Validation of an immunoassay method for the determination of traces of carbaryl in vegetable and fruit extracts by liquid chromatography with photodiode array and mass spectrometric detection

A competitive enzyme-linked immunosorbent assay (ELISA) method for carbaryl quantitation in crop extracts was validated by liquid chromatography (LC) with diode array detection (DAD). For this purpose, six crops (banana, carrot, green bean, orange, peach and potato) were chosen for recovery and reproducibility studies. The general sample preparation included extraction with methanol followed by liquid-liquid partitioning and clean-up on Celite-charcoal adsorbent column of the vegetable extracts. ELISA samples consisted of a diluted LC extract in assay phosphate buffer (pH 7.5). The potential effect of methanol in these samples was evaluated. It was observed that a maximum content of 10% methanol present in the assay buffer could be tolerated without expressive losses in the ELISA performance. Under these conditions, a IC50 similar to 1.48 mu g l(-1) was obtained. A minimum matrix effect with a 1:50 dilution of the methanolic extracts in assay buffer was noticed, except for green bean samples that inhibited completely the assay. For the vegetable extracts, the ELISA sensitivities varied from 3.9 to 5.7 mu g l(-1), and good recoveries (82-96%) with R.S.D.s ranging from 5.7 to 12.1% were found. An excellent correlation between the LC-DAD and ELISA techniques was obtained. The confirmation of the carbaryl in less concentrated samples was achieved by LC-mass spectrometry interfaced with atmospheric pressure chemical ionisation. The [M + H](+)= 202 and [M + H-57](+)=145 ions, equivalent to the protonated molecular and l-naphthol ions, respectively, were used to carbaryl identification in these samples. (C) 1998 Elsevier B.V. B.V. All rights reserved.

Validation of a stability-indicating HPLC method with diode array detection for the determination of beclomethasone dipropionate in aqueous suspension for nebulizer

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Chemical and Antidiarrheal Studies of Plinia cauliflora

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

Enrofloxacin assay validation and pharmacokinetics following a single oral dose in chickens

The pharmacokinetics of enrofloxacin (ENRO), a fluoroquinolone antimicrobial agent, was studied in male broiler chickens (Cobb) after single oral administration of 10 mg of ENRO/kg b.w. A high-performance liquid chromatography-photodiode array detector (DAD) (HPLC-DAD) method was developed and validated and used for quantitation of ENRO and its major metabolite ciprofloxacin in plasma. The HPLC analyses were carried out using a cationic-octadecyl mixed column and 0.05 mol/L phosphate buffer (pH 2.5)/acetonitrile as mobile phase. The sample preparation of plasma consisted of the precipitation of proteins followed by solid phase extraction on cationic-octadecyl mixed cartridges. The method was validated considering linear range, linearity, selectivity, sensitivity, limit of detection (LOD), limit of quantitation (LOQ), intra- and inter-day precisions and accuracy. The LOD and LOQ for both fluoroquinolones were 60 and 200 ng/mL for plasma. The plasma concentration vs. time graph was characteristic of a two-compartment open model. The maximal plasma concentration of 1.5 +/- 0.2 mg/mL was achieved at 9 +/- 2 h. The elimination half-life and the mean residence time of ENRO were 1.5 +/- 0.2 and 15.64 h, respectively. The area under the concentration-time curve was calculated as 35 +/- 4 mg(.)h/mL.

Determinação de enantiômeros em extratos vegetais por cromatografia quiral e dicroísmo circular

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Continuous vs. segmented second-dimension system gradients for comprehensive two-dimensional liquid chromatography of sugarcane (Saccharum spp.)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Flavonoid Detection in Hydroethanolic Extract of Pouteria torta (Sapotaceae) Leaves by HPLC-DAD and the Determination of Its Mutagenic Activity

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

Acetone as a greener alternative to acetonitrile in liquid chromatographic fingerprinting

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

Determinação do teor dos ingredientes ativos imidacloprida e clorpirifós em produtos técnicos formulados por CLUE-DAD

The use of pesticides to manage pest problems in agriculture has become a common practice around the world. Pesticides consist in chemical substances or biological agents that act preventing, destroying, repelling or mitigating the damage caused by pests in agriculture. In order to a successful treatment, the correct dosage of pesticide has to be applied for each kind of culture. In this context, this project aims to quantify the levels of the active ingredients Imidacloprid and Chlorpyrifos present in formulated technical products and compare with the reported in the ir commercial leaflets. The analyzes were performed by Ultra Performance Liquid Chromatography coupled to a photodiode array detector (CLUE-DAD). The identification of Imidacloprid and Chlorpyrifos in the samples was realized by comparison of the time of retention and UV spectre of those substances with the commercials patterns, resulting in a excellent method specificity of the method. In the scope of determination of active ingredients in formulated products, the method developed showed good sensitivity in the applied conditions, with suitables limits of detection (4,2.10-² mg L-¹ for Imidacloprid and 1,1.10-¹ mg L-¹ to Chlorpyrifos) and quantitation (1,4.10-¹ mg L-¹ and Imidacloprid 3,6.10-¹ mg L-¹ to Chlorpyrifos) for the concentrations found in matrices analyzed. Furthermore, good results were obtained to the method accuracy with coeficiente of variance values in the range of 0,6 to 0,7% for repeatability (intra-day) and 0,4 to 0,58% for intermediate precision (inter- day). The accuracy of the method was determined by recovery tests, the values were comprised in the range of 98 102%. According to the recommendations of the Associação Brasileira de Normas Técnicas, these values show good accuracy of the method. The concentrations obtained in the quantification of the active ingredients in formulated products showed values within permited by ABNT, 60.1% (w/v) ± 0.6%...

