Metabolism of methiocarb and carbaryl by rat and human livers and plasma, and effect on their PXR, CAR and PPARα activities.

The oxidative, reductive, and hydrolytic metabolism of methiocarb and the hydrolytic metabolism of carbaryl by liver microsomes and plasma of rats or humans were examined. The effects of the metabolism of methiocarb and carbaryl on their nuclear receptor activities were also examined. When methiocarb was incubated with rat liver microsomes in the presence of NADPH, methiocarb sulfoxide, and a novel metabolite, methiocarb sulfone were detected. Methiocarb sulfoxide was oxidized to the sulfone by liver microsomes and reduced back to methiocarb by liver cytosol. Thus, the interconversion between methiocarb and the sulfoxide was found to be a new metabolic pathway for methiocarb by liver microsomes. The product of methiocarb hydrolysis, which is methylthio-3,5-xylenol (MX), was also oxidized to sulfoxide form by rat liver microsomes. The oxidations were catalyzed by human flavin-containing monooxygenase isoform (FMO1). CYP2C19, which is a human cytochrome P450 (CYP) isoform, catalyzed the sulfoxidations of methiocarb and MX, while CYP1A2 also exhibited oxidase activity toward MX. Methiocarb and carbaryl were not enzymatically hydrolyzed by the liver microsomes, but they were mainly hydrolyzed by plasma and albumin to MX and 1-naphthol, respectively. Both methiocarb and carbaryl exhibited PXR and PPARα agonistic activities; however, methiocarb sulfoxide and sulfone showed markedly reduced activities. In fact, when methiocarb was incubated with liver microsomes, the receptor activities were decreased. In contrast, MX and 1-naphthol showed nuclear receptor activities equivalent to those of their parent carbamates. Thus, the hydrolysis of methiocarb and carbaryl and the oxidation of methiocarb markedly modified their nuclear receptor activities.

Calix[4]arene triazole-linked pyrene: click synthesis, assembly on graphene oxide, and highly sensitive carbaryl sensing in serum.

A fluorescent calix[4]arene triazole-linked pyrene (CP) was carefully designed and synthesized via click chemistry. The modification of CP with graphene oxide (GO) by a simple non-covalent interaction strategy is presented. Further inspection by electrochemical impedance spectroscopy reveals that the CP-GO could exhibit a very high supramolecular recognition for carbaryl, in particular in serum samples with a nanomolar concentration detection. Additionally, it is easy to directly observe macroscopic recognition by the contact angle, and expand practical applications.

Use of in vitro data in developing a physiologically based pharmacokinetic model: Carbaryl as a case study.

In vitro-derived information has been increasingly used to support and improve human health risk assessment for exposure to chemicals. Physiologically based pharmacokinetic (PBPK) modeling is a key component in the movement toward in vitro-based risk assessment, providing a tool to integrate diverse experimental data and mechanistic information to relate in vitro effective concentrations to equivalent human exposures. One of the challenges, however, in the use of PBPK models for this purpose has been the need for extensive chemical-specific parameters. With the remarkable advances in in vitro methodologies in recent years, in vitro-derived parameters can now be easily incorporated into PBPK models. In this study we demonstrate an in vitro data based parameterization approach to develop a physiologically based pharmacokinetic and pharmacodynamic (PBPK/PD) model, using carbaryl as a case study. In vitro experiments were performed to provide the chemical-specific pharmacokinetic (PK) and pharmacodynamic (PD) parameters for carbaryl in the PBPK model for this compound. Metabolic clearance and cholinesterase (ChE) interaction parameters for carbaryl were measured in rat and human tissues. These in vitro PK and PD data were extrapolated to parameters in the whole body PBPK model using biologically appropriate scaling. The PBPK model was then used to predict the kinetics and ChE inhibition dynamics of carbaryl in vivo. This case study with carbaryl provides a reasonably successful example of utilizing the in vitro to in vivo extrapolation (IVIVE) approach for PBPK model development. This approach can be applied to other carbamates with an anticholinesterase mode of action as well as to environmental chemicals in general with further refinement of the current shortcomings in the approach. It will contribute to minimizing the need for in vivo human data for PBPK model parameterization and evaluation in human risk assessments.

Carbaryl and 1-naphthol tissue levels and related cholinesterase inhibition in male Brown Norway rats from preweaning to senescence.

