Rapid enzyme immunoassays for the detection of carbaryl and methoprene in grains

In order for grain handlers and traders to reliably estimate residues of grain protectants in the field, antibody-based rapid tests were developed for carbaryl (1-naphthyl methylcarbamate) and methoprene [isopropyl (E,E)-(RS)-11-methoxy-3,7,11-trimethyldodeca-2,4-dienoate]. To complement the rapid analysis, a simple and rapid extraction technique was developed. In these tests, a pesticide-containing methanol extract of the grain sample and an enzyme-labeled component are added to precoated strips. After a brief incubation, the strips are washed and a substrate/chromogen for the enzyme is added. The color developed is stopped by acidification and the results are read either by eye or in a portable field photometer. The overall test time is under 20 minutes. For carbaryl, the test had a limit of detection of 4.5 ppb (1.1 ppm in grain), while the methoprene test had a limit of detection of 4 ppb (1 ppm in grain) based on the lower datum point, which is 15% inhibition, in the standard curves. Both assays can be used as a screening test for carbaryl and methoprene in animal feed grains.     

Distribution and persistence of carbaryl in some terrestrial and aquatic components of a forest environment

Abstract

An oil‐based formulation of carbaryl (1‐naphthyl N‐methyl‐carbamate) (Sevin‐2‐Oil) was applied twice by a fixed‐wing aircraft at a dosage rate of 280 g of A.I./ha/application to a coniferous forest near Allardville, New Brunswick. The highest concentrations of the chemical in fir foliage, litter and forest soil 1 h after application were respectively 4.20, 1.21 and 0.59 ppm (fresh weight). The residues dissipated rapidly and the DT50 values obtained from the depletion curves were 2.3 d for foliage and 1.5 d for litter and soil samples. Very low levels (<0.1 ppm) of carbaryl persisted in foliage and litter beyond the 10 d sampling period. The maximum residue level found in stream water was 0.314 ppm and more than 50% of it had dissipated within 1 h. Low but detectable levels (0.001 ppm) of the chemical persisted in water until the end of the 10 d sampling period. Sediment samples contained a maximum level of 0.04 ppm, which dissipated below the detection limit within 5 h. Brook trout and slimy sculpins captured in the stream 1 d after the spray contained on average about 0.04 ppm of carbaryl and none of it was found in 3 d postspray samples.     

Allium cepa derived EROD as a potential biomarker for the presence of certain pesticides in water

Allium cepa root length inhibition test is a well recommended bioassay for the evaluation of the toxicity of various polluted waters. The utility of EROD (7-ethoxy resorufin O-deethylase) as a potential biomarker of pesticide pollution was investigated using the Allium cepa system. Onion bulbs exposed to model water samples containing any of the six pesticides viz. 2,4-D, HCB, malathion, carbaryl, DDT and endosulphan were analyzed for EROD activity. The pesticide treatment resulted in the enhanced activity of the enzyme, with carbaryl and HCB causing 63- and 53-fold induction respectively with respect to the control at a dose of 1.2 ppb. The industrial wastewater samples from Ghaziabad city of Northern India resulted in about a 68-fold rise in the EROD activity, whereas the Aligarh samples did not exhibit any change within the statistical limit. These results suggest the presence of the test pesticides in the Ghaziabad sample and their absence in the Aligarh sample. Pesticide analysis in the test water samples by HPLC supported this to a large extent. Presence of cycloheximide in the test system brought down the EROD activity, equal to that of control, suggesting the de novo synthesis of the enzyme following the exposure of Allium cepa to pesticides. These studies suggest that the Allium cepa derived EROD can act as a potential biomarker of certain pesticides since even 1ppb of total/individual pesticides brought about >10-fold induction of EROD. We recommend the assay of EROD in the Allium cepa system as a presumptive test for the detection of these pesticides before using analytical techniques like HPLC.     

