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PubMed2002

Comparison of microextraction procedures to determine pesticides in oranges by liquid chromatography-mass spectrometry.

A liquid chromatographic-mass spectrometric method has been developed for the determination of bitertanol, carbendazim, fenthion, flusilazole, hexythiazox, imidacloprid, methidathion, methiocarb, pyriproxyfen and trichlorfon. Two procedures, based on stir bar sorptive extraction (SBSE) and matrix solid-phase dispersion (MSPD), have been evaluated for the extraction of these compounds in oranges. Their respective advantages and disadvantages are also discussed. The recoveries obtained by MSPD ranged from 47 to 96% and the relative standard deviations (RSDs) ranged from 1 to 15%, whereas with the SBSE method the recoveries were between 8 and 84% and the RSDs between 4 and 16%. Although, the limits of quantitation of most compounds are much better (0.001-0.05 mg kg(-1)) by SBSE, it is not suitable to determine some polar pesticides as carbendazim, imidacloprid and trichlorfon. Results obtained by both methods were compared, in terms of sensitivity and selectivity, with a classical ethyl acetate extraction method, and the three methods were applied to analyze real samples. As MSPD is easier to perform, faster than the organic solvent extraction, and shows equal accuracy and resolution, its application for analyzing pesticides in oranges is recommended.

Chromatography, Liquid↗
PubMed2003

Fast supercritical fluid extraction and high-resolution gas chromatography with electron-capture and flame photometric detection for multiresidue screening of organochlorine and organophosphorus pesticides in Brazil's medicinal plants.

A description is given of a rapid and environmentally friendly method to determine organochlorine and organophosphorus pesticide multiresidues--malathion, methidathion, fenitrothion, fenthion, parathion-ethyl, parathion-methyl, lindane, hexachlorobenzene, chlorothalonil, tetradifon, alpha-endosulfan, beta-endosulfan and dieldrin-in Passiflora alata Dryander and pasiflora edulis Sims. f. flavicarpa Deg. leaves by supercritical fluid extraction and high-resolution gas chromatography with electron-capture and flame photometric detection (HRGC-ECD/FPD). The mild extraction conditions [pure CO2; 100 bar (1 bar = 10(5) Pa) and 40 degrees C (p = 0.62 g/ml); 5 min static+10 min dynamic extraction time; ODS trap and elution with 1 ml n-hexane at 2 ml/min] allow for direct analysis by HRGC-ECD/FPD with no prior cleaning procedure. The method provides. in accordance with the validation criteria of the European Pharmacopoeia, analytical results that are similar or even better than the official procedures, and is simpler, faster and cheaper. Mean recoveries of 69.8-107.1% were obtained, with 1.4-14.7% reproducibility (RSD). The method was applied to analyse commercial samples of Passiflora L. from Brazil. Twenty-three percent of the samples showed the presence of the organochlorine or organophosphorus pesticide residue investigated.

Chromatography, Gas↗
PubMed2003

Headspace solid-phase microextraction in combination with gas chromatography-mass spectrometry for the rapid screening of organophosphorus insecticide residues in strawberries and cherries.

A headspace solid-phase microextraction (HS-SPME) method in combination with GC-MS was used for the extraction and quantification of diazinon, fenitrothion, fenthion, parathion ethyl, bromophos methyl, bromophos ethyl and ethion. The method was developed using a 100-microm poly(dimethylsiloxane) fiber. The obtained results showed higher responses of the insecticides after addition of aliquots of water and solvent to the fruit samples. Calibration curves that were constructed for the analytes spiked into strawberry and cherry samples followed linear relationships with good correlation coefficients (R2 > 0.986). Linearity range was between 50 and 500 microg/kg and the precision was found to be lower than 15% when applying the optimized HS-SPME procedure to fruit samples. Limits of detection in both strawberry and cherry samples using GC-MS (selected ion monitoring mode) were below 13 microg/kg. Moreover, the HS-SPME method was applied to the analysis of fruit samples and compared with liquid-liquid extraction. Results obtained in this study were in good agreement with those obtained using liquid-liquid extraction demonstrating that the recommended procedure was a fast, accurate and stable sample pretreatment method obtaining good efficiency for the extraction of organophosporus insecticides from strawberries and cherries.

Fragaria↗
PubMed1999

Validation of a solid-phase microextraction method for the determination of organophosphorus pesticides in fruits and fruit juice.