Studies incorporating both toxicokinetic and dynamic factors provide insight into chemical sensitivity differences across the life span. Tissue (brain, plasma, liver) levels of the N-methyl carbamate carbaryl, and its metabolite 1-naphthol, were determined and related to brain and RBC cholinesterase (ChE) inhibition in the same animals. Dose-response (3, 7.5, 15, or 22.5 mg/kg, 40-45 min postdosing) and time course (3 or 15 mg/kg at 30, 60, 120, or 240 min postdosing) of acute effects of carbaryl (oral gavage) in preweanling (postnatal day [PND] 18) and adult male Brown Norway rats from adolescence to senescence (1, 4, 12, 24 mo) were compared. At all ages there were dose-related increases in carbaryl and 1-naphthol in the dose-response study, and the time-course study showed highest carbaryl levels at 30 min postdosing. There were, however, age-related differences in that the 1- and 4-mo rats showed the lowest levels of carbaryl and 1-naphthol, and PND18 and 24-mo rats had similar, higher levels. The fastest clearance (shortest half-lives) was observed in 1- and 4-mo rats. Carbaryl levels were generally higher than 1-naphthol in brain and plasma, but in liver, 1-naphthol levels were similar to or greater than carbaryl. Brain ChE inhibition closely tracked brain carbaryl concentrations regardless of the time after dosing, but there was more variability in the relationship between RBC ChE and plasma carbaryl levels. Within-subject analyses suggested somewhat more brain ChE inhibition at lower carbaryl levels only in the PND18 rats. These findings may reflect maturation followed by decline in kinetic factors over the life span.

Cholinesterase inhibition and depression of the photic after discharge of flash evoked potentials following acute or repeated exposures to a mixture of carbaryl and propoxur.

Previously, we reported that acute treatment with propoxur or carbaryl decreased the duration of the photic after discharge (PhAD) of flash evoked potentials (FEPs). In the current studies, we compared the effects of acute or repeated exposure to a mixture of carbaryl and propoxur (1:1.45 ratio; propoxur:carbaryl) on the duration of the PhAD and brain ChE activity in Long Evans rats. Animals were exposed (po) either to a single dose (0, 3, 10, 45 or 75 mg/kg), or 14 daily dosages (0, 3, 10, 30, 45 mg/kg), of the mixture. Acute and repeated treatment with 3mg/kg (or greater) of the mixture produced dose-related inhibition of brain ChE activity. Compared to controls, the PhAD duration decreased after acute administration of 75 mg/kg or repeated treatment with 30 mg/kg of the mixture. The linear relationship between the percent of control brain ChE activity and the PhAD duration was similar for both exposure paradigms. Dose-response models for the acute and repeated exposure data did not differ for brain ChE activity or the duration of the PhAD. Repeated treatment with the mixture resulted in slightly less (13-22%) erythrocyte ChE inhibition than acute exposure. Both acute and repeated treatment resulted in dose-additive results for the PhAD duration and less than dose-additive responses (6-16%) for brain ChE activity for the middle range of dosages. Acute treatment resulted in greater than dose-additive erythrocyte ChE inhibition (15-18%) at the highest dosages. In contrast, repeated treatment resulted in less than dose-additive erythrocyte ChE inhibition (16-22%) at the middle dosages. Brain and plasma levels of propoxur and carbaryl did not differ between the acute and repeated dosing paradigms. In summary, a physiological measure of central nervous system function and brain ChE activity had similar responses after acute or repeated treatment with the carbamate mixture, and brain ChE showed only small deviations from dose-additivity. Erythrocyte ChE activity had larger differences between the acute and repeated treatment paradigms, and showed slightly greater deviations from dose-additivity. Because these treatments utilized larger dosages than anticipated environmental exposures, concern for non-additive effects in humans is minimized. The small magnitude of the deviations from dose-additivity also suggest that in the absence of repeated exposure data, results from an acute study of readily reversible carbamate toxicity can be used to estimate the response to repeated daily exposures.

Relationship between brain and plasma carbaryl levels and cholinesterase inhibition.

