Avaliação da concentração de retinol em fígados frescos e congelados de frangos das linhagens Cobb e Ross

The intake of adequate quantities of food, including those rich in vitamins, is necessary for a healthy life. The lack of vitamin A has been characterized as a public health problem in developing countries, however, a high intake of vitamin A can result in toxic and teratogenics effects. High concentrations of vitamin A have been observed in the livers of animals. The objective of this study was to assess the levels of retinol in chicken livers and verify the effect of frozen storage on these levels. 64 livers from two chicken strains, Cobb and Ross, were used, came from four different farms. We examined 32 livers from each strain, 8 samples from each farm. Liver sample were homogenized individually, then 4 aliquots were taken from each sample. One of aliquots was analyzed immediately after slaughter (T0), the others were analyzed after 30, 60 and 90 days of storage at -18oC (T30, T60 and T90, respectively). Retinol dosage in the liver was determined by High Performance Liquid Chromatography (HPLC). The levels of retinol varied significantly according to the strain. The mean retinol value in the fresh samples was 6678.0 ± 1337.7 and 8324.1 ± 1158.5 µg/100g in the Cobb and Ross strain, respectively. Values of 4258 ± 918.7 ± 1391.7 and 4650.5 ± 1391.7 μg/100g were found after 90 days of storage for Cobb and Ross strain, respectively. The liver freezing caused a significant reduction in their levels of retinol, causing a loss of up to 44% with respect to fresh livers. The reduction in retinol levels occurred from 30 days of storage. Even with the losses from the frozen, the ingestion of a typical portion of 100 g of liver, regardless the chicken strain analyzed, surpass all recommendations of consumption and the maximum tolerable intake of vitamin A (3000 μg/day) for adults

Avaliação de megadoses de retinol palmitato no pós-parto imediato sobre o retinol do leite de puérperas atendidas no hospital josé pedro bezerra, Natal-RN

Vitamin A deficiency (VAD) is a serious public health problem in developing countries, and as a therapeutic and prophylactic measure retinil palmitate is being supplemented. Nevertheless its efficacy has been questioned. The objective of the study was to evaluate the supplementation of two retinil palmitate megadosis on the serum retinol levels of post partum healthy mothers from Dr. José Pedro Bezerra (Hospital Santa Catarina) hospital, Natal - RN. The enrolled women (n=199) were randomly distributed into three studied groups and supplemented with retinil palmitate immediately after delivery with a single 200,000 IU dose (group S1), two 200,000 IU dose (group S2) with 24h difference between the doses, or no supplementation (group C). Among women selected, 143 remained until the end of the study. The influence of vitamin A dietary intake was evaluated during pregnancy and after 30 days of delivery. The average intake of the population was reasonable, but a high prevalence of inadequate intake was found. Retinol in colostrums and mature milk was determined by high performance liquid chromatography (HPLC). The retinol average in colostrums and mature milk in the supplemented and control groups were adequate according to the reference values. In colostrums, women from groups C, S1 and S2 presented retinol averages by milk volume of 94.8 ± 40.2 µg/dL, 92.2 ± 50.0 µg/dL and 91.8 ± 53.7 µg/dL, respectively. No difference was found between these averages (p=0.965), this was also seen when the values where expressed as µg/g of fat (p=0.905). After 30 days of delivery, retinol per milk volume differed between the control group (36.6 ± 17.5 µg/dL) and groups supplemented with 200,000 IU (51.0 ± 28.8 µg/dL) or 400,000 IU (55.2 ± 31.6 µg/dL) of retinil palmitate (p<0,05). Nevertheless, when S1 and S2 groups where compared, no significant difference was found (p=0.97). Considering retinol/g of fat, the means were 12.7 ± 6.7 µg/g, 15.6 ± 8.3 µg/g and 17.2 ± 8.9 µg/g for groups C, S1 and S2, respectively, with significant difference between groups S2 and C (p=0,01). Subclinical VAD prevalence showed a serious public health problem in the study population (32% in colostrums and 31.5% in mature milk). When analyzing the groups separately, the group which received two doses (200,000 IU + 200,000 IU) presented the lowest VAD prevalence (20.7%). Retinil palmitate supplementations of 200,000 IU and 400,000 IU (divided in two doses) in the immediate post partum showed no significant difference. Nevertheless, the 400,000 IU (divided in two doses) supplementation showed a reduction in VAD

