Sensitive bromatometric assay methods for finasteride in pharmaceuticals

Three sensitive spectrophotometric methods are presented for the determination of finasteride in bulk and in tablets. The methods rely on the use of bromate-bromide reagent and three dyes namely, methyl orange, indigocarmine and thymol blue as reagents. They involve the addition of a measured excess of bromate-bromide reagent to finasteride in acid medium, and after the bromination reaction is judged to be complete, the unreacted bromine is determined by reacting with a fixed amount of either methylorange and measuring the absorbance at 520 nm (method A) or indigocarmine and measuring the absorbance at 610 nm (method B) or thymol blue and measuring the absorbance at 550 nm (method C). In all the methods, the amount of insitu generated bromine reacted corresponds to the amount of finasteride. The absorbance measured at the respective wavelength is found increase linearly with the concentration of finasteride. Beer's law is obeyed in the ranges 0.25- 2.0, 0.5-6.0 and 1-12 µg mL-1 for method A, method B and method C, respectively. The calculated molar absorptivity values are 5.7x10(4), 3.12x10(4) and 1.77x10(4) L mol-1 cm-1 respectively, for method A, method B and method C, and the corresponding Sandell sensitivity values are 0.0065, 0.012 and 0.021 µg cm-2. The limits of detection (LOD) and quantification (LOQ) are also reported for all the methods. Accuracy and, intra-day and inter-day precisions of the methods were established according to the current ICH guidelines. The methods were successfully applied to the determination of finasteride in commercially available tablets and the results were found to closely agree with the label claim. The results of the methods were statistically compared with those of a reference method by applying Student's t-test and F-test. The accuracy and reliability of the methods were further confirmed by performing recovery tests via standard addition procedure.

Ascorbic acid determination using a carbon paste electrode modified with iron(III) ions adsorbed on humic acid

An amperometric sensor was constructed, by using humic acids to immobilize Fe3+ ions on a carbon paste electrode (CPE-HA-Fe), and used for ascorbic acid (H2A) determination. The cyclic voltammogram of the electrode showed electrochemical response due to the Fe3+/Fe2+ couple at E1/2=+0.78 V vs SCE, using 0.5 mol L-1 KCl and 0.2 mol L-1 acetate/0.020 mol L-1 phosphate buffer, at pH = 5.4, as supporting electrolyte. When H2A is added to the electrolyte solution it is observed an oxidation process. The oxidation current, obtained by chronoamperommetry at +0.87 V vs SCE, is proportional to the concentration, represented by the equation I(µA) = 7.6286 [H2A] (mmol L-1) + 1.9583, r = 0.9996, for concentrations between 0.0 and 1.4 mmol L-1. The electrode showed high stability and was used for H2A determination in a natural orange juice.

Sensitive spectrophotometric assessment of carbofuran using dapsone as a new chromogenic reagent in formulations and environmental samples

A simple, sensitive and selective spectrophotometric method for the assessment of carbofuran in various formulations and in environmental water samples is described. The method is based on the coupling of hydrolyzed carbofuran with diazotized dapsone in alkaline medium at 0 ­ 4° C which gives orange red colored product having the absorption maximum at 480 nm. The product is stable for 48 h. Beer's law is obeyed in the concentration range of 0.1 ­ 4.0 µg ml-1. The molar absorptivity and Sandell's Sensitivity are 5.0 x 10(4) L mol-1 cm-1 and 4.4 ng cm-2 respectively. The method is highly reproducible and is confirmed by RSD values (1.144 %). From the recovery studies it is found that this method is accurate and it can be successfully employed for the determination of carbofuran.

Influência do método de inoculação, intensidade do ferimento e idade do fruto na severidade da podridão-de-cratera em melão

