Comparison between titrimetric and spectrophotometric methods for quantification of vitamin C.

Vitamin C is usually quantified by titrimetric or chromatographic methods. However, these methods have limitations: food color interferes with the titrimetric method and the chromatographic method is costly. The aim of this study was to compare a spectrophotometric method, based on reduction of cupric ions in the presence of cuproine complex, with a titrimetric method, based on reduction of 2,6-dicholorophenolindophenol. Linearity, precision, accuracy, and limits of detection (LOD) and quantification (LOQ) were evaluated using a standard vitamin C solution. Both methods were also applied for AA quantification in industrialized orange and pineapple juices. The methods were precise and accurate when applied to the standard solution. The spectrophotometric method was more sensitive, with lower values for LOD (0.002mgmL-1) and LOQ (0.010mgmL-1), and more accurate with error less than 5% while results from the titrimetric method were affected by the juice color, which generated errors in excess of 15%.

Studies on lipase-affinity adsorption using response-surface analysis.

Lipases are widely distributed enzymes that can be obtained from animals, plants and micro-organisms. Coupling lipases with a wide range of substrates allows the opportunity for synthesis of optically pure pharmaceutical preparations, flavour compounds and other food additives. Affinity chromatography owes its power as a purification method to specific biological interactions. Response-surface analysis was chosen to study column efficiency. This method allows the understanding of interactions between variables with advantages over conventional methods, which involve changing one variable while fixing others at certain levels. The aim of this work was to study the influence of the ratio bed height/column diameter (L/D) and superficial velocity (V) on the column efficiency. The experimental design involved the two variables, L/D (2-10) and v (1-2 cm/min), at five levels. Lipase was obtained from Geotrichum sp. culture in a complex medium composed of 5% corn-steep liquor, 0.5% NH(4)NO(3) and 1% olive oil at 30 degrees C, with 1VVM (air volume/medium volume per min) aeration and 400 rev./min agitation. Maximum lipase activity was 19 units/ml after almost 9 h of fermentation. This lipase could potentially be used in esterification reactions to increase the content of gamma-linolenic acid and to produce bioaromas for food industries. The adsorption assays were carried out in a fixed-bed column with an affinity adsorbent, which was obtained by reaction of a gel with oleic acid as ligand. Breakthrough curves were obtained for all experiments. It has been shown that the lower the values of both L/D and v, the higher the column efficiency (maximum 65.43%). Also, it was observed from the response surface that the efficiency reached a minimum at an L/D of around 8.

New rat model for attention deficit hyperactive disorder (ADHD).

PURPOSE: In a strain of the Long-Evans Cinnamon (LEC) rats, we found spontaneously hyperactive animals designated as "wiggling," and established a congenic wiggling (Wig) rat by transferring the gene from the LEC to the Wistar King-Aptekman/Hokkaido (WKAH) strain. We evaluated the feasibility of the Wig rat for an animal model of human attention deficit hyperactive disorder (ADHD). METHODS: Mode of inheritance was examined by use of linkage analyses. Motor activity, behavior, and working memory were assessed by use of electric digital counters, open field test, and Y-maze and water-maze tests. RESULTS: The abnormal behavior, including hyperactivity, was transmitted in autosomal recessive mode. Diurnal and nocturnal motor activity of 12- to 14-week-old Wig rats was markedly higher than that of controls, and this hyperactivity was more prominent during nighttime than daytime. Ambulation in the open-field test was significantly increased in Wig rats, but rearing was decreased in Wig rats, compared with controls. Results of the Y-maze tests indicated that spontaneous alternation behavior was significantly impaired in Wig rats, although there was no difference in the total arm entries. The water-maze test could not be performed because, when exposed to water, Wig rats panicked and almost drowned. CONCLUSIONS: Wig rats are hyperactive and have impaired working memory and impulsive behavior, as assessed by the motor activity and open-field tests and the Y-maze test, and these abnormalities are transmitted by a single gene with Mendelian pattern. Wig rats represent an excellent animal model of human ADHD.

Effects of sex hormones on fulminant hepatitis in LEC rats: a model of Wilson's disease.

LEC rats, which have hereditary hepatitis and have recently been proposed as an animal model for Wilson's disease, were examined to determine the effects of sex hormones on fulminant hepatitis. After the rats had undergone ovariectomies or orchidectomies (castration) and were compared with intact rats, the age at the onset of fulminant hepatitis was not substantially altered but the survival rates decreased from 50% to 12.5% for females and 75% to 14.3% for males, indicating that sex hormones did not influence the occurrence of fulminant hepatitis but influenced mortality due to fulminant hepatitis. When testosterone was administered to the ovariectomized or orchidectomized rats, the survival rate increased to over 90% in both sexes. In contrast, estradiol did not affect the survival rate of either sex but affected the onset of fulminant hepatitis. That is, with the administration of estradiol, the age at which serum GPT activity reached its maximum was delayed 4 weeks in ovariectomized rats and 6 weeks in orchidectomized rats as compared with intact rats. A similar but somewhat weaker tendency appeared in rats given progesterone. The results of our study indicate that sex hormones have no effect on the rate of occurrence of hepatitis but affect the progression of hepatitis. In particular, testosterone increased the survival rate of rats with fulminant hepatitis, and exogenous estradiol delayed the onset of hepatitis for several weeks.

Clinico-pathological studies of LEC rats with hereditary hepatitis and hepatoma in the acute phase of hepatitis.

The LEC rat, which suffers from hereditary hepatitis, was examined for elucidation of its clinicopathological characteristics during development of the acute phase of hepatitis by quantitative analyses of histological observations of the liver in combination with laboratory data on various serum enzymes. The progression of acute hepatitis in the LEC rat was observed to begin insidiously early in life, i.e., a few enlarged hepatocytes and Councilman bodies appeared at around 8 weeks of age without clinical signs. Furthermore, it was revealed that the acute phase of hepatitis started with a remarkable increase of Councilman bodies, large nuclei and hepatocytes in mitosis in the liver 3 to 4 weeks before the onset of fulminant hepatitis, which is characterized by the elevation of serum enzyme activities such as GOT, GPT and gamma-GTP, and the onset of jaundice. From those observations, three stages were proposed for the progression of acute hepatitis in the LEC rat.