Carbaryl is a N-methylcarbamate pesticide and, like others in this class, is a reversible inhibitor of cholinesterase (ChE) enzymes. Although studied for many years, there is a surprising lack of information relating tissue levels of carbaryl with ChE activity in the same animals. The present studies were undertaken to describe the dose-response relationship about 40 min (approximate time of maximal ChE inhibition) after oral treatment in adult, post-natal day (PND) 17, and PND11 rats. Additionally, the time-course of plasma ChE activity and carbaryl levels in adult rats was determined after a 30 mg/kg dosage of carbaryl. The time-course study found that carbaryl levels could be detected in plasma 1 h after dosing, but rapidly decreased below the level of quantitation by the 2 h time point. In the dose-response studies, treatment-related increases in plasma and brain carbaryl levels were observed 40 min after dosing. Plasma levels of carbaryl increased linearly, while brain levels appeared to asymptote after 75 mg/kg carbaryl. Plasma and brain levels of carbaryl appeared to be linearly related with a slope close to 1 after various dosages (range: 1-75 mg/kg) of carbaryl at the 40 min time point. Finally, the dose-related relationship between tissue levels of carbaryl and ChE activity was described using a first order exponential decay function with an asymptote. The parameters of this function did not appear to differ between adult, PND17, or PND11 rats. This indicates that age-related differences in brain ChE inhibition by carbaryl are unlikely to be the result of greater tissue levels of the pesticide in PND11 animals. These are the first studies to report the relationship between brain and plasma tissue levels of carbaryl and ChE activity on an individual animal basis. The results of these experiments will be useful to extend physiologically-based pharmacokinetic models for carbaryl and their application in risk assessment.

Metabolic profiles of serum from rats after subchronic exposure to chlorpyrifos and carbaryl.

Chlorpyrifos (CPF) and carbaryl (CAR) have been widely used in agricultural and domestic settings. Previous studies have demonstrated that CPF and CAR are generally neurotoxic to mammals, whereas the toxicities of these pesticides to other organs and their potential interactive effects remain unclear. The purpose of this study assessed the alterations of histopathology, biochemical parameters, and metabolic profiles of serum in rats following the treatment with CPF and CAR alone or in combination. No histopathological changes were observed in the liver and kidney tissues. Biochemical analysis of blood showed that alanine aminotransferase and total bilirubin in serum increased slightly in CPF-treated rats as compared to controls. Metabonomic analysis revealed alternations in a number of metabolites involving the metabolism of glucose, free fatty acids, and amino acids in liver mitochondria. The treatment of rats with CPF alone resulted in a decrease in lactate, low- and very low-density lipoprotein (LDL/VLDL), dimethylglycine (DMG), and aspartate. This was accompanied by an increase in isoleucine and leucine, 3-hydroxybutyrate (3-HB), N-acetylglycoprotein (NAC), acetone, succinate, glutamine, choline, creatine, glucose, and amino acids in a dose-dependent manner. Similarly, treatment with a high dose of CAR alone led to a decrease in DMG, aspartate, LDL/VLDL, and dimethylamine and an increase in taurine, glucose, and amino acids. The levels of lactate and LDL/VLDL decreased, while those of 3-HB, NAC, acetone, succinate, and glutamine elevated in the group of rats treated with a mixture of CPF and CAR as compared to the groups of CPF or CAR alone. Our results suggest that subchronic exposure to CPF and CAR alone, or in combination, could cause a disturbance in energy and fatty acid metabolism in the liver mitochondria of rats. Overall, we have shown that analysis of metabolic profiles can make exceptional contributions to the understanding of the individual or mutual effects following exposure to a low dose of pesticides.

Bayesian calibration of a physiologically based pharmacokinetic/pharmacodynamic model of carbaryl cholinesterase inhibition.

Carbaryl, an N-methyl carbamate (NMC), is a common insecticide that reversibly inhibits neuronal cholinesterase activity. The objective of this work was to use a hierarchical Bayesian approach to estimate the parameters in a physiologically based pharmacokinetic and pharmacodynamic (PBPK/PD) model from experimental measurements of carbaryl in rats. A PBPK/PD model was developed to describe the tissue dosimetry of carbaryl and its metabolites (1-naphthol and "other hydroxylated metabolites") and subsequently to predict the carbaryl-induced inhibition of cholinesterase activity, in particular in the brain and blood. In support of the model parameterization, kinetic tracer studies were undertaken to determine total radioactive tissue levels of carbaryl and metabolites in rats exposed by oral or intravenous routes at doses ranging from 0.8 to 9.2 mg/kg body weight. Inhibition of cholinesterase activity in blood and brain was also measured from the exposed rats. Markov Chain Monte Carlo (MCMC) calibration of the rat model parameters was implemented using prior information from literature for physiological parameter distributions together with kinetic and inhibition data on carbaryl. The posterior estimates of the parameters displayed at most a twofold deviation from the mean. Monte Carlo simulations of the PBPK/PD model with the posterior distribution estimates predicted a 95% credible interval of tissue doses for carbaryl and 1-naphthol within the range of observed data. Similar prediction results were achieved for cholinesterase inhibition by carbaryl. This initial model will be used to determine the experimental studies that may provide the highest added value for model refinement. The Bayesian PBPK/PD modeling approach developed here will serve as a prototype for developing mechanism-based risk models for the other NMCs.

