Determination of oxyfluorfen herbicide and oxyfluorfen amine residues in garbanzo beans by liquid chromatography

Oxyfluorfen and oxyfluorfen amine were determined by liquid chromatography (LC) with ultraviolet (UV) and photoconductivity detection (PCD). A simple extraction procedure acceptably recovered both analytes from garbanzo beans over a wide range of fortifications (0.05 to 20 ppm) (83 +/- 4 for oxyfluorfen; 85 +/- 4 for oxyfluorfen amine). Percent recoveries decreased slightly as the fortification level decreased. Both analytes could be determined simultaneously at a concentration greater than 0.2 ppm in garbanzo beans. Detection limits were 3 ng for oxyfluorfen and 100 ng for oxyfluorfen amine using LC/UV, and 12 ng for both oxyfluorfen and oxyfluorfen amine with LC/PCD. Different knitted reaction coils and photoreactors were evaluated. Photoproduct yields and identification were determined by ion chromatography. The LC/PCD method measures oxyfluorfen and oxyfluorfen amine separately and has a shorter analysis time, while the standard method using gas chromatography measures total residues and is more sensitive.     

Optimization of microwave-assisted extraction for the characterization of olive leaf phenolic compounds by using HPLC-ESI-TOF-MS/IT-MS(2)

In the present work, a simple and rapid method for the extraction of phenolic compounds from olive leaves, using microwave-assisted extraction (MAE) technique, has been developed. The experimental variables that affect the MAE process, such as the solvent type and composition, microwave temperature, and extraction time, were optimized using a univariate method. The obtained extracts were analyzed by using high-performance liquid chromatography (HPLC) coupled to electrospray time-of-flight mass spectrometry (ESI-TOF-MS) and electrospray ion trap tandem mass spectrometry (ESI-IT-MS(2)) to prove the MAE extraction efficiency. The optimal MAE conditions were methanol:water (80:20, v/v) as extracting solvent, at a temperature equal to 80 °C for 6 min. Under these conditions, several phenolic compounds could be characterized by HPLC-ESI-MS/MS(2). As compared to the conventional method, MAE can be used as an alternative extraction method for the characterization of phenolic compounds from olive leaves due to its efficiency and speed.     

Simultaneous determination of carbaryl, malathion, fenitrothion, and diazinon residues in sesame seeds (Sesamum indicum L.)

A method is described for the simultaneous determination of carbaryl (1-naphthyl methylcarbamate), malathion [diethyl (dimethoxythiophosphorylthio) succinate], fenitrothion (O,O-dimethyl O-4-nitro-m-tolyl phosphorothioate), and diazinon (O,O-diethyl O-2-isopropyl-6-methylpyrimidin-4-yl phosphorothioate) in sesame (Sesamum indicum L.) seeds. Sesame seeds were Soxhlet extracted with n-hexane, and the extract was subjected to a liquid-liquid partitioning and column cleanup to remove the oily coextractives prior to analysis by high performance liquid chromatography (HPLC). The mean percent recoveries (+/- standard deviations) from sesame seeds fortified with carbaryl (0.004 to 0.035 microgram/g), malathion (0.53 to 4.25 microgram/g), fenitrothion (0.22 to 1.78 microgram/g), and diazinon (0.54 to 4.35 microgram/g) were 83.3 +/- 5.7, 85.5 +/- 6.6, 85. 6 +/- 7.2, and 88.4 +/- 4.8, respectively. The method was used for the simultaneous analysis of carbaryl, malathion, fenitrothion, and diazinon residues in sesame seeds obtained from an Ethiopian field crop that had been treated with the pesticides during its growing period.     

Field-incurred fenitrothion residues in kakis: comparison of individual fruits, composite samples, and peeled and cooked fruits

Field trials have been carried out to determine the variability of residue levels of fenitrothion and its main metabolites fenitrothion-oxon and 3-methyl-4-nitrophenol in individual kaki fruits versus composite samples, in peel versus flesh, and in whole uncooked versus whole cooked fruits. Residue levels have been determined by gas chromatography with thermionic specific detection after extraction with ethyl acetate and without further cleanup. At harvest, residue levels of fenitrothion were below maximum residue levels (MRLs) and the two metabolites 3-methyl-4-nitrophenol and fenitrothion-oxon could be quantified with average amounts of 0.080 and 0.012 mg/kg, respectively. Levels of fenitrothion decreased 88% after peeling, whereas temperature did not result in a high variation. The ratios of the highest residue level in the individual fruits to the corresponding mean of residue levels in the composite samples for fenitrothion were <3. This value is lower than that recommended by the World Health Organization as default value for consumer risk assessment.     