Validation of a test method for the measurement of methanol emissions from stationary sources

Title III of the 1990 Clean Air Act Amendments designated methanol as a pollutant to be regulated. The U.S. Environmental Protection Agency (EPA), through a contract with Research Triangle Institute, has developed a method for measuring methanol emissions from stationary sources. The methanol sampling train (MST) consists of a glass-lined heated probe, two condensate knockout traps, and three sorbent cartridges packed with Anasorb 747. Samples are desorbed with a 1:1 mixture of carbon disulfide (CS2) and N,N-dimethylformamide (DMF). Condensate water and CS2/ DMF samples are analyzed by gas chromatography with flame ionization detection. The MST has a practical quantitation limit of approximately 3 ppm for a 20-L sample. Samples were shown to be stable for at least two weeks after collection. Field tests of the MST and the National Council of the Paper Industry for Air and Stream Improvement (NCASI) methanol sampling method were conducted at two pulp and paper mills. Sampling and analysis procedures followed EPA Method 301 requirements. The sampling location for the first field test was the inlet vent to a softwood bleach plant scrubber, where the methanol concentration was approximately 30 ppm. The mean recovery of spike was 108.3% for the MST method and 81.6% for the NCASI method. Although neither method showed significant bias at the 95% confidence level, the between-methods bias was significantly different. A second field test was conducted at a vent from a black liquor oxidation tank where the methanol concentration was approximately 350 ppm. Mean spike recoveries were 96.6 and 94.2% for the MST and NCASI methods, respectively. The biases of the two methods and the between-methods bias were not significantly different for the second field test.     

Colloidal gold probe-based immunochromatographic assay for the rapid detection of lead ions in water samples

One-step immunochromatographic assay (ICA) has been developed using colloidal gold-labeled monoclonal antibody probe for the rapid detection of lead ions in water samples. The ICA was based on the theory of competitive reactivity, and the results can be easily judged based on the presence or absence of a red colored test line with visual detection. Under optimal conditions, this method shows high detecting sensitivity with a LOD (limit of detection) of 50 ng/ml. Stability test indicates that the immunochromatographic strips are stable for 8 weeks at room temperature. During practical application, nanometer TiO₂ is used to enrich the lead ions in water samples. The ICA is successfully applied in the measurement of lead ion concentrations in local water samples, and the results are highly consistent with that of ICP-MS. Detecting lead ions with ICA can be done within 4 min and is very useful for the rapid onsite testing.     

Cantilever nanobiosensor using tyrosinase to detect atrazine in liquid medium

The aim of this study was to develop a cantilever nanobiosensor for atrazine detection in liquid medium by immobilising the biological recognition element (tyrosinase vegetal extract) on its surface with self-assembled monolayers using gold, 16-mercaptohexadecanoic acid, 1-ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride/n-hydroxysuccinimide. Cantilever nanobiosensors presented a surface compression tension increase when atrazine concentrations were increased, with a limit of detection and limit of quantification of 7.754 ppb (parts per billion) and 22.792 ppb, respectively. From the voltage results obtained, the evaluation of atrazine contamination in river and drinking water were very close to those of the reference sample and ultrapure water, demonstrating the ability of the cantilever nanobiosensor to distinguish different water samples and different concentrations of atrazine. Cantilever nanosensor surface functionalization was characterised by combining polarisation modulation infrared reflection-absorption spectroscopy and atomic force microscopy and indicating film thickness in nanometric scale (80.2 ± 0.4 nm). Thus, the cantilever nanobiosensor developed for this study using low cost tyrosinase vegetal extract was adequate for atrazine detection, a potential tool in the environmental field.     

A method for the determination of rhodamine b and brilliant sulfaflavine on cotton string collectors and in a spray tank solution mixture

Abstract

A fluorescent method for the determination of rhodamine B (RhB) and brilliant sulfaflavine (BSF) dual tracers on cotton string collectors and in spray tank solutions was developed for the evaluation of the effects of adjuvants on the atmospheric drift of agricultural sprays. Both tracers on collectors were extracted with deionized water simultaneously and measured directly with a Fluorescent Specrometer by switching the maximum excitation and emission wavelengths. The linear ranges of the RhB and BSF standard curves were 0–50 and 0–10 ppb respectively. Recoveries of tracers on cotton strings ranged from 92.9% ± 1.1% to 95.1% ± 1.0% for RhB and 91.9% ± 2.8% to 103% ± 0.6% for BSF when spiked levels ranged from 0.25 to 2.5 μg per collector. The detection limit of the instrument was approximately 0.1 ppb for both tracers. The detection limits of the analytical method were approximately 0.1 ppb for RhB and 0.3 ppb for BSF when the background levels of cotton strings were subtracted. The presence of one tracer in the solution did not significantly interfere with the detection of the other tracer in the same solution. The presence of adjuvants added to the spray tank solutions did not interfere with the detection of both tracers when the adjuvants were applied at the manufacturer's maximum recommendation rates.     