A method for the determination of organophosphorus pesticides (diazinon, fenitrothion, fenthion, quinalphos, triazophos, phosalon and pyrazophos) in fruit (pears) and fruit juice samples was developed and validated. The samples were diluted with water, extracted by solid-phase microextraction (SPME) and analysed by gas chromatography (GC) using a flame photometric detector in phosphorous mode. Limits of detection of the method for fruit and fruit juice matrices were below 2 micrograms/kg for all pesticides. Relative standard deviations for triplicate analyses of samples fortified at 25 micrograms/kg of each pesticide were not higher than 8.7%. Recovery tests were performed for concentrations between 25 and 250 micrograms/kg. Mean recoveries for each pesticide were all above 75.9% and below 102.6% for juice, and between 70 and 99% for fruit except for pyrazophos in the fruit sample (with mean recovery of 53%). Therefore, the proposed method is applicable in the analysis of pesticides in fruit matrices and the use of the method in routine analysis of pesticide residues is discussed.

Artifacts↗
PubMed2002

Removal of organophosphate pesticides from wastewater by supercritical carbon dioxide extraction.

Organophosphate pesticides including fenitrothion chlorpyrifos, diazinon, methamidophos, edifenphos, mevinphos, fenthion, and acephate present in agro-wastewater can be effectively removed by supercritical carbon dioxide (SC-CO2) extraction. Near quantitative removal of the pesticides from the aqueous solution can be achieved by SC-CO2 at 90 degrees C and 325 atm for 20 min of static extraction followed by 40 min of dynamic extraction. The extracted pesticides were collected in a small amount of Fenton's solution. The pesticides in Fenton's solution were degraded completely within an hour after the collection. A combination of SC-CO2 extraction and subsequent degradation by Fenton's reagent may provide an alternative water purification strategy for treating organophosphate pesticides in agro-wastewater.

Agriculture↗
PubMed1996

Tomato Prf is a member of the leucine-rich repeat class of plant disease resistance genes and lies embedded within the Pto kinase gene cluster.

In tomato, resistance to Pseudomonas syringae pv. tomato (Pst) strains expressing the avirulence gene avrPto requires the presence of at least two host genes, designated Pto and Prf. Here we report that Prf encodes a protein with leucine-zipper, nucleotide-binding, and leucine-rich repeat motifs, as are found in a number of resistance gene products from other plants. prf mutant alleles (4) were found to carry alterations within the Prf coding sequence. A genomic fragment containing Prf complemented a prf mutant tomato line both for resistance to Pst strains expressing avrPto and for sensitivity to the insecticide Fenthion. Prf resides in the middle of the Pto gene cluster, 24 kb from the Pto gene and 500 bp from the Fen gene.

Alleles↗
PubMed2001

Horn fly (Diptera: Muscidae) resistance to organophosphate insecticides.

Insecticidal ear tags impregnated with organophosphate (OP) insecticides were used each year from 1989 to 1998 at Rosepine, LA. Weekly fly counts were conducted to evaluate control efficacy of the treatments, and bioassays were conducted at least twice per year to measure fly susceptibility to OP and pyrethroid insecticides. Between 1989 and 1992, the efficacy of 20% diazinon-impregnated ear tags was reduced from >20 to just 1 week of control. A high risk of control failure was observed when a resistance frequency of approximately 5% was measured in pre-season bioassays. Resistance to diazinon, fenthion, ethion, pirimiphos-methyl, and tetrachlorvinphos was observed. Esterase activity toward alpha-naphthyl acetate was significantly higher in flies collected at Rosepine in 1997 than in flies from a laboratory colony and from a susceptible field population.

Administration, Topical↗
PubMed1996

Determination of parts per trillion levels of organophosphorus pesticides in groundwater by automated on-line liquid-solid extraction followed by liquid chromatography/atmospheric pressure chemical ionization mass spectrometry using positive and negative ion modes of operation.

Liquid chromatography/atmospheric pressure chemical ionization mass spectrometry with positive and negative ion modes of operations was used for the trace determination of several organophosphorus pesticides, (E)- and (Z)-mevinphos, dichlorvos, azinphos-methyl, azinphosethyl, parathion-methyl, parathion-ethyl, malathion, fenitrothion, fenthion, chlorfenvinphos, and diazinon, in groundwater. This method required only 100 mL of water, and it was combined with a prior automated online liquid-solid extraction step using an OSP-2 autosampler containing C18 cartridges. The limit of quantitation (LOQ) varied between 5 and 37 ng/L in positive ion (PI) mode. Under negative ion (NI) mode of operation, only the parathion group (both parathions and fenitrothion) had a better sensitivity as compared to that in PI mode, with a LOQ of 5-15 ng/L, whereas the rest of pesticides had 2-4 times higher LOQs as compared to those in PI mode. Selected ion monitoring of the group-specific fragment of the organophosphorus pesticides, e.g., [(CH3O)2PO2]- or the [M + H]+ ions, under NI or PI mode, respectively, was used. Sample cone voltage varied from 10 to 130 V. This parameter influenced the transmission and fragmentation of quasi-molecular ions, and it was optimized to achieve identification capabilities with the highest sensitivity. At 20 V, good fragmentation was obtained for most of the studied analytes. The system was used for the certification of a groundwater sample spiked at the nanograms per liter level with organophosphorus pesticides provided by Aquachek.

