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This study employed direct UV-ABC photolysis and the UV-ABC/H 2 O 2 process to investigate the degradation of tolfenamic acid (TA), a common anti-inflammatory drug used in both human and veterinary medicine. A 2 3 factorial design with added center point was used to evaluate the effect of three independent variables-namely, H 2 O 2 concentration ([H 2 O 2 ]), TA concentration ([TA]), and experiment time (time)-on TA degradation and H 2 O 2 photolysis during UV-ABC/H 2 O 2 treatment using a high-pressure mercury vapor lamp (photon flux of 2.6307 × 10 4 J s -1 ) as the UV irradiation source. The responses yielded similar values, revealing a linear behavior, with correlation coefficients R = 0.9968 and R adj = 0.9921 for TA degradation and R = 0.9828 and R adj = 0.9570 for H 2 O 2 photolysis. The most efficient combination of variables was [H 2 O 2 ] = 255 mg L -1 and [TA] = 25 mg L -1 , resulting in 100% TA degradation and 98.87% H 2 O 2 photolysis by 90 min of treatment. Additionally, the second-order kinetic constant of the reaction between TA and HO ● was determined using a competitive kinetic model, employing 2,4-dichlorophenoxyacetic acid (2,4D) as the reference compound. The kinetic constant was 1.9 × 10 10 M -1 s -1 in alkaline medium. TA degradation by direct photolysis generated quinone imines as by-products, responsible for the formation of a dark red "internal filter" that increased the value of acute toxicity to Artemia salina. The UV-ABC/H 2 O 2 process did not promote formation of quinone imines by 90 min of treatment and therefore did not increase acute toxicity values. Several by-products generated during TA degradation were identified and possible degradation pathways for the UV-ABC and UV-ABC/H 2 O 2 processes were proposed., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The crystal structure of fenamic acid-acridine complex is determined by X-ray diffraction. The strong OHN hydrogen bond linking the complex components and other interactions responsible for packing of the molecules into a crystal are investigated within the Quantum Theory of Atom in Molecule theory. The crystal structure is compared with the structure optimized at B3LYP/6-311++G** level and with the theoretical structures optimized under systematically changed pressure. Analysis of the lattice constants, hydrogen bond lengths, and angles of the inter- and intramolecular hydrogen bond under compression is performed. The structural transformation observed at 5 GPa is connected with a change in the intermolecular OHN hydrogen bond. The proton shifts to acceptor and a new interaction in the crystal appears., (Copyright © 2016 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.)

To study the dissipation rates and final residual levels of chlorantraniliprole and thiamethoxam in maize straw, maize, and soil, two independent field trials were conducted during the 2014 cropping season in Beijing and Anhui Provinces of China. A 40% wettable powder (20% chlorantraniliprole + 20% thiamethoxam) was sprayed onto maize straw and soil at an application rate of 118 g of active ingredient per hectare (g a.i.ha(-1)). The residual concentrations were determined by ultra-high-performance liquid chromatography-tandem mass spectrometry. The chlorantraniliprole half-lives in maize straw and soil were 9.0-10.8 and 9.5-21.7 days, respectively. The thiamethoxam half-lives in maize straw and soil were 8.4-9.8 and 4.3-11.7 days, respectively. The final residues of chlorantraniliprole and thiamethoxam in maize straw, maize, and soil were measured after the pesticides had been sprayed two and three times with an interval of 7 days using 1 and 1.5 times the recommended rate (72 g a.i. ha(-1) and 108 g a.i. ha(-1), respectively). Representative maize straw, maize, and soil samples were collected after the last treatment at pre-harvest intervals of 7, 14, and 28 days. The chlorantraniliprole residue was below 0.01 mg kg(-1) in maize, between 0.01 and 0.31 mg kg(-1) in maize straw, and between 0.03 and 1.91 mg kg(-1) in soil. The thiamethoxam residue concentrations in maize, maize straw, and soil were <0.01, <0.01, and 0.01-0.03 mg kg(-1), respectively. The final pesticide residues on maize were lower than the maximum residue limit (MRL) of 0.02 mg kg(-1) after a 14-day pre-harvest interval. Therefore, a dosage of 72 g a.i. ha(-1) was recommended, as it can be considered safe to human beings and animals.

While much is known about the benefits of oat fiber, the relevance of phenolics from oat based food products remains unclear. To gain insight into the relevance of phenolics from consumer oat products, the content of phenolics and avenanthramides (AVEs) was followed in 10 oat cultivars as well as milled oat ingredients and ready to eat (RTE) consumer products (puffed cereals and snack bars). Free phenolic content ranged from 17.2 to 228.1 μg g(-1) DW with AVEs accounting for 57.3-90.6% of the total free phenolics. Bound phenolic content ranged from 141.4 to 680.9 μg g(-1) DW with ferulic acid accounting for 62-94% of the bound phenolics. Select oat groats were ground to flour and prepared as wet cooked porridges (∼20% oat flour in boiling water) or introduced as RTE products into a three stage in vitro digestion to determine phenolic bioaccessibility. The relative bioaccessibility for wet cooked porridges ranged from 0.3-2.6% and from 2.9-28.8% for individual phenolic acids and AVEs, respectively. Puffed oat cereal had significantly higher bioaccessibility compared to matching wet-cook porridge made from the same oat flour (e.g. 83.8% versus 19.1% for AVE A; p < 0.05). Intestinal uptake of bioaccessible oat phenolics from digesta was confirmed by experiments with Caco-2 human intestinal cells. The overall intestinal uptake of oat phenolics was low, ranging between 0.16% and 2.71% across digesta from all oat products. In conclusion, these experiments demonstrate that phenolics naturally present in whole grain oats are recovered well through traditional grain processing, milling and food processing. Furthermore, processing of oat cultivars into a RTE cereal may have a positive impact on the digestive release and bioaccessibility of oat phenolics in the upper GI tract.