Dissipation of pyrethroid residues in peppers,zucchinis, and green beans exposed to field treatments in greenhouses: evaluation by decline curves

Dissipation of seven pyrethroid insecticides under field conditions was evaluated on green beans, zucchinis, and peppers grown in experimental greenhouses (Almería, Spain). Pyrethroid residues were determined by high performance liquid chromatography using continuous on-line post-elution photoirradiation with fluorescence detection after dichloromethane extraction and cleanup on florisil phase cartridges. Mathematically defined decline curves were established by determining optimal relationships between pyrethroid residues and time. Different models were used to find these curves. The 1st-order model achieved the best adjustment to the experimental data in 42.9% of cases. The RF (root function) 1st-order model was the best in 33.3% of times. Each of the 1.5th- and 2nd-order models provided the best adjustment in a 9.5% of the cases. Finally, the RF 1.5th-order model was the most appropriate in only 4.8% of cases. Half-life times for these three vegetables were estimated from the optimal models. The preharvest intervals for the residues in these three vegetables was obtained, taking into account the maximum residue levels established by the existing legislation. They were all lower than the ones specified by the makers of commercial formulates, which ensures a safe enough consumption.     

Fate of mucilage cell wall polysaccharides during coffee fermentation.

Effects of a 20-h fermentation on cell wall polysaccharides from the mucilage of pulped coffee beans were examined and compared to those of unfermented beans, on alcohol insoluble residues (AIRs), their hot-water-soluble crude pectic substances (PECTs), and their hot-water-insoluble residues (RESs). Yields and compositions were very similar: AIRs, which consisted of approximately 30% highly methylated pectic substances, approximately 9% cellulose, and approximately 15% neutral noncellulosic polysaccharides, exhibited no apparent degradation. However, PECTs from fermented beans were shown to have undergone a slight reduction of their intrinsic viscosity and weight-average molecular weight by capillary viscosimetry and high-performance size-exclusion chromatography. After fermentation, hot-water-insoluble pectic substances of RES exhibited partial de-esterification. Removal of coffee bean mucilage by natural fermentation seems to result from a restricted pectolysis, the mechanism of which remains to be elucidated.     

Microwave-assisted extraction for the simultaneous determination of thiamethoxam, imidacloprid, and carbendazim residues in fresh and cooked vegetable samples

Microwave-assisted extraction (MAE) was carried out for the simultaneous determination of the insecticides thiamethoxam [(EZ)-3-(2-chloro-1,3-thiazol-5-ylmethyl)-5-methyl-1,3,5-oxadiazinan-4-ylidene(nitro)amine], imidacloprid [1-(6-chloro-3-pyridylmethyl)-N-nitroimidazolidin-2-ylideneamine], and the fungicide carbendazim (methyl benzimidazol-2-ylcarbamate) in vegetable samples. Five crop samples consisting of cabbage, tomatoes, chilies, potatoes, and peppers were fortified with the three pesticides and subjected to MAE followed by cleanup to remove coextractives prior to analysis by high-performance liquid chromatography. Using the selected microwave exposure time and power setting, the recoveries of the three pesticides from the fortified vegetable samples ranged from 68.1 to 106%. The corresponding recoveries for samples processed simultaneously but without microwave exposure ranged from 37.2 to 61.4%. The recoveries by MAE were comparable to those obtained by the conventional blender extraction technique. The precision of the MAE method was demonstrated by relative standard deviations of <7% for the three pesticides. The cooked cabbage and tomato samples showed no breakdown of the parent compounds, and the recoveries of three pesticides were comparable to those obtained with the uncooked samples.     

Analysis of trypsin inhibitors and lectins in white kidney beans (Phaseolus vulgaris, var. Processor) in a combined method.