A podridão-de-cratera dos frutos de meloeiro, causada por Myrothecium roridum, vem ocorrendo com freqüência nos plantios da região Nordeste e ocasionando perdas de produção. Foi analisada a influência do método de inoculação (gota, pulverização, gota com ferimento, pulverização com ferimento e injeção subepidérmica), da intensidade (0, 1, 3, 5, 7, 9 e 10 ferimentos) e idade de ferimento (0, 3 e 6 horas) e da idade do fruto (12, 22 e 27 dias) na severidade da podridão-de-cratera em melão dos tipos Amarelo (cv. AF-682) e Honeydew (cv. Orange Flesh), inoculados com três isolados de M. roridum (CMM-609, CMM-636 e CMM-766). A severidade da doença foi influenciada pela interação entre métodos de inoculação, isolados e cultivares. As inoculações por pulverização ou deposição de gota propiciaram maiores lesões nos frutos submetidos a ferimentos. Entretanto, não foram observados sintomas nos frutos sem ferimentos. A inoculação por injeção subepidérmica, apesar de também provocar ferimento no fruto, apresentou lesões menores. A severidade da doença aumentou com o incremento do número de ferimentos, atingindo o máximo com 10 ferimentos. Verificou-se uma tendência de redução da severidade da doença nos frutos com o aumento da idade do ferimento. As lesões foram significativamente menores nos frutos feridos 6 horas antes da inoculação do que naqueles feridos imediatamente antes da inoculação. A idade do fruto não foi determinante para elevação ou redução da severidade da podridão-de-cratera.

Manejo da podridão de melão pelo controle do amadurecimento através do 1-mcp, sob duas condições de armazenamento

Objetivou-se neste trabalho avaliar o efeito de 1-MCP (300 nL.L¹) nas alterações fisiológicas que ocorrem durante o amadurecimento do melão, tipo Orange cv. Orange Flesh e sua influência no controle da podridão causada por Fusarium pallidoroseum, em dois ambientes de armazenamento, sem refrigeração (29 ± 1 ºC e umidade relativa de 65 ± 2 %) e refrigerado (10 ± 2 ºC e umidade relativa 90 ± 3 %) durante 15 dias e nove dias adicionais em condição ambiente. Avaliouse a atividade respiratória, produção de etileno, perda de matéria fresca, cor da casca e da polpa, firmeza da polpa, pH, acidez total titulável, sólidos solúveis totais, açúcares solúveis totais e severidade da doença. O delineamento foi inteiramente casualizado em arranjo fatorial com quatro repetições/tratamento. Frutos tratados com 1-MCP, armazenados em ambiente sem refrigeração, mantiveram firmeza da polpa praticamente inalterada até o 15º dia e quando em refrigeração mantiveram-se inalterados até o final da avaliação aos 24 dias após a colheita, retardando também a abscisão do pedúnculo, uma variável indicativa de maturação. O tratamento com 1-MCP retardou o amadurecimento de frutos controlando a podridão de F. pallidoroseum. Este tratamento também reduziu a respiração, produção, etileno, perda de peso, não influenciando significativamente as variáveis de qualidade do fruto: coloração da casca e polpa dos frutos, teor de sólidos solúveis totais e açúcares solúveis totais durante o período de armazenamento, bem como pH e acidez total titulável nas diferentes condições de armazenamento estudadas. A refrigeração interagiu positivamente com o 1-MCP, aumentando o tempo de conservação e sanidade do melão.

Chitosan and fungicides on postharvest control of Guignardia citricarpa and on quality of 'Pêra Rio' oranges

Citrus fruits are affected by the black spot disease caused by the fungus Guignardia citricarpa. Chitosan can be used as covering for fruits and may delay the ripening process and inhibit the growth of some fungi. Thus, the control of citrus black spot using chitosan and the fungicides thiabendazole and imazalil was assessed in addition to the physicochemical quality of 'Pêra Rio' oranges. The oranges were immersed into chitosan, thiabendazole or imazalil, and in chitosan mixed with both fungicides. The fruits were then stored at 25 °C, 80% RH, for 7 days and, after this storage period, subjected to physicochemical analyses. Chitosan in association with the fungicides reduced black spot in 'Pêra Rio' oranges and delayed the change in the orange skin colour from green to yellow during the postharvest storage. Total soluble solids, titratable acidity, pH, ascorbic acid content and ratio were not influenced by the treatments. Thus, chitosan applied with the fungicides thiabendazole and imazalil showed potential to control the development of black spot lesions on 'Pêra Rio' oranges during the postharvest period.

Terrain forms and spatial variability of soil properties in an area cultivatedwith citrus

The technique of precision agriculture and soil-landscape allows delimiting areas for localized management, allowing a localized application of agricultural inputs and thereby may contribute to preservation of natural resources. Therefore, the objective of this work was to characterize the spatial variability of chemical properties and clay content in the context of soil-landscape relationship in a Latosol (Oxisol) under cultivation of citrus. Soil samples were collected at a depth of 0.0-0.2 m in an area of 83.5 ha planted with citrus, as a 50-m intervals grid, with 129 points in concave terrain and 206 points in flat terrain, totaling 335 points. Values for the variables that express the chemical characteristics and clay content of soil properties were analyzed with descriptive statistics and geostatistical modeling of semivariograms for making maps of kriging. The values of range and kriging maps indicated higher variability in the shape of concave topography (top segment) compared with the shape of flat topography (slope and hillside segments below). The identification of different forms of terrain proved to be efficient in understanding the spatial variability of chemical properties and clay content of soil under cultivation of citrus.