Application of an in-line imprinted polymer column in a potentiometric flow-injection chemical sensor to the determination of the carbamate pesticide carbaryl in complex biological matrices.

A flow-injection biosensor-like system based on a nonenzymatic approach has been developed to determine the carbamate pesticide carbaryl in complex biological samples without lengthy and expensive extraction steps. Molecularly imprinted polymeric beads were used to immobilize carbaryl from biological samples. pH variation permitted the elution of carbaryl from the binding cavity to the flow cell. A pH electrode was used to detect changes in the charge of carbaryl in the sample solution resulting from the protonation and deprotonation of the molecule over different pH ranges. At pH 2.0, the secondary amine group is protonated, giving a (+1) charge to the carbaryl molecule. At pH 8.0, the ionized carbaryl loses a proton to become neutral, changing the local pH of the flow cell. The pH change at the flow cell generated by the deprotonation of carbaryl ion in alkaline medium was used to determine the carbaryl concentration. Parameters influencing the performance of the system were optimized for use in the detection procedure. The validated biosensor-like system had a carbaryl detection limit of 10.0 microg/mL and a response that was linear (r2>0.98) over the concentration range of 10.0-00 microg/mL.

Determination of carbofuran, carbaryl and their main metabolites in plasma samples of agricultural populations using gas chromatography-tandem mass spectrometry.

A gas chromatography-tandem mass spectrometric (GC-MS/MS) method has been developed for the determination of carbofuran (2,3-dihydro-2,2-dimethylbenzofuran-7-yl methylcarbamate), carbaryl (1-naphthyl-N-methylcarbamate) and their main metabolites in human blood plasma. Optimization of the isolation of the compounds from plasma matrix included the precipitation, denaturation and digestion of plasma proteins. Derivatization was achieved by the use of trifluoroacetic acid anhydride and was optimized for temperature, time and volume of derivatization agent. In the proposed method, a mild precipitation technique was applied using beta-mercaptoethanol and ascorbic acid in combination with solid-phase extraction technique using Oasis HLB (Hydrophobic Lipophilic Balance) cartridges for further clean up of samples. Carbamate linkage was not hydrolyzed to its phenol product, but both carbamate phenol and ketones were transformed into trifluoroacetyl derivatives in order to become volatile compounds and were determined using tandem mass spectrometry. The linearity of the method was shown for nine concentrations in the range of 0.50-250 ng mL(-1) in fortified plasma aliquots. Limits of detection (LODs) for all compounds ranged from 0.015-0.151 ng mL(-1). Inter-day and intra-day assays (RSD) for all compounds, at three concentration levels of 2.5, 25 and 100 ng mL(-1) (n=3) in fortified plasma samples were less than 18%. Accuracy (%E (r)) was calculated at three concentration levels, 8, 80 and 160 ng mL(-1) (n=3), and ranged from -12.0 to 15.0%. Matrix effect was evaluated so mean recoveries were calculated for all compounds and ranged from 81-107%. Specificity for the use of this method to biological monitoring studies was achieved including four main metabolites of CF, 1-naphthol and 2-naphthol from the naphthalene metabolism pathways, and both the parent compound of carbofuran and carbaryl. The proposed method was applied to plasma samples of pesticide users.

Pontine myelinolysis in a child with carbamate poisoning.