An LC-DAD Fingerprinting Method for Alkaloids, Flavonoids and Styrylpyrones from Cryptocarya mandioccana

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

Secondary Metabolites of Miconia rubiginosa

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

Rapid degradation of propanil in rice crop fields

Propanil and its major degradation product, 3,4-dichloroaniline (DCA), were monitored in surface water and soil samples from two rice fields of the Ebre Delta area (Tarragona, Spain) following agricultural application. On-line solid-phase extraction (SPE) (water) and Soxhlet extraction (soil) followed by liquid chromatography/diode array detection (LC/DAD) were used for the trace determination of both compounds. Unequivocal confirmation/identification was conducted by using liquid chromatography/atmospheric pressure chemical ionization-mass spectrometry, LC/APCI/MS (using negative and positive ionization modes). Concentrations of the herbicide propanil in water samples varied from 1.9 to 55.9 mu g/L. Propanil degraded very rapidly to DCA, and high concentrations of this product were found, varying from 16.5 to 470 mu g/L in water and 119 +/- 22 mu g/kg in soil samples. No detectable DCA (<0.001%) was found in the applied pesticide formulation, indicating that DCA formation took place after propanil application. These field results compared favorably with laboratory experiments showing that humic interactions had a strong influence on the pesticide degradation. The half-lifes under real conditions for propanil and DCA, calculated using a first-order decay, were 1.2 and 1.6 days, respectively.

Multiresidue analysis of pesticides in water from rice cultures by on-line solid phase extraction followed by LC-DAD

An automated on-line solid phase extraction procedure followed by liquid chromatography with diode array detection was investigated for the determination of different classes of pesticides in water samples containing varied amount of humic substances. The different pesticides used were: carbendazin, carbofuran, atrazine, diuron, propanil, molinate, alachlor, parathion-ethyl, diazinon, trifluralin and the degradation products deisopropylatrazine and deethylatrazine. Humic substances extracted from a Brazilian sediment were used from 5 to 80 mg/l and their influence on recoveries was evaluated in neutral and acidic media. Recoveries higher than 70% were obtained for all the pesticides, from the preconcentration of 75 mi of aqueous sample fortified at 2 ng/ml using precolumns packed with PLRP-S. Good recoveries were obtained at neutral pH for most of the analytes up to 40 mg/l of humic acid. Only at 80 mg/l the recoveries were significantly affected, both at acidic and neutral pH. The method was applied to the determination of pesticides in river water spiked at 0.1 to 1 ng/ml. Detection limits obtained for water containing 10 mg/l of humic acid were between 0.05 and 0.3 ng/ml.

LC-DAD Determination of Fleroxacin in Bulk and Pharmaceutical Dosage Forms

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Development of an analytical method for the quantification of pfaffic acid in Brazilian ginseng (Hebanthe eriantha)

Hebanthe eriantha (Poir.) Pedersen (Amaranthaceae), which is known as Brazilian ginseng is widely used in folk medicine as an aphrodisiac and antidiabetic tonic. The anti-tumor activity, attributed to the pfaffic acid present in roots of H. eriantha, is responsible for the great interest in the commercialization of this species. In Brazil, the species H. eriantha is mainly used in commercial preparations, although other plants of the genus Pfaffia and Hebanthe have been marketed as Pfaffia paniculata or Brazilian ginseng. The pfaffic acid present in the roots is mainly conjugated with sugars (pfaffosides) and can be used as an active marker of H. eriantha, which helps to differentiate this species from others marketed as Brazilian ginseng. The main objective of this study was to develop and validate a liquid chromatographic method to quantify pfaffic acid in the roots of H. eriantha. The extraction and hydrolysis conditions were optimized using an univariate and experimental design, respectively, and the quantification of pfaffic acid by high performance liquid chromatography with diode-array detection (HPLC-DAD) was validated. This method was used to evaluate the pfaffic acid content in 30 different genotypes of the species from a germplasm collection. The content of pfaffic acid ranged from 0.97 to 4.29% (w/w) on a dry weight basis. © 2013 Elsevier B.V.