Buffered saline extraction, affinity chromatography, and Folin-BSA protein assay were used consecutively to provide a combined method for analysis of trypsin inhibitors and lectins in white kidney beans (Phaseolus vulgaris, var. Processor). The method was tested by following the decrease of both antinutritional factors by germination of the beans for 7 days at 20 degrees C. Repeatability coefficients of variation were 2-7.4% for the trypsin inhibitors and 2.2-10% for the lectins. After 7 days of germination, trypsin inhibitors and lectins were reduced by 72 and 92%, respectively.     

Development of a monoclonal antibody-based cELISA for the analysis of sulfadimethoxine. 2. Evaluation of rapid extraction methods and implications for the analysis of incurred residues in chicken liver tissue

Several rapid extraction methods were evaluated for use with a monoclonal antibody-based competitive inhibition ELISA (cELISA) to detect sulfadimethoxine (SDM) in chicken liver tissue. These methods included extraction of the samples with (1) aqueous buffer with or without ultrafiltration, (2) acetonitrile/water, (3) methanol/water, or (4) acetone. The organic extraction methods were evaluated with or without solvent evaporation prior to dilution into assay buffer for the cELISA. The aqueous-based extraction methods were compatible with the cELISA. However, of the organic extraction methods, only the acetone liver extract with solvent evaporation prior to analysis was compatible with the cELISA. The cELISA method coupled to aqueous- or acetone-based sample extraction as well as an HPLC method was evaluated for the analysis of chicken liver tissues fortified with SDM at levels from 0.2 to 0.025 ppm. Mean SDM recoveries for the HPLC method and for the cELISA method using samples prepared by aqueous extraction, aqueous extraction and ultrafiltration, or acetone extraction, evaporation, and reconstitution were 68.9, 95.7, 60.1, and 52.5%, respectively. For the analysis of samples obtained from an SDM incurred residue study, HPLC and cELISA analysis of the same organic extract gave results that were highly correlated (R(2) = 0.976; p < 0.0001). However, results obtained from the analysis of aqueous extracts by cELISA did not correlate well with those obtained by HPLC (R(2) = 0.61, p > 0. 0006). This was attributed to the coextraction of cross-reactive SDM-related residues that were not quantified by the HPLC method. The presence of these residues should be considered during data interpretation when ELISA methods coupled with rapid aqueous extraction of samples are used in SDM residue monitoring programs.     

HPLC method for the quantification of procyanidins in cocoa and chocolate samples and correlation to total antioxidant capacity.

Monomeric and oligomeric procyanidins present in cocoa liquors and chocolates were separated and quantified in four different laboratories using a normal-phase high-performance liquid chromatography (HPLC) method with fluorescence detection. Procyanidin standards through decamers were obtained by extraction from cocoa beans, enrichment by Sephadex LH-20 gel permeation chromatography, and final purification by preparative normal-phase HPLC. The purity of each oligomeric fraction was assessed using HPLC coupled to mass spectrometry. A composite standard was then prepared, and calibration curves were generated for each oligomeric class using a quadratic fit of area sum versus concentration. Results obtained by each of the laboratories were in close agreement, which suggests this method is reliable and reproducible for quantification of procyanidins. Furthermore, the procyanidin content of the samples was correlated to the antioxidant capacity measured using the ORAC assay as an indicator for potential biological activity.     

Simplified Dilute Acetic Acid Based Extraction Procedure for Fractionation and Analysis of Wheat Flour Protein by Size Exclusion HPLC and Flow Field‐Flow Fractionation