Avaliação da suplementação com vitamina E, na forma natural ou sintética, em mulheres no pós-parto imediato e sua concentração no colostro

The Vitamin E consists of eight chemically homologous forms, designated alpha, beta, gamma and delta tocopherols and tocotrienols. Biologically, the alpha-tocopherol (α-TOH) is the most important. Commercially, are found two types of α-TOH a natural (RRR-alpha-tocopherol) and another synthetic (all-rac-alpha-tocopherol). Both forms are absorbed in the intestine, the liver is a preference in favor of forms 2R, due to transfer protein α-TOH. It has higher affinity to these stereoisomers. Newborns are considered high risk for vitamin E deficiency, mainly premature, these have breast milk as a food source for maintenance of serum α-TOH. Clinical signs such as thrombocytosis, hemolytic anemia, retrolental fibroplasia, intraventricular hemorrhage, bronchopulmonary dysplasia and spinocerebellar degeneration can be found in case of a low intake of α-TOH. Thus, maternal supplementation on postpartum with α-TOH can be an efficient way to increase levels of vitamin E in breast milk and thus the consequently increase the supply of micronutrient for the newborn. However, most studies with vitamin E supplementation have been conducted in animals and little is known about the effect of maternal supplementation in humans, as well as on its efficiency to increase levels of α-TOH in human milk, depending on the shape natural or synthetic. The study included 109 women, divided into three groups: control without supplementation (GC) (n=36), supplemented with natural capsule (GNAT) (n=40) and the synthetic capsule (GSINT) (n=33). Blood samples were collected for determination of maternal nutritional status, and colostrums at initial contact and after 24 hours post-supplementation. Analyses were performed by High Performance Liquid Chromatography. Values of α-TOH in serum below 499.6mg/dL were considered deficient. We used the Kruskal-Wallis test and Tukey test to confirm the increase of alpha-tocopherol in milk and efficiency of administered capsules. Daily consumption of α-TOH was based on daily intake of 500 mL of colostrum by the newborn and compared with the nutritional requirement for children from 0 to 6 months of age, 4 mg / day. The mothers had mean concentration of serum α-TOH in 1016 ± 52, 1236 ± 51 and 1083 ± 61 mg / dL, in CG, GNAT and GSINT respectively. There were no women with deficiiency. The GC did not change the concentrations of α-TOH in colostrum. While women supplemented with natural and synthetic forms increased concentrations of α-TOH colostrum in 57.6% and 39%, respectively. By comparing supplemented groups, it was observed a significant difference (p=0.04), the natural capsule more efficient than the synthetic, approximately 49.6%. Individually, 21.1% of the women provided below 4mg/day of α-TOH, after supplementation for this index declined4.1%. Thus, maternal supplementation postpartum raised the levels of alpha-tocopherol in colostrum, and increased efficiency was observed with the natural form

Caracterização e quantificação de marcadores químicos do extrato hidroetanólico das folhas de Kalanchoe brasiliensis Cambess