Free Enterprise and Air Pollution Control

Abstract

Title III of the 1990 Clean Air Act Amendments designated methanol as a pollutant to be regulated. The U.S. Environmental Protection Agency (EPA), through a contract with Research Triangle Institute, has developed a method for measuring methanol emissions from stationary sources. The methanol sampling train (MST) consists of a glass-lined heated probe, two condensate knockout traps, and three sorbent cartridges packed with Anasorb® 747. Samples are desorbed with a 1:1 mixture of carbon disulfide (CS₂) and N, N-dimethylformamide (DMF). Condensate water and CS₂/ DMF samples are analyzed by gas chromatography with flame ionization detection. The MST has a practical quantitation limit of approximately 3 ppm for a 20-L sample. Samples were shown to be stable for at least two weeks after collection.

Field tests of the MST and the National Council of the Paper Industry for Air and Stream Improvement (NCASI) methanol sampling method were conducted at two pulp and paper mills. Sampling and analysis procedures followed EPA Method 301 requirements.

The sampling location for the first field test was the inlet vent to a softwood bleach plant scrubber, where the methanol concentration was approximately 30 ppm. The mean recovery of spike was 108.3% for the MST method and 81.6% for the NCASI method. Although neither method showed significant bias at the 95% confidence level, the betweenmethods bias was significantly different.

A second field test was conducted at a vent from a black liquor oxidation tank where the methanol concentration was approximately 350 ppm. Mean spike recoveries were 96.6 and 94.2% for the MST and NCASI methods, respectively. The biases of the two methods and the between-methods bias were not significantly different for the second field test.     

A rapid and sensitive method for methyl tert-butyl ether analysis in water samples by use of solid phase microextraction and gas chromatography-mass spectrometry

This work describes a rapid and sensitive solid-phase microextraction (SPME) method for the isolation and analysis of methyl tert-butyl ether in water samples. Methyl tert-butyl ether was extracted from aqueous solutions using SPME fibre coated with Divinylbenzene/Carboxen/polydimethylsiloxane (30 microm film thickness) and analysed by GC-MS with a Hewlett Packard 6890/5973 system equipped with a capillary column coated with Vocol (30 m x 0.25 mm, 1.5 microm film thickness). Extraction parameters and chromatographic separation conditions were optimised. The developed method showed good analytical performance in terms of precision (RSD between 2% and 8%) and accuracy (mean recovery from 96% to 104%) with a detection limit of 14 ppt. Finally the method was applied to surface, tap and commercial mineral water samples, as well as snow samples collected along a busy road of Bologna town area. The median concentration of methyl tert-butyl ether in all these samples (0.05-0.4 ppb) was well below the maximum aqueous contamination levels in water adopted in the United States (13 ppb).     

Disruption of hepatic mitochondrial bioenergetics is not a primary mechanism for the toxicity of methoprene - relevance for toxicological assessment

