Liquid chromatographic determination of carminic acid in yogurt

A reverse-phase liquid chromatographic method is described for the determination of carminic acid in yogurt. A C18 column is used with acetonitrile-1.19M formic acid (19 + 81) as mobile phase and diode array detection. Sample preparation includes deproteinization with papain and purification in a polyamide column. The relative standard deviation for repeated determinations of carminic acid in a commercial strawberry-flavored yogurt was 3.0%. Recoveries of carminic acid added to a natural-flavored yogurt ranged from 87.2 to 95.3% with a mean of 90.2%. The method permits measurement of amounts as low as 0.10 mg/kg.     

Quantitative determination of L-arginine by enzymatic end-point analysis

An enzymatic end-point method for the quantitative determination of L-arginine was evaluated with samples of synthetic wine and natural grape juice. The enzymes arginase, urease, and glutamate dehydrogenase were used in this simple assay, similar to those described for many metabolites by Boehringer-Mannheim. In synthetic wine, recovery of L-arginine ranged between 98.3 and 104.4% and the precision as coefficient of variation was between 0.4 and 1.47% in the concentration range of the method, 0-100 mg/L L-arginine. The recovery of L-arginine in a grape juice with added L-arginine after clarification with polyvinylpolypyrrolidone ranged between 100 and 101.3%, and the coefficient of variation was 0.6%. The method has low material costs of approximately 0.43 U.S.$ per assay, and the time course of the reaction facilitates measurement of several samples concurrently. The results of this evaluation indicate that the enzymatic assay is a preferred method over colorimetric methods for the manual determination of L-arginine.     

Capillary gas chromatography with atomic emission detection for pesticide analysis in soil samples

A method for the simultaneous determination of 10 pesticides (organochlorines, organophosphorus compounds, and pyrethrins) in soils using capillary gas chromatography with atomic emission detection (GC-AED) is reported. Soil samples are first "cleaned-up" with 25 mL of an ascorbic acid solution (pH 2.15). The aqueous phase is extracted with ethyl acetate, and the solid residue is then extracted twice with 10 mL of ethyl acetate. The three resultant organic extracts are combined, concentrated to dryness, and reconstituted in 1 mL of acetone. The pesticides are selectively detected by monitoring chlorine and bromine in the first run and sulfur emission line wavelength in the second run. Each chromatographic run takes 19 min. Detection limits are between 25 and 75 pg, depending on the compound, which corresponds to 1.7 and 5.0 ng/g in the soil samples, respectively. Recoveries of the pesticides from spiked preparations result in an overall mean recovery of 95.3% (n = 120) at fortification levels ranging from 10 to 60 ng/g, depending on the compound. The method is reliable and can be useful for routine monitoring in soils.     

Immunochemical determination of 2,4,6-trichloroanisole as the responsible agent for the musty odor in foods. 2. Immunoassay evaluation

Immunoassays for 2,4,6-trichloroanisol (TCA) have been evaluated. The assays were developed after raising antibodies against three different immunizing haptens (1). Lack of reproducibility has been one of the main problems of these assays. Precision was worse on these assays, reaching lower limits of detection. The high lipophilicity of TCA and its, consequently, low water solubility have been found to be the major cause of this problem. A reliable microplate-based enzyme-linked immunosorbent assay (ELISA) has been set after consideration of the TCA physicochemical features and evaluation of important parameters affecting immunoassay performance. The immunoassay uses As78 (developed against hapten B-KLH) and C9-OVA as the coating antigen. The selectivity is high although the brominated analogue 2,4,6-TBA is also recognized. In buffered media containing 7% ethanol, the resulting assay shows a good accuracy with an IC(50) value of 0.53 microgram L(-)(1) and a limit of detection of 0.044 microgram L(-)(1). Red and white wine samples caused important interferences in the immunoassay demonstrating the necessity of a cleanup procedure prior to the ELISA.     

Molecular detection and quantification of Colletotrichum abscissum in sweet orange propagative material

Postbloom fruit drop disease (PFD) has caused serious impacts on citrus production in the Americas, occurring sporadically and suddenly during rain in the flowering period. In São Paulo State, Brazil, Colletotrichum abscissum is the causal agent responsible for over 80% of the disease incidence. Pathogen dispersal over long distances and the origin of primary inoculum are still unclear for PFD epidemics. We tested the hypothesis that citrus propagation material can harbour C. abscissum DNA by quantifying it in leaves of budwood increase block trees (BIB) and young citrus plants (YP) using multiplex quantitative PCR (qPCR). C. abscissum DNA was detected in all citrus nurseries, regardless of the type of propagative material, the sweet orange variety, or the nursery location. Overall, 73.4% of all samples from citrus nurseries have DNA from the pathogen, with a detection limit of 10 conidia. The average of 155 conidia found in YP was higher than the conidia observed on leaf samples from BIB (p = 0.03), although leaf samples from cultivars Valencia and Pera did not differ significantly, with means around 127 and 118 conidia, respectively (p = 0.75). This is the first molecular detection of C. abscissum in citrus propagative material. The multiplex qPCR assay may be used as a protocol for an accurate diagnosis of C. abscissum in citrus propagative material, may assist a better understanding of the pathogen dispersal over long distances, and may be used for further studies involving the quantification of C. abscissum in citrus orchards.     