Citrus root distribution under water stress grown in sandy soil of central Florida

The experiment was conducted in an orchard located in University of Florida (Citrus Research and Education Center), Lake Alfred, Polk County, Florida, USA. The objective of this study was to evaluate the effects of water stress in root distribution of 'Valencia' orange tree on 'Swingle' citrumelo rootstock. Three treatments were imposed on the trees: 1) normal irrigation with microsprinklers, 2) no irrigation in winter (November through mid-March) and 3) rainfall exclusion by placing a water repelling fabric (Tyvek) under the trees. Trees in treatments 1 and 2 received normal rainfall during the winter, but treatment 3 received no rain. Normal irrigation was resumed on all treatments in mid March. Soil was collected using root auger head (0.09 m diameter and height 0.25 m) in two opposing quadrants (West and East at 3 horizontal distances from tree trunk (1, 2 and 3 m) and 4 depths (0.0-0.15; 0.15-0.30; 0.30-0.60 and 0.60-0.90 m). The results from root sampling showed that there was a significant difference in root distribution between irrigated treatment and non irrigated/non rainfall.

Uso da eletroestimulação transcutânea para alívio da dor durante o trabalho de parto em uma maternidade-escola: ensaio clínico controlado

OBJETIVOS: determinar os desfechos maternos e neonatais de acordo com a aplicação ou não de estimulação elétrica transcutânea (EET) para alívio da dor do trabalho de parto antes da instalação da técnica combinada (raquianestesia + peridural). MÉTODOS: realizou-se ensaio clínico, randomizado, aberto, envolvendo 22 parturientes, com gestação a termo e feto único em apresentação cefálica, atendidas em hospital-escola de nível terciário em Recife, Brasil. Estas pacientes foram randomizadas para receber ou não EET antes da instalação da anestesia combinada (raquianestesia + anestesia peridural) para analgesia de parto. Avaliaram-se a intensidade da dor pela escala analógica visual (EAV), o tempo transcorrido entre a avaliação inicial e a necessidade de instalação da anestesia combinada, a duração do trabalho de parto, a freqüência de cesariana e parto instrumental, os escores de Apgar e a freqüência de hipóxia neonatal. Para análise estatística, foram utilizados os testes de Mann-Whitney e exato de Fisher, considerando-se o nível de significância de 5%. RESULTADOS: o tempo decorrido entre a avaliação da dor da parturiente e a necessidade de instalação da técnica combinada foi significativamente maior no grupo da EET (mediana de 90 minutos) quando comparado ao grupo controle (mediana de 30 minutos). A duração do trabalho de parto foi similar nos dois grupos (em torno de seis horas). Não houve diferença na evolução dos escores de EAV durante o trabalho de parto. A freqüência de cesariana foi de 18,2% nos dois grupos. Apenas um parto foi ultimado a fórcipe, no grupo controle. A mediana do escore de Apgar no quinto minuto foi 10, não se encontrando nenhum caso de hipóxia neonatal. CONCLUSÕES: a aplicação de EET foi efetiva em retardar a instalação da anestesia combinada para manter analgesia satisfatória durante o trabalho de parto, porém não apresentou efeito significativo sobre a intensidade da dor e a duração do trabalho de parto. Não houve efeitos deletérios maternos e neonatais.