Kalanchoe brasiliensis Cambess (Crassulaceae), commonly known as saião , coirama branca , folha grossa , is originally from Brazil and commonly found in São Paulo to Bahia, mainly in the coastal zone. Regarding of biological activities, most preclinical studies were found in the literature, mainly about the anti-inflammatory activity of extracts obtained from leaves and / or aerial parts of K. brasiliensis. As regards the chemical constitution, it has been reported mainly the presence of flavonoids in the leaves of the species, but until this moment did not knows which are the active compounds. Although it is a species widely used in traditional medicine in Brazil, there is no monograph about the quality parameters of the plant drug. In this context, this study aims to characterize and quantify the chemical markers of hydroethanolic extract (HE) from the leaves of K. brasiliensis, which can be used in quality control of plant drug and derivatives obtained from this species. The methodology was divided into two parts: i. Phytochemical study: to fractionate, isolate and characterizate of the chemical (s) marker (s) of the HE from the leaves of K. brasiliensis; ii. To Developed validate of analytical method by High Performance Liquid Chromatography (HPLC)-diode array detector (DAD) to quantify the chemical (s) marker (s) of the EH. i. The EH 50% was prepared by turbo extraction method. It was then submitted to liquid-liquid partition, obtaining dichloromethane, n-butanol and ethyl acetate (AcOEt) fractions. The AcOEt fraction was selected to continue the fractionation process, because it has a chemical profile rich in flavonoids. The acOEt fraction was submitted to column chromatography using different systems for obtaining the compound Kb1. To identify this compound, it was submitted to UV analysis ii. For quantitative analysis, the EH was analyzed by HPLC, using different methods. After selecting the most appropriate method, which showed satisfactory resolution and symmetrical peaks, it was validated according to parameters in the RE 899/2003. As result, it was obtained from the AcOEt fraction the compound Kb1 (2.7 mg). Until this moment, the basic nucleus was characterized by UV analysis using shift reagents. The partial chemical structure of the compound Kb1 was identified as a flavonol, containing hydroxyls in 3 , 4 position (ring A), 5 and 7 free (ring B) and a replacement of the C3 hydroxyl by a sugar. As the analysis were performed in the HPLC coupled to a DAD, we observed that the UV spectrum of the major peaks of EH from K. brasiliensis shown similar UV spectrum. According to the literature, it has been reported the presence of patuletin glycosydes derivatives in the leaves of this species. Therefore, it is suggested that the compound Kb1 is glycosylated patuletin derivative. Probably the sugar (s) unit(s) are linked in the C3 in the C ring. . Regarding the development of HPLC analytical method, the system used consists of phase A: water: formic acid (99,7:0,3, v / v) and phase B: methanol: formic acid (99,7:0,3, v / v), elution gradient of 40% B - 58% B in 50 minutes, ccolumn (Hichrom ®) C18 (250x4, 0 mm, 5 μm), flow rate 0.8 mL / min, UV detection at 370 nm, temperature 25 ° C. In the analysis performed with the co-injection of thecompound Kb1 + HE of K. brasiliensis was observed that it is one of the major compounds with a retention time of 12.47 minutes and had a content of 15.3% in EH of leaves from K. brasiliensis. The method proved to be linear, precise, accurate and reproducible. According to these results, it was observed that compound Kb1 can be used as a chemical marker of EH from leaves of K. brasiliensis, to assist in quality control of drug plant and its derivatives

Avaliação da concentração de retinol em fígados frescos e congelados de frangos das linhagens Cobb e Ross

The intake of adequate quantities of food, including those rich in vitamins, is necessary for a healthy life. The lack of vitamin A has been characterized as a public health problem in developing countries, however, a high intake of vitamin A can result in toxic and teratogenics effects. High concentrations of vitamin A have been observed in the livers of animals. The objective of this study was to assess the levels of retinol in chicken livers and verify the effect of frozen storage on these levels. 64 livers from two chicken strains, Cobb and Ross, were used, came from four different farms. We examined 32 livers from each strain, 8 samples from each farm. Liver sample were homogenized individually, then 4 aliquots were taken from each sample. One of aliquots was analyzed immediately after slaughter (T0), the others were analyzed after 30, 60 and 90 days of storage at -18oC (T30, T60 and T90, respectively). Retinol dosage in the liver was determined by High Performance Liquid Chromatography (HPLC). The levels of retinol varied significantly according to the strain. The mean retinol value in the fresh samples was 6678.0 ± 1337.7 and 8324.1 ± 1158.5 µg/100g in the Cobb and Ross strain, respectively. Values of 4258 ± 918.7 ± 1391.7 and 4650.5 ± 1391.7 μg/100g were found after 90 days of storage for Cobb and Ross strain, respectively. The liver freezing caused a significant reduction in their levels of retinol, causing a loss of up to 44% with respect to fresh livers. The reduction in retinol levels occurred from 30 days of storage. Even with the losses from the frozen, the ingestion of a typical portion of 100 g of liver, regardless the chicken strain analyzed, surpass all recommendations of consumption and the maximum tolerable intake of vitamin A (3000 μg/day) for adults

