Development of practical HPLC methods for the separation and determination of eggplant steroidal glycoalkaloids and their aglycones

A practical set of HPLC methods was developed for the separation and determination of the eggplant steroidal glycoalkaloids, solanine, chaconine, solasonine, solamargine, and their aglycones, solasodine and solanidine. A gradient method was initially developed, but proved to be neither robust nor practical. Three separate isocratic methods using acetonitrile and ammonium dihydrogen phosphate were developed and shown to be more repeatable, less subject to fluctuations in mobile phase composition, and less time consuming. The effect of adjusting buffer pH, column temperature, and buffer type (triethylammonium phosphate vs. ammonium dihydrogen phosphate) were evaluated. It was also discovered that, by addition of 10% methanol to the acetonitrile portion of the mobile phase, more control over the separations was possible. The use of methanol as a mobile phase entrainer greatly improved separations in some cases and its effectiveness was also dependent upon column temperature. Assessments of the method recovery, limit of detection, and limit of quantitation were made using extracts from S. melongena and S. linnaeanum.

Effect of cocaine on periadolescent rats with or without early maternal separation

Cocaine-induced behavioral sensitization and weight loss were investigated in periadolescent Wistar rats kept with their mothers or subjected to repeated maternal separation. Litters allocated to the separation procedure were placed in a temperature-controlled (33ºC) chamber for 3 h per day from postnatal day 6 (P6) to P20. Non-handled rats were left undisturbed until weaning. Treatments were started on P30-31 and the test was performed on P36-37. Animals received injections of saline or cocaine (10 mg/kg, sc) twice daily for 5 days. on day 6 all animals received saline. on day 7 animals were challenged with 10 mg/kg cocaine and their locomotion was evaluated in activity cages. A third group received saline throughout the 7-day period. Body weights were recorded on P30-31 and P36-37. Two-way ANOVA on body weights showed a main effect of treatment group (F(1,35) = 10.446, P = 0.003; N = 10-12). Non-handled rats treated with cocaine for 5 days gained significantly less weight, while no significant effect was observed in maternally separated rats. Two-way ANOVA revealed a main effect of drug treatment on locomotor activity (F(2,32) = 15.209, P<0.001; N = 6-8), but not on rearing condition (F(1,32)<0.001, P = 0.998). Animals pretreated with cocaine showed a clear behavioral sensitization relative to the saline group. No difference in the magnitude of sensitization was found between separated and non-handled animals. Only the effect of cocaine on weight gain was significantly affected by repeated episodes of early maternal separation during the pre-weaning period.

Maternal separation stress in male mice: long-term increases in alcohol intake

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Photoinduced hole-transfer in semiconducting polymer/low-bandgap cyanine dye blends: evidence for unit charge separation quantum yield

Power-conversion efficiencies of organic heterojunction solar cells can be increased by using semiconducting donor-acceptor materials with complementary absorption spectra extending to the near-infrared region. Here, we used continuous wave fluorescence and absorption, as well as nanosecond transient absorption spectroscopy to study the initial charge transfer step for blends of a donor poly(p-phenylenevinylene) derivative and low-band gap cyanine dyes serving as electron acceptors. Electron transfer is the dominant relaxation process after photoexcitation of the donor. Hole transfer after cyanine photoexcitation occurs with an efficiency close to unity up to dye concentrations of similar to 30 wt%. Cyanines present an efficient self-quenching mechanism of their fluorescence, and for higher dye loadings in the blend, or pure cyanine films, this process effectively reduces the hole transfer. Comparison between dye emission in an inert polystyrene matrix and the donor matrix allowed us to separate the influence of self-quenching and charge transfer mechanisms. Favorable photovoltaic bilayer performance, including high open-circuit voltages of similar to 1 V confirmed the results from optical experiments. The characteristics of solar cells using different dyes also highlighted the need for balanced adjustment of the energy levels and their offsets at the heterojunction when using low-bandgap materials, and accentuated important effects of interface interactions and solid-state packing on charge generation and transport.

Magnetic interactions and spin-reversal mechanism in ErMn1-xCoxO3-delta samples

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

On line extraction and separation of bendiocarb, methomyl, methylparathion, and pentachlorophenol pesticides from raw milk

A high performance liquid chromatography (HPLC) method for extraction and determination of pesticides from raw milk was developed. The method involves direct injection of raw milk samples on a bovine serum albumin-dimethyl-octyl-silica gel (BSA-Si-Cs) column. The mobile phase 0.05 mol.L-1 phosphate buffer pH6.0 in acetonitrile (70:30 v/v) was employed for extraction and separation of bendiocarb, methylparathion, pentachlorophenol, and methomyl pesticides. The method shows good results of recovery in the pesticides studied, higher than 99.6%.