Chromatography, Liquid↗
PubMed2001

Use of cholinesterase activity in monitoring organophosphate pesticide exposure of cattle produced in tropical areas.

The use of cholinesterase activity as a biochemical method for monitoring organophosphate pesticide exposure in cattle is described herein. Determination of cholinesterase activity of whole blood, erythrocyte, and plasma was carried out according to the Ellman modified kinetic method. The mean baseline acetylcholinesterase activities of 9.549 +/- 3.619 IU/mL in whole blood, 9.444 +/- 3.006 IU/mL in erythrocytes, and 0.149 +/- 0.063 IU/mL in plasma were estimated for steers from the control group. Results of multivariate analysis showed that the general responses between the control and experimental groups (in vivo, monitoring and case studies) treated with Coumaphos and Fenthion were statistically different, and the general responses of these experimental groups were statistically different over time as well. Among the fractions that were analyzed, the erythrocyte acetylcholinesterase activity could be adequate for the diagnosis of exposure or acute poisoning in cattle as it showed a good within-run and between-run precision with CVs <10% better than those in plasma.

Acetylcholinesterase↗
PubMed1998

Determination of Organophosphorus Pesticide Residues in Greek Virgin Olive Oil by Capillary Gas Chromatography.

In this study, the occurrence of 15 organophosphorus pesticide residues in Greek virgin olive oil was investigated. Analysis was carried out using capillary gas chromatography with specific detectors (FPD and NPD), after sample extraction with n-hexane and cleanup by partitioning between n-hexane and acetonitrile. Sixty-two samples of virgin olive oil were taken from the major production areas and packing companies of Greece during 1992-1994. In 46 samples, 9 organophosphorus pesticides, namely dimethoate, fenthion, omethoate, chlorpyrifos, methamidophos, parathion-methyl, parathion, methidathion, and malathion, were found, in concentrations ranging from 0.0005 to 0.1800 mg/kg. Diazinon, pirimiphos-methyl, paraoxon-methyl, malaoxon, carbophenothion, and azinphos-ethyl were not detected in any sample. In most samples pesticide residues were below the detection limits (0.0001 and 0.001 mg/kg), and most of the positive findings were a fraction (i.e., <0.09-18%) of the FAO/WHO Codex Alimentarius maximum residue limits (MRLs) except for dimethoate, which was ranged between 1 and 45%. Only one sample contained dimethoate residue that exceeded the Codex MRL for refined olive oil.

Journal Article↗
PubMed2002

Solid-phase microextraction coupled to gas chromatography-mass spectrometry for the study of soil adsorption coefficients of organophosphorus pesticides.

A solid-phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS) method for the simultaneous determination of the organophosphorus pesticides (OPPs), phorate, diazinon, methyl-parathion, fenitrothion, malathion, fenthion, ethyl-parathion and methidathion, has been developed to study their soil/water distribution. The method was used in conjunction with a conventional 'batch equilibrium method' to assess the soil adsorption coefficients (Koc) of the target compounds in different soil samples with known organic carbon content. Contrary to traditional techniques, the present method is fast, solvent-free and highly sensitive, thus permitting the assessment of the Koc values of the target compounds even at low soil concentration levels, close to those encountered in real field contamination, where the Freudlich adsorption isotherms can be considered to be linear. The estimated Koc values were found to be in good agreement with those reported in the literature.

Chemistry Techniques, Analytical↗
PubMed1999

Comparative effectiveness of organophosphorus protoxicant activating systems in neuroblastoma cells and brain homogenates.