A simple, highly efficient and reproducible two‐step extraction procedure using dilute acetic acid without (AN) and then with sonication (AS) has been developed for the fractionation of wheat flour protein. Approximately 97% of total protein was extracted from a Canadian hard red spring wheat flour; an additional 1.2% protein could be recovered by further extraction with 1% DDT and 50% 1‐propanol (AR). Size‐exclusion HPLC (SE‐HPLC) and flow field‐flow fractionation (flow FFF) showed that the AN extract, which accounted for most of the total extractable protein (AN + AS + AR), consisted primarily of monomeric protein. The AS extract was composed primarily of polymeric proteins. Flow FFF showed that AN polymeric protein, including that eluting at the SE‐HPLC void volume, showed smaller Stokes diameters than AS polymeric protein. Flow FFF profiles of AS SE‐HPLC subfractions showed that the void volume subfraction contained monomeric and small polymeric protein in addition to large polymeric protein, indicating formation of larger complexes through interaction between some or all of the components. AN and AS extracts, as well as SE‐HPLC and flow FFF fractions thereof, showed a fairly wide range of values among 12 Canadian hard red and white spring wheat cultivars. The proportion of total protein in the AS extract and in the larger sized polymeric protein fractions from SE‐HPLC and flow FFF were highly positively correlated to farinograph mixing time.     

Preparation of ferulic acid from agricultural wastes: its improved extraction and purification

Ferulic acid (FA) is a phenolic antioxidant present in plants, which is widely used in the food and cosmetic industry. In the present study, various agricultural wastes such as maize bran, rice bran, wheat bran, wheat straw, sugar cane baggasse, pineapple peels, orange peels, and pomegranate peels were screened for the presence of esterified FA (EFA). Among the sources screened, maize bran was found to contain the highest amount of EFA. Pineapple peels, orange peels, and pomegranate peels were also found to contain traces of EFA. Alkaline extraction of EFA from maize bran was carried out using 2 M NaOH. Response surface methodology (RSM) was used for optimization of EFA extraction, which resulted in a 1.3-fold increase as compared to the unoptimized conventional extraction technique. FA was analyzed by means of high-performance liquid chromatography (HPLC). Purification was carried out by adsorption chromatography using Amberlite XAD-16 followed by preparative high-performance thin-layer chromatography (HPTLC). The recovery of Amberlite XAD-16 purified FA was up to 57.97% with HPLC purity 50.89%. The fold purity achieved was 1.35. After preparative HPTLC, the maximum HPLC purity obtained was 95.35% along with an increase in fold purity up to 2.53.     

Determination of triazine and chloroacetanilide herbicides in soils by microwave-assisted extraction (MAE) coupled to gas chromatographic analysis with either GC-NPD or GC-MS

A simple and rapid method based on microwave-assisted extraction (MAE) coupled to gas chromatographic analysis was developed for the analysis of triazine (atrazine, cyanazine, metribuzine, simazine and deethylatrazine, and deisopropylatrazine) and chloroacetanilide (acetochlor, alachlor, and metolachlor) herbicide residues in soils. Soil samples are processed by MAE for 5 min at 80 degrees C in the presence of acetonitrile (20 mL/sample). Mean recovery values of most solutes are >80% in the 10 to 500 microg/kg fortification range with respective RSDs (relative standard deviations) < 20%. The limits of quantification (LOQ) and limits of detection (LOD) are 10 and 1 to 5 microg/kg, respectively. The method was validated with two types of soils containing 1.5 and 3.0% organic matter content, respectively; no statistically significant differences were found between solute recovery values from the two types of soils. The solute mean recovery values from freshly spiked (24 h aging) and spiked samples stored refrigerated for one week before processed were also not statistically different. Residue levels determined in field weathered soils were higher when soils were processed by MAE than with a comparison method based on flask-shaking of soil suspensions overnight. Extracts were analyzed by a gas chromatographic system equipped either with a thermionic (GC-NPD) or a mass spectrometric detector (GC-MS).     

Identification of ethyl 2-sulfanylacetate as an important off-odor compound in white wines

A number of Sauvignon blanc wines made from hard pressed juices in an inert atmosphere (nitrogen) or in contact with oxygen were identified as having heavy off-flavors to varying degrees. Samples were extracted and subjected to time-based HPLC fractionation. The fractions were assessed by a sensory panel and those with unpleasant, irritating, off-odors were re-extracted. The extracts evaluated by gas chromatography coupled with olfactometry revealed a number of odoriferous zones, including one with an off-odor similar to the one perceived in two HPLC fractions. The odor was less intense in fractions previously supplemented with copper sulfate, suggesting that the compound(s) responsible were possibly thiol-related. A selective thiols extraction protocol and the analysis of the extract by gas chromatography coupled with mass spectrometry identified a new potent thiol in these wines. The compound responsible for the odoriferous zone, ethyl 2-sulfanylacetate (1), had an odor reminiscent of baked beans and Fritillaria meleagris bulbs. Its perception threshold was determined and sensory studies using graduated supplementation in dry white wines demonstrated its contribution to the off-odor observed in dry white wines.     