High-performance liquid chromatographic separation and identification of polyphenolic compounds from the infusion of Davilla elliptica St. Hill

The isolation of polyphenolic compounds from an infusion of the Brazilian plant Davilla elliptica (Dilleniaceae), used as tea by virtue of its digestive properties, is described. An improved preparative HPLC method was used in order to isolate pure polyphenols from the complex mixture. Liquid-liquid extraction and solid-phase extraction were employed to minimise the interference of polymeric compounds and to provide an enriched fraction of the compounds of interest. The identification of the isolated compounds was performed using analytical HPLC as well as direct injection electrospray ionisation ion trap tandem mass spectrometry (ESI-IT-MS/MS). The high flavonoid content suggests that D. elliptica may be a promising source of compounds to produce natural phytomedicines. Copyright (C) 2007 John Wiley & Sons, Ltd.

Genetic and environmental parameters for racing time at different distances in Brazilian Thoroughbreds

The aim of the present study was to investigate genetic parameters for racing time in Thoroughbred horses racing at distances between 1000 and 1600 m subdivided into 100-m intervals. The data provided by TURFETOTAL Ltda comprised races that occurred in the Gavea and Cidade Jardim race tracks over a period of 11 years (1992-2002) and consisted of 32 145 races and 238 890 time records. The variance components necessary to obtain the heritability and repeatability estimates of the traits studied were estimated with the MTDFREML program, and animal age at race (3 years old or younger, 4, 5 and older than 5 years), sex (male and female), number of races (1-32 145), and postposition at start (1-11) as fixed effects, and animal and permanent environmental random effects were included in a one-trait animal model. Males were significantly superior to females at all distances. Excluding the 1100 m distance, animals 4 years of age were significantly faster than the mean of the other ages for all distances analysed. Horses older than 5 years showed a significantly lower performance than the mean of the other ages for all distances analysed, except for the 1100 m. Postpositions one and two did not differ significantly from one another for any of the distances analysed. These two inner positions both together varied from the other positions depending on race length. The components of additive genetic and permanent environmental variance varied in a similar way, tending to decrease with increasing racing distance, and the other temporary environmental variance almost doubled from 1000 to 1600 m. As was the case for the additive genetic and environmental variances, heritability and repeatability estimates tended to decrease with increasing distance, indicating that selection based on racing time will be less successful when the racing distance increases.

Preparative droplet counter-current chromatography for the separation of the new nor-seco-triterpene and pentacyclic triterpenoids from Qualea parviflora

The methanolic extract of the bark of the medicinal plant Qualea parviflora (Vochysiaceae) contains new nor-seco-triterpene and pentacyclic triterpenoids. They were separated in a preparative scale using droplet counter-current chromatography. The optimum solvent used was composed of a mixture of CHCl3/MeOH/H2O (43:37:20, v/v) in the descending mode and led to a successful separation of the new compound 28-nor-17, 22-seco-2 alpha, 3 beta, 19, 22, 23-pentahydroxy-Delta(12)-olecinane, besides the known triterpenoids bellericagenin B, bellericaside B and arjunglucoside I. Identification was performed by ESI-MS, H-1 NMR and C-13 NMR analyses.

Application of preparative high-speed counter-current chromatography for the separation of flavonoids from the leaves of Byrsonima crassa Niedenzu (IK)

The methanolic extract of the leaves of the medicinal plant Byrsonima crassa (Malpighiaceae) contain flavonoids with antioxidant activity. They were separated in a preparative scale using high-speed counter-current chromatography. The optimum solvent system used was composed of a mixture of ethyl acetate-n-propanol-water (140:8:80 (v/v/v)) and led to a successful separation between monoglucosilated flavonoids (quercetin-3-O-alpha-L-arabinoside, quercetin-3-O-beta-D-galactoside) and the biflavonoid amentoflavone in only 3.5 h. The purities of quercetin-3-O-alpha-L-arabinoside (95 mg), quercetin-3-O-beta-D-galactoside (16 mg) and the biflavonoid amentoflavone (114 mg) were all isolated at purity over 95%. Identification was performed by H-1 NMR, C-13 NMR and UV analyses. (C) 2004 Published by Elsevier B.V.

Metallomic study on plasma samples from Nile tilapia using SR-XRF and GFAAS after separation by 2D PAGE: initial results

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Analytical approach to the metallomic of Nile tilapia (Oreochromis niloticus) liver tissue by SRXRF and FAAS after 2D-PAGE separation: Preliminary results

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)