Transcriptional co-activator with PDZ-binding motif (TAZ) plays versatile roles in cell proliferation and differentiation. It is phosphorylated by large tumor suppressor kinases, the core kinases of the tumor-suppressive Hippo pathway. Phosphorylation induces the cytoplasmic accumulation of TAZ and its degradation. In human cancers, the deregulation of the Hippo pathway and gene amplification enhance TAZ activity. TAZ interacts with TEA domain family members (TEAD), and upregulates genes implicated in epithelial-mesenchymal transition. It also confers stemness to cancer cells. Thus, TAZ activation provides cancer cells with malignant properties and worsens the clinical prognosis. Therefore, TAZ attracts attention as a therapeutic target in cancer therapy. We applied 18 606 small chemical compounds to human osteosarcoma U2OS cells expressing GFP-fused TAZ (GFP-TAZ), monitored the subcellular localization of GFP-TAZ, and selected 33 compounds that shifted GFP-TAZ to the cytoplasm. Unexpectedly, only a limited number of compounds suppressed TAZ-mediated enhancement of TEAD-responsive reporter activity. Moreover, the compounds that weakened TEAD reporter activity did not necessarily decrease the unphosphorylated TAZ. In this study, we focused on three compounds that decreased both TEAD reporter activity and unphosphorylated TAZ, and treated several human cancer cells with these compounds. One compound did not show a remarkable effect, whereas the other two compounds compromised the cell viability in certain cancer cells. In conclusion, the GFP-TAZ-based assay can be used as the first screening for compounds that inhibit TAZ and show anticancer properties. To develop anticancer drugs, we need additional assays to select the compounds., (© 2016 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.)

Chlorantraniliprole 0.4 % GR has been in use for managing early shoot borer and top borer of sugarcane. Persistence and dissipation kinetics of granular formulation of chlorantraniliprole were studied in the soil of tropical sugarcane ecosystem by employing simple and sensitive analytical method. Limit of quantification of the method was 0.01 mg/kg and the recovery of chlorantraniliprole was in the range of 92.3-99.7 % with RSD of 1.14-3.0 %. The initial deposit of chlorantraniliprole in the soil was 0.513 and 1.031 mg/kg for the recommended (75 g a.i./ha) and double the recommended (150 g a.i./ha) doses, respectively. The residues were quantified up to 30 days after treatment irrespective of the doses applied. Half-life (t 1/2) was 6.60 and 6.73 days, respectively, for recommended and double the recommended doses of chlorantraniliprole.

Polyethylene terephthalate (PET) bottles are widely used as packaging material for natural mineral water. However, trace levels of acetaldehyde can migrate into natural mineral water during the shelf life and might influence the taste of the PET bottled water. 2-Aminobenzamide is widely used during PET bottle production as a scavenging agent for acetaldehyde. The aim of this study was the determination of the migration kinetics of 2-aminobenzamide into natural mineral water as well as into 20% ethanol. From the migration kinetics, the diffusion coefficients of 2-aminobenzamide in PET at 23 and 40°C were determined to be 4.2 × 10(-)(16) and 4.2 × 10(-)(15) cm(2) s(-1), respectively. The diffusion coefficient for 20% ethanol at 40°C was determined to be 7.7 × 10(-)(15) cm(2) s(-1), which indicates that 20% ethanol is causing swelling of the PET polymer. From a comparison of migration values between 23 and 40°C, acceleration factors of 9.7 when using water as contact medium and 18.1 for 20% ethanol as simulant can be derived for definition of appropriate accelerated test conditions at 40°C. The European Union regulatory acceleration test based on 80 kJ mol(-1) as conservative activation energy overestimates the experimentally determined acceleration rates by a factor of 1.6 and 3.1, respectively.

Dissipation and decontamination of chlorantraniliprole (Coragen 18.5 SC) in brinjal and okra fruits were studied following field application at single and double doses of 30 and 60 g ai ha(-1), and the residues of the insecticide was estimated using LC-MS/MS. Initial residues of chlorantraniliprole at single and double doses on the fruits of brinjal were 0.72 and 1.48 mg kg(-1), while on okra fruits, the residues were 0.48 and 0.91 mg kg(-1), respectively. The residues reached below detectable level of 0.01 mg kg(-1) on the 10th day. Half-life of chlorantraniliprole at 30 and 60 g ai ha(-1) on brinjal was 1.58 and 1.80 days with the calculated waiting period of 0.69 and 2.38 days, whereas on okra, the values were 1.60 and 1.70 and 0 and 1.20 days, respectively. The extent of removal of chlorantraniliprole using simple decontaminating techniques at 2 h and 3 days after spraying was 40.99-91.37% and 29.85-89.12%, respectively, from brinjal fruits and 47.78-86.10% and 41.77-86.48%, respectively, from okra fruits.