Methoprene (isopropyl(2E,4E)-11-methoxy-3,7,11-trimethyl-2,4-dodecadienoate) is an insect growth regulator generally used to control insect populations by preventing insect maturation. So far, the effects of the insecticide on mitochondrial bioenergetics were not investigated. In the present work, liver mitochondria from Wistar rats were isolated and features of mitochondrial physiology were characterized in the presence of methoprene. High concentrations of methoprene, in the range of 40-100 nmol/mg of protein could decrease the transmembrane electric potential (Delta Psi) developed by mitochondria and, at the highest concentration, methoprene prevented complete Delta Psi repolarization after ADP addition. The effect was more evident using succinate than with ascorbate+TMPD as substrate. State 3 respiration was approximately 60% inhibited by 80 nmol of methoprene/mg of protein, while state 4 respiration, within the same range of methoprene concentrations, showed a slight increase, when both glutamate-malate and succinate were used as substrates. Additionally, FCCP-stimulated respiration was inhibited to an extent comparable to the effect on state 3, which suggests an interaction of methoprene with the respiratory chain, more evident with glutamate/malate as substrate. The activity of complex I (NADH-ubiquinone oxidorreductase) and that of the segment comprehending complexes II and III (succinate-cytochrome c reductase) were decreased in the presence of methoprene (approximately 60% and 85% of inhibition, respectively, with 300 nmol of methoprene/mg of protein), while the activities of cytochrome c oxidase and ATPase do not seem to be affected. Furthermore, the action of methoprene on the mitochondrial permeability transition was also studied, showing that the insecticide (in the range of 30-80 nmol mg(-1) of protein) decreases the susceptibility of liver mitochondria to the opening of the transition pore, even in non-energized mitochondria. These results lead to the conclusion that methoprene interference with hepatic mitochondrial function occurs only for high concentrations, which implies that the noxious effects of the insecticide reported for a number of non-target organisms are not fully attributable to mitochondrial effects. Therefore, it seems that mitochondrial activity does not represent the primary target for methoprene toxic action.     

Improved solid-phase enzyme immunoassay systems in the ppt range for atrazin in fresh water

A solid-phase enzyme immunoassay (EIA) in poylstyrene microtiter-plates was developed for the rapid determination of atrazin in fresh waters. The linear measuring range was between 1 and 2000 p mol atrazin per assay. Atrazin containing aqueous solutions could be directly assayed in a concentration range of 1.1 up to 2200 μg/1 (ppb). The sensitivity of this EIA variant was improved in two ways. Using polystyrene spheres as antibody carriers, a detection limit of 11 ng/1 (ppt) could be reached. If an affinity purified antiserum was applied, detection limits of 11 ng/1 (ppt) in case of microtiter-plates and 0.1 ng/1 (ppt) in case of polystyrene spheres as antibody carriers were obtained. The cross reactivity of other s-triazines in the atrazin EIA was tested.     

Detection Limits for Hazardous and Radioactive Elements in Airborne Aerosols Using Inductively Coupled Air Plasma – Atomic Emission Spectrometry

Abstract

This research presents investigations into the use of inductively coupled air plasma – atomic emission spectrometry (air-plasma ICPAES) to determine the presence of inorganic contaminants in airborne aerosols. Limits of detection (LOD) in the ppm to ppb range for 19 hazardous metals and radionuclides were determined for aerosols of solutions nebulized into the air plasma. For many elements, the determined LOD surpass the threshold limit values established by the American Conference of Governmental Industrial Hygienists by one to three orders of magnitude. The potential of air-plasma ICPAES for continuous on-line monitoring of airborne contaminants is discussed.     

High-performance liquid chromatography with conductimetric detection of perfluorocarboxylic acids and perfluorosulfonates

A rapid and simple method for separating and determining various environmentally harmful perfluorocarboxylic acids and perfluorosulfonates was successfully developed using high- performance liquid chromatography with conductimetric detection, for product and waste management of these compounds at manufacturing and processing sites. Compounds having C(3)-C(8) perfluoroalkyl groups were separated using a Tosoh TSKgel Super-ODS column and a mobile phase consisting of a mixture of methanol and aqueous NaH(2)PO(4) at several mixing ratios. The best detection limits for the compounds ranged from 0.12 to 0.66 mg l(-1) (ppm), and linear calibration graphs were obtained up to 87-109 mg l(-1). The combination of this method with concentration of the sample by solid-phase extraction with cartridges based on styrene-divinylbenzene-copolymer enabled the determination of approximately 50 microg l(-1) (ppb) for compounds with C(4)-C(8) perfluoroalkyl groups. This method was successfully used to monitor the artificial decomposition of the perfluorocarboxylic acid n-C(4)F(9)COOH induced by a photocatalyst.     

The dissipation of nonlyphenol in stream and pond water under simulated field conditions

Abstract

The dissipation of 1.0 ppm nonylphenol in stream and pond water, incubated in flasks at 16°C under simulated field conditions up to 44 days indicated that the half‐life was 2.5 days if the flasks were open, and 16 days if they were closed. A transformed product was detected in the closed flasks.