Stop bugging me! : diversity in appearance of warning coloration

In nature, many animals use body coloration to communicate with each other. For example, colorations can be used as signals between individuals of the same species, but also to recognise individuals of other species, and if they may comprise a threat or not. Many animals use protective coloration to avoid predation. The two most common strategies of protective coloration are camouflage and aposematism. Camouflaged animals have coloration that minimises detection, usually by matching colours or structures in the background. Aposematic animals, on the other hand, signal to predators that they are defended. The defence can be physical structures, such as spikes and hairs, or chemical compounds that make the animal distasteful or even deadly toxic. In order for the warning signal to be effective, the predator has to recognise it as such. Studies have shown that birds for example, that are important visual predators on insects, learn to recognise and avoid unpalatable prey faster if they contrast the background or have large internal contrasts. Typical examples of aposematic species have conspicuous colours like yellow, orange or red, often in combination with black. My thesis focuses on the appearance and function of aposematic colour patterns. Even though researchers have studied aposematism for over a century, there is still a lot we do not know about the phenomenon. For example, as it is crucial that the predators recognise a warning signal, aposematic colorations should assumingly evolve homogeneously and be selected for maximal conspicuousness. Instead, there is an extensive variation of colours and patterns among warning colorations, and it is not uncommon to find typical cryptic colours, such as green and brown in aposematic colour patterns. One hypothesis to this variation is that an aposematic coloration does not have to be maximally signalling in order to be effective, instead it is sufficient to have distinct features that can be easily distinguished from edible prey. To be maximally conspicuous is one way to achieve this, but not the only way. Another hypothesis is that aposematic prey that do not exhibit maximal conspicuousness can exploit both camouflage and aposematism in a distance-dependent fashion, by being signalling when seen close up but camouflaged at a distance. Many prey animals also make use of both strategies by shifting colour at different ecological conditions such as seasonal variations, fluctuations in food resources or between life stages. Yet another explanation for the variation may be that prey animals are usually exposed to several predator species that vary in visual perception and tolerance towards various toxins. The aim with this thesis is, by studying their functions, to understand why aposematic warning signals vary in appearance, specifically in the level of conspicuousness, and if warning coloration can be combined with camouflage. In paper I, I investigated if the colour pattern of the aposematic larva of the Apollo butterfly (Parnassius apollo) can switch function with viewing distance, and be signalling at close range but camouflaged at a distance, by comparing detection time between different colour variants and distances. The results show that the natural coloration has a dual distance-dependent function. Moreover, the study shows that an aposematic coloration does not have to be selected for maximal conspicuousness. A prey animal can optimise its coloration primarily by avoiding detection, but also by investing in a secondary defence, which presence can be signalled if detected. In paper II, I studied how easily detected the coloration of the firebug (Pyrrhocoris apterus), a typical aposematic species, is at different distances against different natural backgrounds, by comparing detection time between different colour variants. Here, I found no distance-dependent switch in function. Instead, the results show that the coloration of the firebug is selected for maximal conspicuousness. One explanation for this is that the firebug is more mobile than the butterfly larva in study I, and movement is often incompatible with efficient camouflage. In paper III, I investigated if a seasonal related colour change in the chemically defended striated shieldbug (Graphosoma lineatum) is an adaptation to optimise a protective coloration by shifting from camouflage to aposematism between two seasons. The results confirm the hypothesis that the coloration expressed in the late summer has a camouflage function, blending in with the background. Further, I investigated if the internal pattern as such increased the effectiveness of the camouflage. Again, the results are in accordance with the hypothesis, as the patterned coloration was more difficult to detect than colorations lacking an internal pattern. This study shows how an aposematic species can optimise its defence by shifting from camouflage to aposematism, but in a different fashion than studied in paper I. The aim with study IV was to study the selection on aposematic signals by identifying characteristics that are common for colorations of aposematic species, and that distinguish them from colorations of other species. I compared contrast, pattern element size and colour proportion between a group of defended species and a group of undefended species. In contrast to my prediction, the results show no significant differences between the two groups in any of the analyses. One explanation for the non-significant results could be that there are no universal characteristics common for aposematic species. Instead, the selection pressures acting on defended species vary, and therefore affect their appearance differently. Another explanation is that all defended species may not have been selected for a conspicuous aposematic warning coloration. Taken together, my thesis shows that having a conspicuous warning coloration is not the only way to be aposematic. Also, aposematism and camouflage is not two mutually exclusive opposites, as there are prey species that exploit both strategies. It is also important to understand that prey animals are exposed to various selection pressures and trade-offs that affect their appearance, and determines what an optimal coloration is for each species or environment. In conclusion, I hold that the variation among warning colorations is larger and coloration properties that have been considered as archetypically aposematic may not be as widespread and representative as previously assumed.

