Collaborative validation of the QuEChERS procedure for the determination of pesticides in food by LC-MS/MS

Seven FDA pesticide laboratories collaborated to develop and validate an LC-MS/MS method to determine 173 pesticides in <20 min. The average determination coefficient (r2) was >0.99 for all but two compounds tested. The limits of detection were <20 ng/mL for all compounds and <10 ng/mL for 363 of the 368 transitions reported. The method was used to determine pesticides in two AOAC sponsored proficiency samples. The LC-MS/MS determination was used for the analysis of oranges, carrots and spinach using the QuEChERS (Quick, Easy, Cheap, Effective, Rugged, Safe) method. Each matrix was fortified at 20, 100, 400, and 1000 ng/g. No false positive responses were detected in controls of the three matrices. 165 pesticides had recoveries between 70 and 130%, and 161 had minimum detection levels less than 10 ng/g. Recoveries of 169 compounds for the 1000 ng/g spikes were within 50-150%. A matrix effect study indicated all three matrices caused a small net suppressing effect, the most pronounced attributable to the citrus matrix. The procedure proved to be accurate, precise, linear, sensitive and rugged, and adds 100 pesticides to the scope of the FDA pesticide program.     

Use of surface plasmon resonance (SPR) to study the dissociation and polysaccharide binding of casein micelles and caseins

Tests were made to determine whether surface plasmon resonance (SPR) could be used as a technique to study the dissociation properties of bovine casein micelles or of sodium caseinate and the interactions between these protein particles and different polysaccharides. Surfaces of bound micelles or caseinate were made, and the changes in refractive index of these layers were used to define changes in the structures of the chemisorbed material. The technique appears to have some potential for studying details of the dissociation of casein micelles and of the binding of different polysaccharides to caseins.     

Microbial and microfaunal community structure in cropping systems with genetically modified plants

Soils from field sites at Foulum (DK), Narbons (FR) and Varois (FR) planted with genetically modified maize expressing either the insecticidal Bacillus thuringiensis protein (Bt) or herbicide tolerance (HT), as described elsewhere in this volume, were analysed for nematodes, protozoa and microbial community structure. These analyses were mirrored in single-species testing and in mesocosm experiments, and were coordinated with field samples taken for microarthropods, enchytraeids and earthworms so allowing for cross-comparison and a better understanding of the results observed in the field. Over the first 2 years of the field experiments (in 2002 and 2003), the effect of Bt-maize was within the normal variation expected in these agricultural systems. Sampling in 2004 and 2005 was expanded to include the effects of tillage (i.e. reduced tillage versus conventional tillage) and also the use of HT-maize. Tillage had major effects regardless of soil type (Varois or Foulum), with reduced-tillage plots having a greater abundance of microfauna and a different microbial community structure (measured both by phospholipid fatty-acid analysis (PLFA) and by community-level physiological profiling (CLPP)) from conventionally tilled plots. Grass, as a contrasting cropping system to maize, also had an effect regardless of soil type and resulted in greater microfaunal abundance and an altered microbial community structure. Differences in crop management, which for the Bt-maize was removal of the insecticide used to control European corn borer and for HT-maize was a change in herbicide formulation, were only tested at single sites. There were differences in microbial community structure (CLPP but not PLFA) and sporadic increases in protozoan abundance under the Bt-crop management. The HT-maize cropping system, which covered a shorter period and only one site, showed little change from the conventional system other than an altered microbial community structure (as measured by PLFA only) at the final harvest. The Bt-trait had a minimal impact, with fewer amoebae at Foulum in May 2003, fewer nematodes at Foulum in May 2004 but more protozoa at Varois in October 2002 and an altered microbial community structure (PLFA) at Foulum in August 2005. These were not persistent effects and could not be distinguished from varietal effects. Based on the field evaluations of microfauna and microorganisms, we conclude that there were no soil ecological consequences for these communities associated with the use of Bt- or HT-maize in place of conventional varieties. Other land management options, such as tillage, crop type and pest management regime, had significantly larger effects on the biology of the soil than the type of maize grown.     