Carbamate and organophosphate pesticides are widely used all over the world. Poisoning with these substances may produce both immediate and delayed neurotoxic effects. We report the case of a 4-year-old boy who was admitted to the Pediatric Department of the Second University of Naples for evaluation of stupor, lethargy, severe hypotonia, generalized weakness of his arms and legs, ataxia, dysmetria, miosis, excessive salivation and tearing. The pesticide carbaryl (1-naphthyl-N-methylcarbamate) was identified in blood and urine samples. On admission, magnetic resonance imaging (MRI) was unremarkable; on day 11, MRI showed central pontine myelinolysis. The demyelination improved after 4 months and disappeared after 2 years. Various underlying and concomitant diseases have been described in children with central pontine myelinolysis but, to our knowledge, the finding of pontine myelinolysis after carbamate poisoning has not yet been described.

[Simultaneous determination of carbaryl and propanil in human serum and urine by on-line column-switching technique followed by automatic reversed-phase HPLC].

Carbaryl and propanil in human serum and urine were determined by automatic on-line column enrichment technique followed by reversed-phase HPLC with photometric detection. Human serum was filtered through a membrane filter (0.45 micron pore size) and an aliquot of 0.1 ml of the filtrate was diluted with water up to 1 ml. The solution of 0.8 ml was directly injected to automatic HPLC without any preparation. Urine was incubated with beta-glucuronidase/arylsulfate for 16 hours at 37 degrees C. The resultant solution was then filtered through a membrane filter and the filtrate was analyzed by the similar manner as serum. Carbaryl and propanil in the sample solution were concentrated on a pre-conditioned ODS mini-column. After washing the mini-column with 5% methanol, they were separated by an ODS analytical column (Cosmosil 5 C18-MS, 250 x 4.6 mm i.d.) with acetonitrile/water (30:70, v/v) eluent and detected with a UV detector. Carbaryl and propanil in serum were detected at 220 and 210 nm, respectively. On the other hand, in order to separate from blank peaks, carbaryl and propanil in urine were detected at 290 and 260 nm, respectively. The presented HPLC method requires neither manual procedure of solid-phase nor liquid-liquid extraction. Calibration curves for carbaryl and propanil were linear over the range of 5 ng/ml-2 micrograms/ml in both serum and urine. Real serum (ng/ml level) and urine (microgram/ml level) samples were analyzed by the presented HPLC method. Effect of seventeen pesticides on the determination of carbaryl and propanil were investigated. All pesticides did not interfere with the determination except for thiuram.

[Simultaneous determination of propanil, carbaryl and 3,4-dichloroaniline in human serum by HPLC with UV detector following solid phase extraction].

In case of poisoning by herbicide compounded with Propanil (DCPA) and Carbaryl (NAC), we attempted simultaneous solid-phase extractions of DCPA, NAC, and 3,4-dichloroaniline (DCA), a metabolite of DCPA, from the patient's serum, and quantitative analytical method using HPLC-UV detection. With this HPLC method, the quantitative detection limits in the serum are 0.005 microgram/ml for DCPA and DCA and 0.001 microgram/ml for NAC, and the UV spectra of all three compounds could easily be obtained using a diode-array detection limit of 0.05 microgram/ml. When the three compounds were added to serum at concentrations ranging from 0.1-10.0 micrograms/ml, the recovery rates were satisfactory at between 91.1% and 101.9%. On analysis of the serum of patient who had ingested Kusanon A Emulsion, the ingested substance apparently caused an increase in the DCA concentration, which led to the appearance of methemoglobinemia. The possibility that the DCA concentration might be used for prognostic purposes was suggested.

Dermal absorption and distribution of (14)C carbaryl in Wistar rats.

The level of (14)C carbaryl was determined in blood (leukocytes, erythrocytes, all blood cells, plasma) and organs (brain, heart, lungs, liver, spleen, skin at the site of exposure) of male Wistar rats after dermal administration. The application liquid was (14)C carbaryl solution in 96% ethyl alcohol. This preparation, possessing an activity of 670 kBq/ml, containing 1.67 mg of carbaryl, was applied to the skin of the tail according to Massmann's method in own modification. The amount of the preparation per 1 cm(2) of the tail skin was 0.19 mg of carbaryl (74.4 kBq). The tails of experimental rats were exposed to (14)C carbaryl by soaking for 4 h daily: once, twice or three times. Beta radiation from (14)C was measured in homogenized organs (brain, heart, lungs, liver, skin) and in blood by computer controlled Wallac scintillation counter Model 1409, using Multi Calc software. The dermal absorption of carbaryl at the site of exposure and in the surrounding area of about 2 cm was observed already during 4 hour exposure. Carbaryl reached plasma within 4 h of a single dermal exposure and penetrated into leukocytes, erythrocytes, heart, liver, lung, kidney and brain. The largest amount of (14)C carbaryl, about 2% of absorbed dose, was detected in liver

Reversed-phase high performance liquid chromatographic determination of carbaryl in postmortem specimens.