Translocation of nonylphenol in water occurred when treated water samples were incubated in the presence of sediment. After 10 days, nonylphenol was detected only in the sediment, but not in water (detection limit = 10 ppb). About 80% of the nonylphenol was degraded in 71 days, but no degradation occurred if the water and the sediment were autoclaved prior to incubation.     

Tube-immunoassay for rapid detection of carbaryl residues in agricultural products

An antibody-based rapid, quantitative, and qualitative tube enzyme-linked immunosorbent assay (tube-ELISA) was developed and used to determine carbaryl (1-naphthyl methylcarbamate) residues in agricultural products (apple, Chinese cabbage, rice, and barley). The tube-ELISA is a competitive immunoassay in which the antibody is coated in the polystyrene tube, with a dynamic range between 0.7 and 46.3 microg kg(-1). Carbaryl was extracted from each agricultural sample by hand-shaking with methanol and examined for application to on-site analysis. After the liquid extraction, the sample extracts diluted with buffer were analyzed by rapid tube-ELISA directly. The overall test time was around 15-30 min, including sample preparation and assay performance. The results obtained from tube-ELISA correlated well with high-performance liquid chromatography (R2 > 0.9). The study shows that tube-ELISA is useful as a quality control tool and can be used to quantitatively detect carbaryl as well.     

Laboratory and field evaluation of measurement methods for one-hour exposures to O3, PM2.5, and CO.

While researchers have linked acute (less than 12-hr) ambient O3, PM2.5, and CO concentrations to a variety of adverse health effects, few studies have characterized short-term exposures to these air pollutants, in part due to the lack of sensitive, accurate, and precise sampling technologies. In this paper, we present results from the laboratory and field evaluation of several new (or modified) samplers used in the "roll-around" system (RAS), which was developed to measure 1-hr O3, PM2.5, and CO exposures simultaneously. All the field evaluation data were collected during two sampling seasons: the summer of 1998 and the winter of 1999. To measure 1-hr O3 exposures, a new active O3 sampler was developed that uses two nitrite-coated filters to measure O3 concentrations. Laboratory chamber tests found that the active O3 sampler performed extremely well, with a collection efficiency of 0.96 that did not vary with temperature or relative humidity (RH). In field collocation comparisons with a reference UV photometric monitor, the active O3 sampler had an effective collection efficiency ranging between 0.92 and 0.96 and a precision for 1-hr measurements ranging between 4 and 6 parts per billion (ppb). The limits of detection (LOD) of this method were 9 ppb-hr for the chamber tests and approximtely 16 ppb-hr for the field comparison tests. PM2.5 and CO concentrations were measured using modified continuous monitors--the DustTrak and the Langan, respectively. A size-selective inlet and a Nafion dryer were placed upstream of the DustTrak inlet to remove particles with aerodynamic diameters greater than 2.5 microm and to dry particles prior to the measurements, respectively. During the field validation tests, the DustTrak consistently reported higher PM2.5 concentrations than those obtained by the collocated 12-hr PM2.5 PEM samples, by approximately a factor of 2. After the DustTrak response was corrected (correction factor of 2.07 in the summer and 2.02 in the winter), measurements obtained using these methods agreed well with R2 values of 0.87 in the summer and 0.81 in the winter. The results showed that the DustTrak can be used along with integrated measurements to measure the temporal and spatial variation in PM2.5 exposures. Finally, during the field validation tests, CO concentrations measured using the Langan were strongly correlated with those obtained using the reference method when the CO levels were above the LOD of the instrument [approximately 1 part per million (ppm)].     