Prolonged Waterlogging Reduces Growth and Yield in Broccoli Plants (Brassica oleracea var. italica)

The crop of broccoli in tropical regions is of great importance among flowering vegetables; however, the yield of this crop is severely impacted by climatic variations that can cause floods. In Tunja, Colombia, a study was carried out under greenhouse conditions in which the tolerance of broccoli plants to prolonged waterlogging was evaluated. One group of plants were kept under waterlogging conditions until most of them showed severe symptoms of chlorosis while another group was grown under regularly drained and watered soil conditions as a control. Waterlogging caused the death of 20% of the plants, reduced the height of the plants by 42.9%, the thickness of the stem by 42.1%, the foliar area by 87%, the chlorophyll content in the leaves by 96.6%, and the total dry weight per plant by 79.9%. The absolute and relative growth rates decreased by 80 and 24.4%, respectively. Waterlogging also prevented flower production and caused a 23.7% increase in the accumulation of biomass in roots but reduced it by 24.5% in leaves. Likewise, the net assimilation rate fell 72.3% when waterlogged and the values of allometric variables which express growth were altered by this stressor. Consequently, it can be inferred that these plants have a low tolerance to waterlogging; however, the most severe impact caused by waterlogging was the inability of plants to develop flowers. The lack of flowers is devastating due to their economic and commercial importance of broccoli, and they are the primary justification for the cultivation of these plants.

     

NITROGEN ACCUMULATION IN SHOOTS AS A FUNCTION OF GROWTH STAGE OF CORN AND WINTER WHEAT

Midseason fertilizer nitrogen (N) rates based on predicted yields can be projected if the quantity of N accumulated in winter wheat (Triticum aestivum L.) and corn (Zea mays L.) is known especially early in the growing season. This study was conducted in 2006 and 2007 to establish the amount of N accumulated in corn and winter wheat over the entire growing season. Plots representing three N fertilization rates 0, 45, and 90 kg ha^?1 at Stillwater and 0, 67, and 112 kg ha^?1 at Lahoma were selected from two long-term wheat experiments located at research stations in Stillwater and Lahoma, Oklahoma. For corn, three N fertilization rates 0, 112 and 224 kg ha^?1 at Lake Carl Blackwell and 0, 56 and 112 kg ha^?1 at Perkins were selected from N studies, located at research stations near Lake Carl Blackwell and Perkins, Oklahoma. Sequential aboveground biomass samples were collected from 1 m² area of wheat and 1.5 m long row (0.76 cm spacing) for corn throughout their respective growing seasons. In general, this work showed that more than 45% of the maximum total N accumulated could be found in corn plants by growth stage V8 (8th leaf collar fully unfolded). For winter wheat, more than 61% of the maximum total N accumulated at later stages of growth could be accounted for by Feekes growth stage 5 (F5, leaf strongly erected). Our findings are consistent with those of others showing that yield potential can be predicted at mid-season since such a large percentage of the total N accumulated was accounted for early on in the growing cycle of either wheat or corn.     

Salinity effects on growth analysis and nutrient composition in four grain legumes‐rhizobium symbiosis

The effect of salinity on growth response, nitrogen (N) fixation and tissue mineral content was investigated for four legumes: faba bean (Vicia faba L), pea (Pisum sativum L), soybean (Glycine max L), and common bean (Phaseolus vulgaris L). Plants were grown in a vermiculite culture system supplied with a N‐free nutrient solution with the addition of 0, 50, and 100 mM NaCl. Plants were harvested at the beginning of the flowering period and the dry weights of shoots and roots and acetylene reduction activity (ARA) were evaluated at the same time plant tissues were analysed for N, potassium (K), calcium (Ca), magnesium (Mg), and sodium (Na) contents.

The depressive effect of saline stress on ARA of nodules was directely related to the salt induced decline in dry weight and N content in shoots. Growth inhibition by NaCl treatments was greater for the pea than for other legumes, whereas the soybean was the most salt‐tolerant Saline stress also affected the N content in shoots and roots. In general the N content accumulated in the shoot and Na in the roots of the four legumes tested, while K accumulated both organs. The acquisition of other macronutrients differed according to the legume species. The legumes most sensitive were P. sativum and V. faba which accumulated Ca in shoot and Mg both in the shoot and the roots. On the contrary, in G. max and P. vulgaris, the two most salt tolerant legumes, accumulated Mg in the roots and Ca in both vegetative organs. Our results suggest a relationship between the salt‐tolerant range in legumes and the macronutrient accumulation in vegetative organs.