A reversed-phase high performance liquid chromatographic (HPLC) method has been developed for the analysis of carbaryl in postmortem samples. It involves the use of an aqueous acetonitrile mobile phase to give rapid, reproducible analyses and an external standard for quantitation. Two separate extraction procedures (Extrelut column and protein precipitation with acetonitrile) are described; each one is suited to a particular substrate and each one leads to an almost complete recovery of carbaryl from that substrate.

Influence of experimental hepatic impairment on the toxicokinetics and the anticholinesterase activity of carbaryl in the rat.

The blood kinetics of carbaryl were followed over 24 h after oral administration of 14C-carbaryl at 20 mg kg (0.17 mu Ci mg-1) in control animals and in animals with an altered liver function (70% hepatectomy or tranylcypromine treatment). The variations in the primary toxicity of carbaryl were assessed by measuring the inhibition of the plasma and erythrocyte cholinesterases and by evaluation of the lethal doses. The 14C radioactivity in the blood and, in parallel, cholinesterase inhibition were maintained at a higher level in animals with an altered hepatic function. A study of acute toxicity also showed a decrease of the LD50 (91 mg kg-1 with tranylcypromine, 342 mg kg-1 in the hepatectomized group) in the treated animals, with respect to the controls (585 mg kg-1). In all cases, tranylcypromine had a greater effect on blood kinetics, cholinesterase inhibition and LD50 than did 70% hepatectomy.

Percutaneous penetration of three insecticides in rats: a comparison of two methods for in vivo determination.

Percutaneous penetration of three insecticides was studied by two methods. The indirect (excretion analysis) and direct (skin patch removal) methods for determining penetration were compared in rats. Radiolabeled solutions of parathion, carbaryl, and DDT were applied to previously shaved rats at the rate of 4 micrograms/cm2. Recoveries of radioactivity in urine, feces, application site, and various tissues were measured at intervals over a 5-day period. Urinary excretion rates were corrected for incomplete excretion by intraperitoneal applications. In the 5 days following intraperitoneal administration, the urinary excretion of parathion and carbaryl was greater than 80% while less than 5% of DDT was excreted. A good correlation was found between the indirect and direct methods utilized to determine percutaneous absorption rates with the compounds tested at the later time intervals. All compounds showed more than 85% dermal penetration within 5 days. At the early time intervals (greater than 24 h), penetration by the direct method was significantly greater for parathion and carbaryl than by the indirect method.

Carbaryl tricompartmental toxicokinetics and anticholinesterase activity.

The blood kinetics of carbaryl and the inhibition of plasma acetylcholinesterases were followed for 24 h after administration of 20 mg/kg [14C]carbaryl (0.17 microCi/mg). The kinetics of the radioactivity attributed to unaltered carbaryl is bi-exponential whereas that of the total 14C activity is tri-exponential. The kinetics were treated with open 2- and 3-compartment models respectively. The exchange rate constants between the various compartments as well as the elimination constant (expressed in h-1) were found to be: k12 = 1.93, k21 = 1.18 and k10 = 2.46 (2-compartment model) and k'12 = 18.65, k'21 = 13.90, k'13 = 1.14 k'31 = 0.125 and k'10 = 0.672 (3-compartment model). The 3-compartment model demonstrates the persistence in the blood of 14C activity which correlated with plasma acetylcholinesterase inhibition.

High-performance liquid chromatographic determination of carbaryl and 1-naphthol in biological fluids.

A simple and sensitive method for the simultaneous analysis of carbaryl and 1-naphthol in whole blood by reversed-phase high-performance liquid chromatography and fluorescence detection is described. Spiked blood (heparinized) containing an internal standard was hemolyzed and extracted with ethyl acetate. After centrifugation the extractant was removed and taken to dryness. Reconstitution and subsequent high-performance liquid chromatography-fluorescence analysis yielded linear standard curves for carbaryl and 1-naphthol. Linear response vs. concentration profiles were obtained for carbaryl and 1-naphthol extracted from buffer solutions as well. A simple chemical hydrolysis study of carbaryl is included to illustrate the effectiveness of the extraction procedure and assay.