Avaliação de megadoses de retinol palmitato no pós-parto imediato sobre o retinol do leite de puérperas atendidas no Hospital José Pedro Bezerra, Natal-RN

Vitamin A deficiency (VAD) is a serious public health problem in developing countries, and as a therapeutic and prophylactic measure retinil palmitate is being supplemented. Nevertheless its efficacy has been questioned. The objective of the study was to evaluate the supplementation of two retinil palmitate megadosis on the serum retinol levels of post partum healthy mothers from Dr. José Pedro Bezerra (Hospital Santa Catarina) hospital, Natal - RN. The enrolled women (n=199) were randomly distributed into three studied groups and supplemented with retinil palmitate immediately after delivery with a single 200,000 IU dose (group S1), two 200,000 IU dose (group S2) with 24h difference between the doses, or no supplementation (group C). Among women selected, 143 remained until the end of the study. The influence of vitamin A dietary intake was evaluated during pregnancy and after 30 days of delivery. The average intake of the population was reasonable, but a high prevalence of inadequate intake was found. Retinol in colostrums and mature milk was determined by high performance liquid chromatography (HPLC). The retinol average in colostrums and mature milk in the supplemented and control groups were adequate according to the reference values. In colostrums, women from groups C, S1 and S2 presented retinol averages by milk volume of 94.8 ± 40.2 µg/dL, 92.2 ± 50.0 µg/dL and 91.8 ± 53.7 µg/dL, respectively. No difference was found between these averages (p=0.965), this was also seen when the values where expressed as µg/g of fat (p=0.905). After 30 days of delivery, retinol per milk volume differed between the control group (36.6 ± 17.5 µg/dL) and groups supplemented with 200,000 IU (51.0 ± 28.8 µg/dL) or 400,000 IU (55.2 ± 31.6 µg/dL) of retinil palmitate (p<0,05). Nevertheless, when S1 and S2 groups where compared, no significant difference was found (p=0.97). Considering retinol/g of fat, the means were 12.7 ± 6.7 µg/g, 15.6 ± 8.3 µg/g and 17.2 ± 8.9 µg/g for groups C, S1 and S2, respectively, with significant difference between groups S2 and C (p=0,01). Subclinical VAD prevalence showed a serious public health problem in the study population (32% in colostrums and 31.5% in mature milk). When analyzing the groups separately, the group which received two doses (200,000 IU + 200,000 IU) presented the lowest VAD prevalence (20.7%). Retinil palmitate supplementations of 200,000 IU and 400,000 IU (divided in two doses) in the immediate post partum showed no significant difference. Nevertheless, the 400,000 IU (divided in two doses) supplementation showed a reduction in VAD

Avaliação da suplementação com vitamina E, na forma natural ou sintética, em mulheres no pós-parto imediato e sua concentração no colostro

The Vitamin E consists of eight chemically homologous forms, designated alpha, beta, gamma and delta tocopherols and tocotrienols. Biologically, the alpha-tocopherol (α-TOH) is the most important. Commercially, are found two types of α-TOH a natural (RRR-alpha-tocopherol) and another synthetic (all-rac-alpha-tocopherol). Both forms are absorbed in the intestine, the liver is a preference in favor of forms 2R, due to transfer protein α-TOH. It has higher affinity to these stereoisomers. Newborns are considered high risk for vitamin E deficiency, mainly premature, these have breast milk as a food source for maintenance of serum α-TOH. Clinical signs such as thrombocytosis, hemolytic anemia, retrolental fibroplasia, intraventricular hemorrhage, bronchopulmonary dysplasia and spinocerebellar degeneration can be found in case of a low intake of α-TOH. Thus, maternal supplementation on postpartum with α-TOH can be an efficient way to increase levels of vitamin E in breast milk and thus the consequently increase the supply of micronutrient for the newborn. However, most studies with vitamin E supplementation have been conducted in animals and little is known about the effect of maternal supplementation in humans, as well as on its efficiency to increase levels of α-TOH in human milk, depending on the shape natural or synthetic. The study included 109 women, divided into three groups: control without supplementation (GC) (n=36), supplemented with natural capsule (GNAT) (n=40) and the synthetic capsule (GSINT) (n=33). Blood samples were collected for determination of maternal nutritional status, and colostrums at initial contact and after 24 hours post-supplementation. Analyses were performed by High Performance Liquid Chromatography. Values of α-TOH in serum below 499.6mg/dL were considered deficient. We used the Kruskal-Wallis test and Tukey test to confirm the increase of alpha-tocopherol in milk and efficiency of administered capsules. Daily consumption of α-TOH was based on daily intake of 500 mL of colostrum by the newborn and compared with the nutritional requirement for children from 0 to 6 months of age, 4 mg / day. The mothers had mean concentration of serum α-TOH in 1016 ± 52, 1236 ± 51 and 1083 ± 61 mg / dL, in CG, GNAT and GSINT respectively. There were no women with deficiiency. The GC did not change the concentrations of α-TOH in colostrum. While women supplemented with natural and synthetic forms increased concentrations of α-TOH colostrum in 57.6% and 39%, respectively. By comparing supplemented groups, it was observed a significant difference (p=0.04), the natural capsule more efficient than the synthetic, approximately 49.6%. Individually, 21.1% of the women provided below 4mg/day of α-TOH, after supplementation for this index declined4.1%. Thus, maternal supplementation postpartum raised the levels of alpha-tocopherol in colostrum, and increased efficiency was observed with the natural form