Effects of Temperature,Sonication Time,and Power Settings on Size Distribution and Extractability of Total Wheat Flour Proteins as Determined by Size‐Exclusion High‐Performance Liquid Chromatography

The size‐exclusion (SE) HPLC profile of total protein extract obtained by sonication of flour samples at ambient temperature showed marked instability on reinjection. Instability was related to the presence of flour proteases that were inactivated by thermal treatment of flour samples at 60°C. Extraction of flour protein by sonication was a function of ultrasonic energy (sonication time × power product) delivered to flour sample. As protein solubility increased, the proportions of the earliest eluted SE‐HPLC fractions (F1 and F2) increased. Oversonication of proteins evidenced by a decrease in F1 amount at the benefit of F2 occurred below the sonication energy level needed for total protein extraction. Ultrasonic energy level was adjusted to allow total protein extraction while limiting oversonication. The sonication procedure was applied on 27 flour samples of contrasting dough strength to extract total proteins. Absolute amount of protein extractable by sonication, determined from SE‐HPLC area, was strongly correlated with flour protein content. Very significant and equivalent relationships were found between alveographic W index and absolute amount of either unextractable protein extract or F1 of SE‐HPLC profile from total protein extract.     

Determination of organochlorine and organophosphorus pesticide residues in eggs using a solid phase extraction cleanup

A multiresidue solid phase extraction (SPE) method for the isolation and subsequent gas chromatographic determination of nonpolar organochlorine and polar organophosphorus pesticide residues in eggs is described. The method uses an acetonitrile extraction followed by an SPE cleanup using graphitized carbon black and aminopropyl SPE columns. Organophosphorus pesticides are determined by gas chromatography with flame photometric detection. After further cleanup of the extract using Florisil SPE columns, organochlorine pesticides are determined by gas chromatography with electron capture detection. Studies were performed using eggs containing both fortified and incurred pesticide residues. The average recoveries were 86-108% for 8 fortified organochlorine pesticide residues and 61-149% for 28 fortified organophosphorus pesticide residues.     

Determination of niclosamide residues in rainbow trout (Oncorhynchus mykiss) and channel catfish (Ictalurus punctatus) fillet tissue by high-performance liquid chromatography

Bayluscide [the ethanolamine salt of niclosamide (NIC)] is a registered piscicide used in combination with 3-(trifluoromethyl)-4-nitrophenol (TFM) to control sea lamprey populations in streams tributary to the Great Lakes. A high-performance liquid chromatography (HPLC) method was developed for the determination of NIC residues in muscle fillet tissues of fish exposed to NIC and TFM during sea lamprey control treatments. NIC was extracted from fortified channel catfish and rainbow trout fillet tissue with a series of acetone extractions and cleaned up on C(18) solid-phase extraction cartridges. NIC concentrations were determined by HPLC with detection at 360 and 335 nm for rainbow trout and catfish, respectively. Recovery of NIC from rainbow trout (n = 7) fortified at 0.04 microg/g was 77 +/- 6.5% and from channel catfish (n = 7) fortified at 0.02 microg/g was 113 +/- 11%. NIC detection limit was 0.0107 microg/g for rainbow trout and 0.0063 microg/g for catfish. Percent recovery of incurred radioactive residues by this method from catfish exposed to [(14)C]NIC was 89.3 +/- 4.1%. Percent recoveries of NIC from fortified storage stability tissue samples for rainbow trout (n = 3) analyzed at 5 and 7.5 month periods were 78 +/- 5.1 and 68 +/- 2.4%, respectively. Percent recoveries of NIC from fortified storage stability tissue samples for channel catfish (n = 3) analyzed at 5 and 7.5 month periods were 88 +/- 13 and 76 +/- 21%, respectively.     