Application of isotope-selective non-dispersive infrared spectrometry for the evaluation of the 13C-urea breath test: comparison with three concordant methods

The aim of this work was to compare the performance of isotope-selective non-dispersive infrared spectrometry (IRIS) for the 13C-urea breath test with the combination of the 14C-urea breath test (14C-UBT), urease test and histologic examination for the diagnosis of H. pylori (HP) infection. Fifty-three duodenal ulcer patients were studied. All patients were submitted to gastroscopy to detect HP by the urease test, histologic examination and 14C-UBT. To be included in the study the results of the 3 tests had to be concordant. Within one month after admission to the study the patients were submitted to IRIS with breath samples collected before and 30 min after the ingestion of 75 mg 13C-urea dissolved in 200 ml of orange juice. The samples were mailed and analyzed 11.5 (4-21) days after collection. Data were analyzed statistically by the chi-square and Mann-Whitney test and by the Spearman correlation coefficient. Twenty-six patients were HP positive and 27 negative. There was 100% agreement between the IRIS results and the HP status determined by the other three methods. Using a cutoff value of delta-over-baseline (DOB) above 4.0 the IRIS showed a mean value of 19.38 (minimum = 4.2, maximum = 41.3, SD = 10.9) for HP-positive patients and a mean value of 0.88 (minimum = 0.10, maximum = 2.5, SD = 0.71) for negative patients. Using a cutoff value corresponding to 0.800% CO2/weight (kg), the 14C-UBT showed a mean value of 2.78 (minimum = 0.89, maximum = 5.22, SD = 1.18) in HP-positive patients. HP-negative patients showed a mean value of 0.37 (minimum = 0.13, maximum = 0.77, SD = 0.17). IRIS is a low-cost, easy to manage, highly sensitive and specific test for H. pylori detection. Storing and mailing the samples did not interfere with the performance of the test.

α-Pinene oxide and verbenol oxide isomerizations over heterogeneous catalysts

Terpenes are a valuable natural resource for the production of fine chemicals. Turpentine, obtained from biomass and also as a side product of softwood industry, is rich in monoterpenes such as α-pinene and β-pinene, which are widely used as raw materials in the synthesis of flavors, fragrances and pharmaceutical compounds. The rearrangement of their epoxides has been thoroughly studied in recent years, as a method to obtain compounds which are further used in the fine chemical industry. The industrially most desired products of α-pinene oxide isomerization are campholenic aldehyde and trans-carveol. Campholenic aldehyde is an intermediate for the manufacture of sandalwood-like fragrances such as santalol. Trans-carveol is an expensive constituent of the Valencia orange essence oil used in perfume bases and food flavor composition. Furthermore it has been found to exhibit chemoprevention of mammary carcinogenesis. A wide range of iron and ceria supported catalysts were prepared, characterized and tested for α-pinene oxide isomerization in order to selective synthesis of above mentioned products. The highest catalytic activity in the preparation of campholenic aldehyde over iron modified catalysts using toluene as a solvent at 70 °C (total conversion of α-pinene oxide with a selectivity of 66 % to the desired aldehyde) was achieved in the presence of Fe-MCM-41. Furthermore, Fe-MCM-41 catalyst was successfully regenerated without deterioration of catalytic activity and selectivity. The most active catalysts in the synthesis of trans-carveol from α-pinene oxide over iron and ceria modified catalysts in N,N-dimethylacetamide as a solvent at 140 °C (total conversion of α-pinene oxide with selectivity 43 % to trans-carveol) were Fe-Beta-300 and Ce-Si-MCM-41. These catalysts were further tested for an analogous reaction, namely verbenol oxide isomerization. Verbenone is another natural organic compound which can be found in a variety of plants or synthesized by allylic oxidation of α-pinene. An interesting product which is synthesized from verbenone is (1R,2R,6S)-3-methyl-6-(prop-1-en-2-yl)cyclohex-3-ene-1,2-diol. It has been discovered that this diol possesses potent anti-Parkinson activity. The most effective way leading to desired diol starts from verbenone and includes three stages: epoxidation of verbenone to verbenone oxide, reduction of verbenone oxide and subsequent isomerization of obtained verbenol oxide, which is analogous to isomerization of α-pinene oxide. In the research focused on the last step of these synthesis, high selectivity (82 %) to desired diol was achieved in the isomerization of verbenol oxide at a conversion level of 96 % in N,N-dimethylacetamide at 140 °C using iron modified zeolite, Fe-Beta-300. This reaction displayed surprisingly high selectivity, which has not been achieved yet. The possibility of the reuse of heterogeneous catalysts without activity loss was demonstrated.