In vivo investigation of changes in biomarkers of oxidative stress induced by plant food rich diets

It is well established that vegetables and fruit (VF) contain antioxidant phytochemicals. Consequently, it is expected that individuals who consume diets with a high content of VF should be better protected against oxidative cellular damage than individuals who do not, but not all data support this assumption. The objective of this study was to identify possible explanations for this conundrum. The effects of two diets that differed in VF content on markers of oxidative damage were studied. Sixty-four women participated in a 14-day dietary intervention. Subjects consumed on average either 3.6 or 12.1 servings of VF per day. The primary end points assessed were 8-hydroxy-2-deoxyguanosine (8-oxo-dG) in peripheral lymphocyte DNA and 8-isoprostaglandin F-2alpha (8-iso-PGF2alpha) excreted in urine. Subjects consuming the high versus low VF diet had lower concentrations of 8-oxo-dG (p < 0.01) and of 8-iso-PGF2alpha (p < 0.01). However, the reduction in oxidative end points by high VF was not uniform. Rather, an antioxidant effect was observed primarily in individuals whose oxidative end points at baseline were above the median for the study population. Using change in plasma carotenoids (end point minus baseline measurements) as an index of phytochemical intake, the reduction in oxidative markers was inversely proportional to change in plasma carotenoids; this effect was stronger for lipid peroxidation (p < 0.01) than DNA oxidation (p < 0.05). These findings imply that increasing exogenous antioxidant exposure may primarily benefit individuals with elevated levels of oxidative stress. Null findings do not necessarily indicate that an antioxidant compound lacks in vivo activity.     

Land Disposal of Centrate from Biosolids Production

Arid land and riparian plants were irrigated with a liquid wastewater stream (centrate) from biosolids production at a municipal sewage treatment facility. Centrate containing approximately 1000 mg/l ammonia nitrogen was diluted with Tucson city tap water to provide treatments of 2.5, 5, 10, 25, 50, and 100% centrate, along with a dilution water control. Plants grown in 19-l containers in a greenhouse were irrigated daily with 6.7 l of solution over a 7-week period. All species exhibited a fertilizing effect at concentrations up to 5 and 10% centrate, equivalent to approximately 50 and 100 mg/l NH₃–N, but then inhibitory above 100 mg/l. No plants survived on the 50 or 100% treatments. If centrate were to be diverted from the wastewater facility to be disposed of on the land, it was estimated that the total nitrogen content of the biosolids produced by the plant would be reduced by 11%. This reduction would have only a small impact on the value of the biosolids for soil amendment applications.     

Quantification of seasonal sediment and phosphorus transport dynamics in an agricultural watershed using radiometric fingerprinting techniques

Purpose

Phosphorus (P) is a limiting nutrient for most US Midwestern aquatic systems and, therefore, increases of P, through point or non-point sources (NPS) of pollution such as agriculture, causes eutrophication. Identifying specific NPS contributions (e.g., upland vs. stream channels) for sediments and P is difficult due to the distributed nature of the pollution. Therefore, studies which link the spatial and temporal aspects of sediment and P transport in these systems can help better characterize the extent of NPS pollution.

Materials and methods

Our study used fingerprinting techniques to determine sources of sediments in an agricultural watershed (the North Fork of the Pheasant Branch watershed; 12.4 km² area) in Wisconsin, USA, during the spring, summer, and fall seasons of 2009. The primary sources considered were uplands (cultivated fields), stream bank, and streambed. The model used fallout radionuclides, ¹³⁷Cs, and ²¹⁰Pb_xs, along with total P to determine primary sediment sources. A shorter-lived fallout radioisotope, ⁷Be, was used to determine the sediment age and percent new sediments in streambed and suspended sediment samples (via the ⁷Be/²¹⁰Pb_xs ratio).

Results and discussion

Upland areas were the primary source of suspended sediments in the stream channels followed by stream banks. The sediment age and percent new sediment for the streambed and suspended sediments showed that the channel contained and transported newer (or more recently tagged with ⁷Be) sediments in the spring season (9–131 days sediment age), while relatively old sediments (165–318 days) were moving through the channel system during the fall season.

Conclusions

Upland areas are the major contributors to in-stream suspended sediments in this watershed. Sediment resuspension in stream channels could play an important role during the later part of the year. Best management practices should be targeted in the upland areas to reduce the export of sediments and sediment-bound P from agricultural watersheds.     