Efeito Da Suplementação Materna Com Palmitato De Retinila Sobre A Concentração De Retinol No Colostro Em Condições De Jejum E Pós-Prandial.

Vitamin A is important in many essential body processes and its deficiency results in serious consequences for human health. Breast milk is the only source of this vitamin for children that are exclusively breastfed. Analysis of vitamin A in mother s milk is important because its concentration is related to maternal vitamin A status and to its ingestion by the mother during pregnancy. The aim of the present study was to assess the effect of maternal supplementation with retynil palmitate on the concentration of colostrum retinol under fasting and postprandial conditions. A total of 149 nursing mothers were recruited at the Januário Cicco Maternity School (Natal, Brazil) and allocated to two groups: Comparison (n = 69) and Test (n = 80). Blood and colostrum (in fasting and postprandial conditions) samples were collected up to 24hs after delivery. Serum retinol and colostrum levels were analyzed by high-performance liquid chromatography. The serum retinol level of 41.6 ± 12.7μg/dL (mean ± standard deviation) indicates adequate biochemical nutritional status. Colostrum retinol level was not influenced by serum retinol levels under any of the conditions established. In the colostrum, the retinol concentration in the unsupplemented test group was 67.3 ± 37.7 μg/dL under fasting and 80.3 ± 35.1 μg/dL under postprandial conditions (p<0.05), showing an increase of 19.3%. In the supplemented test group the values were 102.6 ± 57.3 μg/dL and 133.4 ± 78.3 μg/dL under fasting and postprandial, respectively (p<0.05), representing an increase of 30%. Considering that under fasting conditions most of the vitamin A transported to the milk originates in the retinol binding protein (RBP), the postprandial increase in colostrum retinol suggests a different transport mechanism of retinol to maternal milk from that performed by RBP. This situation becomes more evident under supplementation conditions.

Efeito do estado nutricional bioquímico de retinol e alfatocoferol sobre seus níveis em lactantes

Vitamins A and E are essential nutrients in many biological processes, so that their adequate supply to the neonate is crucial. However, the bioavailability of vitamins may be limited by factors such as maternal nutritional status and the interaction between nutrients. This study aimed to investigate the effect of biochemical nutritional status of retinol and alpha-tocopherol levels in serum and colostrum. The study included 103 healthy puerperal women treated at the reference state maternity hospital (Natal-RN). Colostrum and serum samples were collected fasting in the immediate postpartum period and the analysis of retinol and alpha-tocopherol were determined by high-performance liquid chromatography. Specific cutoff points were adopted to characterize the biochemical status of vitamins A and E. For the total group of lactanting women the average concentration of retinol in serum (1.49 ± 0.4 μmol/L-1) and colostrum (2.18 ± 0.8 μmol/L-1), as well as alpha-tocopherol in serum (26.4 ± 8.0 μmol/L-1) and colostrum (26.1 ± 12.8 μmol/L-1), indicated adequate biochemical state. However, when evaluating the individual, was found a high prevalence of deficient serum (15%) and colostrum retinol (50%), and also alphatocopherol in serum (16%) and colostrum (61%). In women with serum retinol ≥ 1.05 μmol/L-1, found an inverse correlation between serum retinol and alpha-tocopherol in colostrum (p = 0.008, r = -0.28). This association was not observed in women with serum retinol <1.05 μmol/L-1. This situation demonstrates for the first time in humans that high physiological levels of serum retinol, without supplementation, can negatively influence the transfer of alpha-tocopherol in breast milk. Although the diagnosis of satisfactory nutritional status lactanting women showed high risk of subclinical deficiency of vitamins A and E from measurements made in the colostrum