The ability of bromine and rat liver microsomes (RLM) to convert organophosphorus (OP) protoxicants to esterase inhibitors was determined by measuring acetylcholinesterase (AChE) and neuropathy target esterase (NTE) inhibition. Species specific differences in susceptibility to esterase inhibition were determined by comparing the extent of esterase inhibition observed in human neuroblastoma cells and hen, bovine, and rodent brain homogenates. OP protoxicants examined included tri-o-tolyl phosphate (TOTP), O-ethyl O-p-nitrophenyl phenylphosphonothioate (EPN), leptophos, fenitrothion, fenthion, and malathion. Bromine activation resulted in greater AChE inhibition than that produced by RLM activation for equivalent concentrations of fenitrothion, malathion, and EPN. For EPN and leptophos, bromine activation resulted in greater inhibition of NTE than RLM. Only preincubation with RLM activated TOTP; resultant inhibition of AChE was less in hen brain (13 +/- 3%) than in neuroblastoma cells (73 +/- 1%) at 10(-6) M. In contrast, 10(-6) M RLM-activated TOTP produced more inhibition of hen brain NTE (89 +/- 6%) than NTE of human neuroblastoma cells (72 +/- 7%). Human neuroblastoma cells and brain homogenates from hens, the accepted animal model for study of OP-induced neurotoxicity, were relatively similar in sensitivity to esterase inhibition. Homogenates from hens were more sensitive to NTE inhibition induced by phenyl saligenin phosphate (PSP), an active congener of TOTP, than were homogenates from less susceptible species (mouse, rat, bovine). AChE of hen brain homogenates was also more sensitive than homogenates from other species to malaoxon, the active form of malathion.

Animals↗
PubMed1983

Further water solubility determinations of insecticidal compounds.

The aqueous solubility of 39 insecticidal and related compounds was determined at 20 +/- 1.5 degrees C, using a previously described shaking and centrifugation method. Fenamiphos, fenthion and methidathion produced values substantially less than those reported in the literature whereas, aminocarb, diazinon, dicapthon, pirimiphos-ethyl and pirimiphos-methyl gave solubilities substantially greater than reported literature values.

Carbamates↗
PubMed2002

Simple determination of 22 organophosphorous pesticides in human blood using headspace solid-phase microextraction and gas chromatography with mass spectrometric detection.

A simple and rapid procedure for the determination of 22 organophosphorous pesticides (bromophos-ethyl, bromophos-methyl, chlorfenvinphos, chlorpyriphos, demethon-S-methylsulfon, diazinon, dichlorvos, dicrotophos, dimethoate, disulfoton, edifenphos, fenitrothion, fenthion, malathion, methidathion, mevinphos, monocrotophos, omethoate, parathion-ethyl, parathion-methyl, phosphamidon, and quinalphos) in human blood using headspace (HS) solid-phase microextraction (SPME) and gas chromatography (GC)-mass spectrometry (MS) is presented. The effects of various sample additions, incubation temperatures, absorption times, desorption times, and depths of fiber insertion into the injection port of the GC are optimized to enhance the sensitivity of the procedure. The recoveries of spiked blood samples are determined between 70% and 95% compared with samples prepared in water, and absolute recoveries are in the range between 0.1% and 19.6%. For quantitation in the single ion monitoring mode, linearity is established over concentration ranges from 0.025 to 5.0 microg/g with excellent coefficients of correlation (0.991-0.998). The detection limits are in the range between 0.01 and 0.3 microg/g. The time for analysis is 44 min per sample including extraction and GC-MS analysis. HS-SPME in combination with GC-MS is an effective method for the determination of organophosphorous pesticides in human blood and shows a great potential for use in rapid on-site analytical work, which is highly demanded in clinical and forensic toxicology.

Calibration↗
PubMed1989

Improved filter paper test for detecting and quantifying increased esterase activity in organophosphate-resistant mosquitoes (Diptera: Culicidae).

A previously described filter paper test procedure for detecting of esterases involved in organophosphate insecticide resistance in the Culex pipiens L. complex was modified to permit quantification of esterase activity and resistance in single insects. The new procedure, FP/Est test, was used to survey organophosphate resistance in 11 field collections from seven states. Clear discrimination of increased activity was possible by visual inspection and by densitometric analysis. The proportion of insects with susceptible-like esterase activity was strongly correlated with (and often was not significantly different from) the proportion found to be susceptible by bioassay with chlorpyrifos, temephos, fenthion, and malathion, indicating that the FP/Est test is a reliable method for detecting and monitoring of organophosphate resistance. In addition, the 90th percentile of esterase activity in each collection was significantly correlated with the LC90 of each of the four insecticides, suggesting that the FP/Est test also can be used as a rough estimate of resistance levels. Application of the FP/Est test to monitor resistance caused by increased esterase activity in mosquitoes and agricultural pests is discussed.

