkite was faster in higher concentrations. However, these differences were statistically different only in the first minutes of the dissolution experiment. The dissolution profile of isoniazid was also statistically different on the initial part of the experiment. The formulations prepared with isoniazid and palygorskite showed higher drug dissolution, but it was in descending order of concentration. According to these results, the palygorskite clay used in this study has great potential for application as an excipient for solid dosage forms

Desenvolvimento e validação de método analítico para determinação de WE010, WE014 e WE017

The synthetic guanylhydrazones WE010 (3,5-di-tert-butil-4-hidroxibenzaldehyde-guanylhydrazone), WE014 (4-bifenilcarboxialdehydeguanylhydrazone) and WE017 (3,4-diclorobenzaldehydeguanylhydrazone) showed high cytotoxic activity in terms of percentage inhibition of cancer cells growth. However, further progress in the development of these drug candidates requires precise and convenient methods for their qualitative and quantitative analyses. The aim of this study was to develop and validate High Performance Liquid Chromatography with diode-array detection (HPLC-DAD) and Ultra Fast Liquid Chromatography with diode-array detection (UFLC-DAD) methods suitable for as simultaneous as isolated determination of studied guanylhydrazones, based on the optimization of chromatographic parameters and obtaining reduced detection times. The chromatographic analyses of analytes by HPLC were performed on C18 ACE analytical column (150 mm x 4.6 mm), with a particle size of 5.0 μm. Among all the conditions assayed, the best results of separation were obtained with a mixture of methanol:water (60:40, v/v) as the mobile phase at a flow rate 1.5mL/min and pH of 3.5 adjusted at acetic acid. The UFLC method was developed by experimetal desing techniques in order to find optimal chromatographic analytical conditions, which were achieved on XR-ODS analytical column (50 mm x 3.0 mm), with a particle size of 2,2 μm, maintained at 25 ºC. The mobile phase was consisted of methanol:water (65:35, v/v) with 0.1% triethylamine (TEA) and pH of 3.5 adjusted at acetic acid, at a flow rate 0.5 mL/min. The procedure were validated following evaluating parameters such as specificity, linearity, limits of detection (LD) and quantification (LQ), precision, accuracy and robustness, giving results within the acceptable range. Although the UFLC method shows better sensitivity (lower values of LD and LQ), robustness (lower rates of relative standard deviation) and minimize spending time and solvent, both developed methods were adequately applied to the analysis of guanylhydrazones molecules, may be used in routine of quality control laboratories. Keywords: guanylhydrazones, HPLC/DAD, UFLC/DAD, validation of analitical method

DESENVOLVIMENTO DE CALOS em EXPLANTES DE CUPUAÇUZEIRO (Theobroma grandiflorum Schum.) em FUNÇÃO da CONCENTRAÇÃO DE AUXINAS E do MEIO LÍQUIDO

Objetivou-se estudar o efeito da concentração de auxina e do meio líquido sobre o desenvolvimento de calos de cupuaçuzeiro. Segmentos de eixos embrionários e cotilédones, obtidos de frutos de cupuaçu dos tipos Mamorana e Redondo, foram cultivados em 4 meios de cultura diferentes: 1) meio MS (50%), suplementado com 2,4-D (1; 2; 4; 8 mg/L); 2) sais N6 (SIGMA) (4 g/L), acrescidos de 2,4-D (0; 2; 4 mg/L) e ANA (0; 3; 5 mg/L); 3) igual ao anterior, suplementado apenas com ANA (3 mg/L); e 4) meio MS, acrescido com ANA (1 mM). Calos com aspecto branco e brilhante foram observados em segmentos de eixos embrionários e cotilédones, cultivados nas menores concentrações de meio 1 (1 e 2 mg/L), enquanto nas maiores concentrações (4 e 8 mg/L) se observou a formação de calos e massa calosa branco-opaca, em eixos embrionários e em segmentos cotiledonares, estas estruturas tornaram-se escuras dentro de oito semanas. Usando o meio 2, um grande número de raízes foram formadas, enquanto o mesmo meio suplementado apenas com ANA (3; 5 mg/L) originou uma massa calosa. A combinação de ANA e 2,4-D, 3 e 2 mg/L, respectivamente, promoveu a formação de calos brancos e raízes. A transferência das culturas para meio líquido, sem regulador de crescimento, promoveu aumento de tamanho dos explantes e escurecimento dos mesmos. O cultivo desses explantes no meio 3 resultou no aparecimento de calos amarelos, com aspecto friável, que permaneceram com a mesma aparência no meio 4.