Comparative effects of mine wastes on the benthos of an acid and an alkaline pond

Benthic invertebrates collected from an acid (pH 3.2) and an alkaline (pH 7.9) tailing pond in the Rouyn-Noranda mining region were compared with those of a control pond in order to assess the effects of the pH on the benthic community structure and on metal accumulation by invertebrates. The acid pond showed high mean soluble concentrations of aluminium (2600 ppb), copper (230 ppb), iron (6500 ppb) and zinc (4900 ppb) in the water and high mean concentrations of copper (340 ppm), iron (15,238 ppm) and zinc (445 ppm) in the sediments. The benthic fauna was dominated by Chironomus spp and the water beetle Ilybius sp. The Shannon-Weaver diversity index was 0.04. The alkaline pond had high mean sediment concentrations of cadmium (21 ppm), iron (15,823 ppm), manganese (658 ppm) and nickel (146 ppm). A total of 37 insect genera and a diversity index of 3.4 were recorded for this pond. Of the seven metals studied only copper and aluminium showed greater accumulations in the biota of the contaminated ponds than in that of the control. Insects dominated the benthic fauna in both contaminated ponds and crustacea in the control.     

Effects of five insecticides used in apple orchards on Hyaliodes vitripennis (Say) (Hemiptera: Miridae)

Azinphos-methyl, carbaryl, dimethoate, phosmet and phosalone were used in apple orchards to manage apple aphid, apple maggot, woolly apple aphid and leaf eating caterpillars. Among the five insecticides evaluated, dimethoate, carbaryl and azinphosmethyl were the most toxic to the nymphs and adults of Hyaliodes vitripennis (Say) from two regions. Phosalone was the least toxic. Nymphs were more resistant than the adults. While the LC50 for dimethoate was 130 ppm for nymphs, it was 3 ppm for adults from St. Jean-Baptiste-de-Rouville. There were also significant differences in the level of resistance between the two regions where the H. vitripennis were collected. At St. Alexandre the LC50 for phosalone on nymphs was 19,250 ppm whereas, at St. Jean-Baptiste-de-Rouville it was 160,000 ppm.     

Determination of ozone in outdoor and indoor environments using nitrite-impregnated passive samplers followed by ion chromatography

An improved ion chromatographic (IC) method has been developed for the separation of nitrate in filter extracts in the presence of high concentrations of nitrite. This analytical method was successfully used for an indirect measurement of ozone (O3) in outdoor and indoor air, following its collection using a nitrite-impregnated passive sampler. The limit of detection and the limit of quantification, using the modified IC method, were 6 microg l(-1) (3sigma) and 20 microg l(-1) (10sigma), respectively. Improved detection limits and low baseline noise were obtained with the use of eluent generator and high-capacity ion exchange column. The optimized method was used for assessing O3 concentration in both indoor and outdoor environments of 28 child care centers (CCCs) located in different parts of Singapore. The O3 concentrations ranged from 0.1 to 11.95 parts per billion (ppb) in indoor and from 3.2 to 21.7 ppb in outdoor environments during the study period. It was found that, among the CCCs investigated in this study, air-conditioned CCCs and those located in close proximity to traffic emissions had significantly lower O3 concentrations indoors.     

Persistence characteristics of operationally sprayed fenitrothion in nearby unsprayed areas of a conifer forest ecosystem in New Brunswick

Abstract

Persistence characteristics of operationally sprayed fenitrothion were investigated in various substrates sampled from neighbouring unsprayed areas in New Brunswick. Air, water, sediment, aquatic plants, fish, balsam fir [Abies balsamea (L.) Mill] foliage, forest soil and litter samples were collected from random sampling locations selected within 200 m from the operational spray blocks. The same substrates were resampled from the same plots and from the same locations about a year later just prior to the commencement of the operational spraying. Control samples were collected from an unsprayed site, near Sault Ste. Marie, Ontario. All samples were analysed for fenitrothion, by gas‐liquid chromatography. Except the fish samples all the substrates collected during the time of operational spraying contained low but detectable levels of fenitrothion. When collected a year later prior to the operational spray program, only balsam fir showed any detectable levels (detection limit, 0.01 ppm) of the chemical. All other samples showed no fenitrothion residues (detection limit for air, 10 ng/m³; for water, 0.01 ppb; and for other samples, 0.01 ppm). The findings confirmed that fenitrothion does not persist for an extended period of time in the aquatic substrates. The conifer foliage, however, showed persistent residues at a level of about 0.55 ppm even after the winter months, although there was no indication of accumulation of the chemical as a result of repeated exposure. The study demonstrated that the conifer needles acted as a micro sink for the chemical which showed a tendency to persist in the leaf tissues for a considerable length of time.