Animals↗
PubMed1997

Cytochrome P-450 field insecticide tolerance and development of laboratory resistance in grape vine populations of Drosophila melanogaster (Diptera: Drosophilidae).

Studies were conducted between 1993 and 1996 using 3 natural grape vine populations, 1 susceptible laboratory strain, and 1 resistant selected strain of Drosophila melanogaster L. In vitro monooxygenase activity (ethoxycoumarine-O-deethylation) (ECOD) was recorded from microsomal fractions of all strains. Results varied over a 6-fold range between susceptible laboratory Canton and resistant selected RDDT strains and over a 2-fold range between the Canton strain and natural populations of flies. Few significant variations of ECOD activity were detected among the natural populations despite many insecticide treatments, but activities were significantly correlated with toxicological tolerance to 5 of the 15 insecticides (deltamethrin, fipronil, chlorpyriphos ethyl, DDT, and diazinon). Moreover, immunoblotting responses of microsomal protein encoded by Cyp6A2 showed that the levels of expression were quantitatively correlated with toxicological tolerance to almost the same group of insecticides (deltamethrin, fipronil, chlorpyriphos ethyl, DDT, fenvalerate, and fenthion). However, the level of CYP6A2 expression in some natural strains (still weakly resistant) was almost comparable with one of the resistant strains. In vivo monooxygenase activity recorded in individual abdomens of flies showed that frequency distributions of ECOD activity in natural populations overlapped those of the resistant and laboratory strains, which were much narrower. Substantial and fast frequency changes (of the narrowness) that obtained in laboratory were related to either the time of rearing of 1 of the natural populations or selecting this population with an insecticide that has a toxicology correlated with both of the monooxygenase signs measured. Perspectives on using the CYP6A2 expression and ECOD activity for detecting a resistance mechanism by cytochrome P450 in field populations are discussed.

Animals↗
PubMed1994

Insecticide susceptibility in mosquitoes (Diptera: Culicidae) from French Polynesia.

Susceptibility to six organophosphate (OP), two pyrethroid (PY), and one carbamate (C) insecticides was investigated in Culex pipiens quinquefasciatus Say, Aedes aegypti (L.), and Aedes polynesiensis Marks larvae from the island of Tahiti. Cx. p. quinquefasciatus and Ae. aegypti were compared with susceptible reference strains treated simultaneously. A low, but significant, resistance to bromophos (4.6x), chlorpyrifos (5.7x), fenthion (2.4x), fenitrothion (5.0x), temephos (4.3x) and permethrin (2.1x) was found in Cx. p. quinquefasciatus, and to malathion (1.5x), temephos (2.3x), permethrin (1.8x) and propoxur (1.7x) in Ae. aegypti. Cx. p. quinquefasciatus was shown to possess over-produced esterases A2 and B2, which are known to be involved in resistance to OPs in other countries. Ae. polynesiensis was less resistant than the Ae. aegypti reference strain to all insecticides except temephos (1.8x) and permethrin (6.7x). To determine whether Ae. polynesiensis had developed resistance to these insecticides in Tahiti, a geographical survey covering 12 islands of the Society, Tuamotu, Tubuai, Marquesas, and Gambier archipelagoes was undertaken with three insecticides (temephos, deltamethrin, and permethrin). Two- to threefold variations in LC50S were observed among collections. Results are discussed in relationship to the level of insecticide exposure on the different islands.

Animals↗
PubMed1996

Yellowfever mosquito (Diptera:Culicidae) introduced into Landi Kotal, Pakistan, by tire importation.

Although the recent distribution of yellowfever mosquito, Aedes aegypti (L.), in Pakistan has been restricted to the port city of Karachi, adult and immature mosquitoes breeding in imported tires in warehouses at Landi Kotal (North-West Frontier Province) were identified as Ae. aegypti. The patterns of tire trade and the current disjunct distribution of Ae. aegypti indicated that the introduction into Landi Kotal may have been either from Karachi or India. Thermal fog application of pirimiphos-methyl and residual spray of malathion during 1993 reduced abundance in October-November. Living larvae or adults were not found during January 1994, apparently because of cold weather. However, Ae. aegypti reappeared during May-June 1994, most probably from eggs that overwintered. Population increased during late August when another round of spray using the same insecticides and fenthion as an additional larvicide again reduced abundance. Although this mosquito apparently has not spread into neighboring areas, its survival at Landi Kotal through all seasons despite control measures indicates its potential of becoming established in other areas of Pakistan.

Aedes↗