Influência da suplementação de retinol palmitato sobre os níveis de vitamina A no leite de puérperas saudáveis

The tendency towards reduction of serum retinol levels, an existing placental barrier and the increase of retinol demand, are factors that place puerperal and lactating women at risk for Vitamin A deficiency. This micronutrient is an essential component of vital processes such as differentiation, cellular proliferation, and apoptosis. The objective of this study is to evaluate the effect of palmitate retinol supplementation (100.000UI) upon the milk retinollevels in puerperal women at the Januário Cicco University Maternity Hospital. This intervention has been adopted by the Ministry of Health since 2002. The longitudinal experiment was conducted with 106 puerperal women (68 comprised the supplemented group and 38 the control group). The High Performance Liquid Chromatography (HPLC) method was used to dose the retinol of the milk and serum samples, and the creamtocrit method to determine the milk fat levels. The retinol means for the colostrums were 99.0 ± 64.4 ug/dL and 160.1 ± 94,4 ug/dl 6 hours afier supplementation; 68.9 ± 33.5 ug/dL for the transitional milk, and 30.6 ± 15.2 ug/dL for the mature milk of the supplemented group. Ali the difterences between means were statistically significant. The difterence between retinol means in the control group were also significant, with these being greater in the colostrum, 88.6 ± 62.1 ug/dL with 61.9 ± 30.1 ug/dl in the transition milk and 32.9 ±32.9 ± 17.6 ug/dL in the mature milk. No significant difference was observed in the retinol means of the three types ot milk in the supplemented group when compared to their respective means in the control group. The prevalence in serum (35.1 % and 81.1 % for the cutting point 20 ug/dL, respectively) and in milk (51.4%) revealed vitamin A deficiency as a public health problem. COlostrum, transition, and mature milk tats varied similarly in the supplemented group (1,92 ± 0,96; 3,25 ± 1,27 and 3,31 ± 1,36 grams) and in the control group (1,87 ± 1,14; 3,25 ± 1,31 and 3,36 ± 1,67 grams), with an observed difference between the colostrum/transition milk and the colostrum/mature milk fats. No difference was observed between the groups. The study showed that the 200.000UI supplementation was not sufficient to increase the milk retinol to the desired levels nor to meet the demands of the mothers with deprived hepatic reserves. It is suggested that another similar dose be offered within 30 days or less, and within 2 months post-partum, while continual/y monitoring for possible pregnancy

Otimização evalidação de métodos analíticos para a determinação simultânea de tuberculostáticos (4-FDC) por CLAE/DAD e CLUE/ DAD