Optimization of chick embryotoxicity bioassay for testing toxicity potential of fungal metabolites

The optimization of a simple, sensitive procedure using a chick embryotoxicity screening test (CHEST) bioassay for detection of toxic compounds is presented. Dosing protocols of eggs, using several mycotoxins (aflatoxin B1, deoxynivalenol, T-2 toxin) and appropriate controls, were evaluated for embryonic sensitivity, overall practicality of the procedure, and consistency of results. It was found that both type of carrier solvent and volume injected could significantly affect overall embryonic mortality. The chick embryo was most sensitive to the effects of toxins and solvents after 1 or 2 days of incubation; a rapid decrease in response was observed as the age of the embryo at dosing increased. Following administration of the toxins just below the shell membrane by way of a small hole (less than 0.5 mm diameter) punched in the shell, a good dose-response (% mortality) could be obtained regardless of the site of injection (except directly into the yolk), although dosing via the air sac position resulted in a slightly better statistical outcome. Although some variations in calculated LD50 values were found among repeated assays, statistical analyses showed that the differences were not due to dosing protocol but to the variations in embryo sensitivities among batches of eggs. Thus, if standard reference toxins for comparison are run concurrently, the CHEST assay can prove to be a very satisfactory model, as well as having considerable flexibility to be adapted to the needs and resources of many laboratories.     

Paleontological Evidence of Varitions in Length of Synodic Month since Late Cambrian

The values of length of synodic month, obtained from tidally controlled periodical growth patterns in mollusks and stromatolites for several geologic periods, indicate that the deceleration rate of the earth's rotation has not been constant. Two breaks in slope, in the Pennsylvanian and Cretaceous, may be related to changes in distribution of continents, oceans, and adjacent shallow seas.     

Problems of official methods and new techniques for analysis of foods and feeds for vitamin A

The official methods for measuring vitamin A and carotene in foods and feeds have evolved over several decades. New procedures are needed to permit the use of modern analytical techniques such as liquid chromatography (LC), and to expand the application of official methods to a wide variety of foods. Recent improvements in methodology are reviewed. Emphasis is often placed on the separations achieved by LC, but all factors that affect quantitation, such as sampling, preparation of standards, and protection of the vitamin against oxidation, must be considered in the development of new methods. Many approaches to analysis are possible because of the versatility of LC but not all of them can be used in official methods. Methods for milk, margarine, and feeds can be prepared now for collaborative assays. Although straightforward methods can be developed for the measurement of beta-carotene in certain foods, a full examination of provitamins will probably continue to require a flexible approach and special expertise.     

Production and water use in lettuces under variable water supply

The effects of a variable water supply on the water use, growth and yield of two crisphead and one romaine (i.e., Cos) lettuce cultivar were examined in a field experiment using a line source sprinkler system that produced a range of water regimes that occur in growers fields. Four locations at increasing distances from the main line were monitored through the season (i.e., from thinning to harvest, 28–63 days after planting (DAP)). These locations at the end of the season corresponded to: (1) rewatering to field capacity (FC); (2) watering with a volume 13% below that required in the field capacity treatment (0.87*FC); (3) 30% below FC (0.70*FC); and (4) 55% below FC (0.45*FC). A linear production function for dry matter accumulation and fresh weight vs. crop evapotranspiration (ET_c) was determined for lettuce during this period, giving a water use efficiency for dry matter of 1.86 g m^–2 mm^–1 and for fresh weight of 48 g m^–2 mm^–1 . For lettuce irrigated to field capacity, ET_c between thinning and harvest was 146 mm; maximum crop coefficients of 0.81–1.02 were obtained at maturity (55–63 DAP). For the three irrigation treatments receiving the largest water application, ET_c was higher in the Cos culivar than in the two crisphead lettuce cultivars which had similar ET_c. Plant fresh weight was more sensitive than dry weight to reduction in water supply. In the FC treatment, root length density and soil water extraction were greatest in the top 0–45 cm, and decreased rapidly below 45 cm depth. Soil water extraction by roots increased at lower depths when irrigation was reduced. Instantaneous rates of leaf photosynthesis and leaf water potential showed no response to the irrigation treatments in this study, despite differences in biomass production. Evaporation was determined to be the major component of ET_c for 45 of the 63 days of the growing season. The large loss of water by evaporation during mid-season and the apparent insensitivity of lettuce to the volume of irrigation during this period may provide an opportunity for reducing irrigation applications.