An LC/MS/MS method for improved quantitation of the bound residues in the tissues of animals orally dosed with [(14)C]Benomyl

Livers of goats orally dosed with [phenyl(U)-(14)C]benomyl contained radioactive residues which were not extractable using conventional, solvent-based extraction methods. We report a new residue method capable of enhanced extraction of benomyl-derived residues with selective and sensitive quantitation capability for methyl 4-hydroxybenzimidazol-2-ylcarbamate (4-HBC), methyl 5-hydroxybenzimidazol-2-ylcarbamate (5-HBC), and methyl benzimidazol-2-ylcarbamate (MBC). This method involves rigorous Raney-nickel reduction of hypothesized thioether bonds between benomyl residues and polar cellular components. Following acidic dehydration (desulfurization), the polar benomyl-derived residues are extracted into ethyl acetate and analyzed by LC/MS/MS. We have shown this method to be superior to alternative extraction approaches. When applied to goat liver tissue containing [phenyl(U)-(14)C]benomyl-bound residues, the extraction efficiency of total radioactive residues was approximately 30%, and the major benomyl-derived residue was 5-HBC (91-95% of extractable residue) with minor levels of carbendazim (MBC) (5-9%). HPLC/LSC data were consistent with the LC/MS/MS data. The overall method satisfies U.S. regulatory requirements in extraction efficiency, selectivity in detection, and limits of quantitation for benomyl-bound residues.     

Rapid and sensitive determination of 4-nitrophenol, 3-methyl-4-nitrophenol, 4,6-dinitro-o-cresol, parathion-methyl, fenitrothion, and parathion-ethyl by liquid chromatography with electrochemical detection

Liquid chromatography with electrochemical detection has been used to determine various nitropesticides, DNOC, fenitrothion, and parathion (methyl and ethyl), and some of their main metabolites, 4-nitrophenol for parathion (methyl and ethyl) and 3-methyl-4-nitrophenol for fenitrothion, by using indirect detection. Analysis of them in river water samples has been performed without a preconcentration step. The recovery efficiencies of the tested compounds yielded values between 96 and 112% at the fortification level of 0.5 ppb in a river water sample, and their relative standard deviations were between 1 and 15%. The detection limits of these compounds ranged between 0.05 and 0.14 ppb.     

Determination of the 2H/1H and 15N/14N ratios of Alkylpyrazines from coffee beans (Coffea arabica L. and Coffea canephoravar. robusta) by isotope ratio mass spectrometry

The delta15N(AIR) and delta2H(VSMOW) data for several alkylpyrazines formed during the roasting process of coffee are reported. Samples of commercially available roasted (n = 9) as well as self-roasted (n = 8) coffee beans (Coffea arabica L. and Coffea canephora var. robusta) of different origins were investigated. By use of extracts prepared by simultaneous distillation extraction (SDE) and subsequently fractionated by liquid chromatography on silica gel, on-line capillary gas chromatography-isotope ratio mass spectrometry was employed in the combustion (C) and pyrolysis (P) modes (HRGC-C/P-IRMS) to determine the delta15N(AIR) and delta2H(VSMOW) values, respectively. In addition to the constituents of coffee beans, data for commercial synthetic alkylpyrazines and substances declared to be "natural" were determined. The delta15N(AIR) data for coffee alkylpyrazines under study-2-ethyl-5-methylpyrazine (1) and 2-ethyl-6-methylpyrazine (2) (measured as sum 1/2), 2-ethyl-3-methylpyrazine (3), 2-methylpyrazine (4), 2,5-dimethylpyrazine (5) and 2,6-dimethylpyrazine (6) (measured as sum 5/6), and 2,3-dimethylpyrazine (7), as well as 2,3,5-trimethylpyrazine (8)-varied in the range from +8.3 to -10.2 per thousand, thus revealing their biogeneration from amino acids (delta15N(AIR) ranging from +8 per thousand to -10 per thousand). The delta2H(VSMOW) values were determined in the range from -5 per thousand to -127 per thousand. Owing to the analytical differentiation observed between coffee alkylpyrazines and synthetic/"natural" samples of 3, 4, and 7, authenticity assessment of coffee-flavored products seems to be promising, provided that extended data will be available in the future. In the literature, there were no IRMS data available for the alkylpyrazines (1-8) under study.