Tuberculosis is a serious disease, but curable in practically 100% of new cases, since complied the principles of modern chemotherapy. Isoniazid (ISN), Rifampicin (RIF), Pyrazinamide (PYR) and Chloride Ethambutol (ETA) are considered first line drugs in the treatment of tuberculosis, by combining the highest level of efficiency with acceptable degree of toxicity. Concerning USP 33 - NF28 (2010) the chromatography analysis to 3 of 4 drugs (ISN, PYR and RIF) last in average 15 minutes and 10 minutes more to obtain the 4th drug (ETA) using a column and mobile phase mixture different, becoming its industrial application unfavorable. Thus, many studies have being carried out to minimize this problem. An alternative would use the UFLC, which is based with the same principles of HPLC, however it uses stationary phases with particles smaller than 2 μm. Therefore, this study goals to develop and validate new analytical methods to determine simultaneously the drugs by HPLC/DAD and UFLC/DAD. For this, a analytical screening was carried out, which verified that is necessary a gradient of mobile phase system A (acetate buffer:methanol 94:6 v/v) and B (acetate buffer:acetonitrile 55:45 v/v). Furthermore, to the development and optimization of the method in HPLC and UFLC, with achievement of the values of system suitability into the criteria limits required for both techniques, the validations have began. Standard solutions and tablets test solutions were prepared and injected into HPLC and UFLC, containing 0.008 mg/mL ISN, 0.043 mg/mL PYR, 0.030 mg.mL-1 ETA and 0.016 mg/mL RIF. The validation of analytical methods for HPLC and UFLC was carried out with the determination of specificity/selectivity, analytical curve, linearity, precision, limits of detection and quantification, accuracy and robustness. The methods were adequate for determination of 4 drugs separately without interfered with the others. Precise, due to the fact of the methods demonstrated since with the days variation, besides the repeatability, the values were into the level required by the regular agency. Linear (R> 0,99), once the methods were capable to demonstrate results directly proportional to the concentration of the analyte sample, within of specified range. Accurate, once the methods were capable to present values of variation coefficient and recovery percentage into the required limits (98 to 102%). The methods showed LOD and LOQ very low showing the high sensitivity of the methods for the four drugs. The robustness of the methods were evaluate, facing the temperature and flow changes, where they showed robustness just with the preview conditions established of temperature and flow, abrupt changes may influence with the results of methods

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 um reator fotoquímico aplicável no tratamento de efluentes fenólicos presentes na indústria do petróleo

With water pollution increment at the last years, so many progresses in researches about treatment of contaminated waters have been developed. In wastewaters containing highly toxic organic compounds, which the biological treatment cannot be applied, the Advanced Oxidation Processes (AOP) is an alternative for degradation of nonbiodegradable and toxic organic substances, because theses processes are generation of hydroxyl radical based on, a highly reactivate substance, with ability to degradate practically all classes of organic compounds. In general, the AOP request use of special ultraviolet (UV) lamps into the reactors. These lamps present a high electric power demand, consisting one of the largest problems for the application of these processes in industrial scale. This work involves the development of a new photochemistry reactor composed of 12 low cost black light fluorescent lamps (SYLVANIA, black light, 40 W) as UV radiation source. The studied process was the photo-Fenton system, a combination of ferrous ions, hydrogen peroxide, and UV radiation, it has been employed for the degradation of a synthetic wastewater containing phenol as pollutant model, one of the main pollutants in the petroleum industry. Preliminary experiments were carrier on to estimate operational conditions of the reactor, besides the effects of the intensity of radiation source and lamp distribution into the reactor. Samples were collected during the experiments and analyzed for determining to dissolved organic carbon (DOC) content, using a TOC analyzer Shimadzu 5000A. The High Performance Liquid Chromatography (HPLC) was also used for identification of the cathecol and hydroquinone formed during the degradation process of the phenol. The actinometry indicated 9,06⋅1018 foton⋅s-1 of photons flow, for 12 actived lamps. A factorial experimental design was elaborated which it was possible to evaluate the influence of the reactants concentration (Fe2+ and H2O2) and to determine the most favorable experimental conditions ([Fe2+] = 1,6 mM and [H2O2] = 150,5 mM). It was verified the increase of ferrous ions concentration is favorable to process until reaching a limit when the increase of ferrous ions presents a negative effect. The H2O2 exhibited a positive effect, however, in high concentrations, reaching a maximum ratio degradation. The mathematical modeling of the process was accomplished using the artificial neural network technique

Avaliação físico-química e funcional da algaroba prosopis juliflora proveniente da mesorregião agreste do Rio Grande do Norte