Nephrotoxicity of Bence-Jones proteins: interference in renal epithelial cell acidification

The aim of the present study was to evaluate the acidification of the endosome-lysosome system of renal epithelial cells after endocytosis of two human immunoglobulin lambda light chains (Bence-Jones proteins, BJP) obtained from patients with multiple myeloma. Renal epithelial cell handling of two BJP (neutral and acidic BJP) was evaluated by rhodamine fluorescence. Renal cells (MDCK) were maintained in culture and, when confluent, were incubated with rhodamine-labeled BJP for different periods of time. Photos were obtained with a fluorescence microscope (Axiolab-Zeiss). Labeling density was determined on slides with a densitometer (Shimadzu Dual-Wavelength Flying-Spot Scanner CS9000). Endocytosis of neutral and acidic BJP was correlated with acidic intracellular compartment distribution using acridine orange labeling. We compared the pattern of distribution after incubation of native neutral and acidic BJP and after complete deglycosylation of BJP by periodate oxidation. The subsequent alteration of pI converted neutral BJP to acidic BJP. There was a significant accumulation of neutral BJP in endocytic structures, reduced lysosomal acidification, and a diffuse pattern of acidification. This pattern was reversed after total deglycosylation and subsequent alteration of the pI to an acidic BJP. We conclude that the physicochemical characteristics of BJP interfere with intracellular acidification, possibly explaining the strong nephrotoxicity of neutral BJP. Lysosomal acidification is fundamental for adequate protein processing and catabolism.

Effect of actinomycin D on simian rotavirus (SA11) replication in cell culture

Rotaviruses are the major cause of viral diarrhea in humans and animals. Actinomycin D (Act D) is an antibiotic that intercalates DNA and therefore inhibits DNA-dependent transcription. The current study was carried out to assess the influence of Act D on the replication of simian rotavirus (SA11) in cell culture. Virus-infected MA-104 cell cultures were studied in the presence of Act D at concentrations of 1.25 and 2.5 µg/ml. Treatment of rotavirus-infected cells with 2.5 µg/ml Act D 48 h post-infection reduced the cytoplasmic metachromasia after staining with acridine orange by 25%. Viral RNA labeled with ³H-uridine in the presence of the drug was separated by polyacrylamide gel electrophoresis. Viral RNA replication was not affected by Act D, but increased ³H-uridine uptake was demonstrable by infected cells in the presence of the drug. This possibly was due to the inhibition of cellular RNA synthesis by Act D, which thus enhances incorporation of the radionuclide into the viral RNA. Act D reduced the number of infected cells presenting virus-specific fluorescence 48 h post-infection by more than 50%. These data suggest that Act D may have complexed with viral RNA and prevented newly synthesized mRNA from being translated, but may not have prevented early replication.

Induction of liver monooxygenases by annatto and bixin in female rats

Annatto or urucum is an orange-yellow dye obtained from Bixa orellana seeds. It has been used as a natural dye in a variety of food products, drugs and cosmetics, and also in Brazilian cuisine as a condiment ('colorau'). Bixin, a carotenoid devoid of provitamin A activity, is the main pigment found in annatto. Some carotenoids (canthaxanthin, astaxanthin and ß-Apo-8'-carotenal) are known to be potent inducers of CYP1A1, a property not shared by others (ß-carotene, lycopene and lutein). Little is known, however, about the CYP1A1-inducing properties of bixin and annatto. The present study was performed to determine the effects of an annatto extract (28% bixin) and bixin (95% pure) on rat liver monooxygenases. Adult female Wistar rats were treated by gavage with daily doses of annatto (250 mg/kg body weight, which contains approximately 70 mg bixin/kg body weight), bixin (250 mg/kg body weight) or the vehicle only (corn oil, 3.75 g/kg body weight) for 5 consecutive days, or were not treated (untreated control). The activities of aniline-4-hydroxylase (A4H), ethoxycoumarin-O-deethylase (ECOD), ethoxy- (EROD), methoxy- (MROD), pentoxy- (PROD) and benzyloxy- (BROD) resorufin-O-dealkylases were measured in liver microsomes. Annatto (250 mg/kg containing 70 mg bixin/kg) induced EROD (3.8x), MROD (4.2x), BROD (3.3x) and PROD (2.4x). Bixin (250 mg/kg) was a weaker inducer of EROD (2.7x), MROD (2.3x) and BROD (1.9x) and did not alter PROD, A4H or ECOD activities. These results suggest that constituents of the extract other than bixin play an important role in the induction of CYP1A and CYP2B observed with annatto food colorings.