Algaroba (Prosopis juliflora) is a typical legume from arid and semi arid regions, which is composed by sugar-rich pods and high protein seeds. Phenolic compounds are secondary metabolites recognized as potent bioactive compounds, found in several vegetables.Therefore, the objective of this work is to characterize the algaroba flour in terms of its physicalchemical composition, total phenolic content, antioxidant activity by DPPH and ABTS methods, a-amylase and a-glycosidase inhibition, as well as to analyze its organic compounds by high performance liquid chromatography (HPLC). Three experimental groups were investigated (seeds, seeds and pod together and only pod), which were prepared by oven drying and posterior grinding. Water and ethanol extracts (70, 80, 100% v/v) were prepared and used for functional studies. Organic compounds were detected by using HPLC equipment coupled to mass spectrometer. Results show important physical-chemical differences among the experimental groups, seeds, seeds and pod together and only pod. The algarroba seed flour is high in protein (49.49%) and fat (3.10%), while the pod flour is especially rich in sugar (60.3% to 67.9%). Algaroba phenolics are concentrated in pod flour, mainly in water extracts (1.30 mg GAEQ/100g sample). All seed extracts showed high DPPH activity and maximum antioxidant activity was registered for ethanol 80% extracts (19.81 μM Trolox/g sample). The ABTS activity ranged from 9.73 to 12.74 μM Trolox/g sample. Nearly all the extracts were able to inhibit α-amylase activity mildly (30.50% to 48.80%), while the maximum α-glycosidase inhibition was observed for pod water extracts (81.03%). Algaroba water extracts proven to be especially rich in organic compounds, observed by the high number of chromatographic peaks. Results demonstrate that algaroba is a potential candidate for further investigations concerning its possible functional applications

Extração, caracterização e avaliação bioativa do extrato de Rumex acetosa

Studies show the great influence of free radicals and other oxidants as responsible for aging and degenerative diseases. On the other hand, the natural phenolic compounds has shown great as antioxidants to inhibit lipid peroxidation and lipoxygenase in vitro. Among these, is highlighted trans-resveratrol ( 3,5,4 `- trihydroxystilbene ) phenolic compound , characterized as a polyphenol stilbene class. The vegetables popularly known as "Azedinha" (Rumex Acetosa) has trans-resveratrol in its composition and from this, the present work aimed to study on the supercritical extraction and conventional extraction (Soxhlet and sequential) in roots of Rumex Acetosa, evaluating the efficiency of extractive processes, antioxidant activity, total phenolic content and quantification of trans-resveratrol contained in the extracts. Extractions using supercritical CO2 as solvent, addition of co-solvent (ethanol) and were conducted by the dynamic method in a fixed bed extractor. The trial met a 23 factorial design with three replications at the central point, with the variable reply process yield and concentration of trans-resveratrol and pressure as independent variables, temperature and concentration of co-solvent (% v/v). Yields ( mass of dry extract / mass of raw material used ) obtained from the supercritical extraction ranged from 0,8 to 7,63 % , and the best result was obtained at 250 bar and 90 °C using the co-solvent 15% ethanol (% v/v). The value was calculated for YCER a flow rate of 1,0 ± 0,17 g/min resulting in 0,0469 CO2 ( g solute / g solvent ). The results of the mass yield varied between conventional extractions 0,78 % ( hexane) and 9,97 % (ethanol). The statistical model generated from the data of the concentration of trans-resveratrol performed as meaningful and predictive for a 95% confidence. GC analysis on HPLC (High Performance Liquid Chromatography), transresveratrol was quantified in all extracts and concentration values ranged between 0,0033 and 0,42 ( mg / g extract) for supercritical extracts and between 0,449 and 17,046 (mg / g extract) to conventional extractions and therefore, the Soxhlet extraction with ethanol for more selective trans-resveratrol than the supercritical fluid. Evaluation of antioxidant (radical method to sequester 2,2- diphenyl-1- picryl - hydrazyl - DPPH) the supercritical extracts resulted in EC50 values (concentration effective to neutralize 50% of free radicals) of between 7,89 and 18,43 mg/mL , while resulting in a Soxhlet extraction with EC50 values in the range of 6,05 and 7,39 mg/mL. As for quantification of the phenolic compounds (Method Spectrophotometer Folin-Ciocalteau) the supercritical extracts resulted in values between 85,3 and 194,79 mg GAE / g extract, whereas values derived from the Soxhlet extract resulted in values between 178,5 and 237,8 mg GAE / g extract. The high antioxidant activity can not be attributed solely to the presence of phenolic compounds, but the presence of other antioxidants in the existing Rumex acetosa