Two accurate, selective and precise chromatographic methods, namely thin-layer chromatography (TLC)-densitometric method and reversed phase high-performance liquid chromatography (RP-HPLC) method, were developed and validated for the simultaneous determination of tolfenamic acid (TOL) and its two major impurities, 2-chlorobenzoic acid (CBA) and 3-chloro-2-methylaniline (CMA), which are also reported to be its related substances. The developed TLC-densitometric method depended on separation and quantitation of the studied drugs on silica gel 60F254 TLC plates. Hexane:chloroform:acetone:acetic acid (75:25:20:0.1, v/v/v/v) was used as a developing system and the separated bands were UV-scanned at 240 nm. Linear relationships were obtained in the range of 10-100 µg band(-1) for the drug and in the range of 0.1-1 µg band(-1) for the studied impurities. The developed RP-HPLC depended on chromatographic separation of the studied drugs on a C18 column using 0.05 M KH2PO4 buffer (pH 3):acetonitrile (45:55, v/v) as a mobile phase delivered at constant flow rate of 1 mL min(-1) with UV detection at 230 nm. Calibration curves for TOL and the two impurities were constructed over the concentration ranges of 10-100 µg mL(-1) for TOL and 0.01-0.1 µg mL(-1) for both CBA and CMA. Factors affecting the developed methods have been studied and optimized. Further, methods validation has been carried out according to International Conference on Harmonization guidelines. The proposed methods were successfully applied for determination of the studied drug in its bulk powder and in pharmaceutical formulation. The methods showed no significant difference when compared with the reported RP-HPLC one. The developed methods have advantages of being more sensitive and specific than the published methods., (© The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Thiamethoxam and chlorantraniliprole insecticides have been important tools for controlling pests in rice. However, food safety issues related to pesticide residues are important to consider with a food crop such as rice. Therefore, the objective of this study was to analyze thiamethoxam and chlorantraniliprole residues in rice hull, bran, and polished rice grains. The study was conducted during the 2012 cropping season at the Texas A&M Agrilife Research, David R. Wintermann Rice Research Station, near Eagle Lake, TX, USA. Rice was planted on May 5, 2012, using the cultivar 'Presidio'. Pesticide applications were performed at 5, 15, 25, and 35 days after flowering (DAF) using 1 and 2 times the recommended rate of 30 g active ingredient (ai) ha(-1) for thiamethoxam and 30 g ai ha(-1) for chlorantraniliprole. Sequentially, two treatments using the insecticides at recommended rate were applied at 5 and 25 DAF and at 5, 25, and 35 DAF. Insecticide residues were analyzed in different sample fractions: rice hull, bran, and polished rice grains. The samples were subjected to extraction using an accelerated solvent extraction (ASE) technique. Sample aliquots were analyzed using ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), with a limit of quantification (LOQ) of 5 × 10(-5) mg kg(-1). Residues of thiamethoxam and chlorantraniliprole were detected in rice hull, bran, and polished rice grains, and the quantified values were greater in hull and in rice bran.

The aim of this study was to identify the chemical structure of the photodegradation products of furosemide in a water-acetonitrile mixture (1 : 1). Furosemide solution was irradiated with a D65 fluorescent lamp and the products were isolated by preparative HPLC. The fractions were evaporated to dryness in vacuo. The purity of the photodegradation products was measured by HPLC. The purity of products 1, 3, and 4 was greater than 90%, whereas that of product 2 was 13%, therefore, photodegradation product 2 was unstable. We identified photodegradation products 1 and 3 as 4-chloro-5-sulfamoylanthranilic acid and 4-hydroxy-N-furfuryl-5-sulfamoylanthranilic acid, respectively, by LC/MS and NMR. Additionally, we assumed that photodegradation product 4 was methyl 2-((furan-2-ylmethyl)amino)-4-hydroxy-3-(methyleneamino)-5-sulfamoylbenzoate by LC/MS and NMR. This showed that furosemide underwent hydrolysis and substitution, and reacted with the acetonitrile under the light of a D65 fluorescent lamp. We were furthermore able to determine the elution times of the photodegradation products of furosemide by applying the Japanese Pharmacopoeia chromatographic method for related substances to the isolated products.

Photodegradation of [(14)C]-chlorantraniliprole (CLAP) and [(14)C]-cyantraniliprole (CNAP) was investigated in sterile buffer solutions, in natural water, and on soil surfaces. Both compounds displayed rapid degradation in aqueous buffers when exposed to light at concentrations which could result from direct overspray to a shallow water body. While the main products observed had analogous structures, a substantial difference was noted in the rate of degradation of the two compounds despite minimal differences in their structures. Transformations observed were primarily intramolecular rearrangements and degradations resulting from addition of hydroxyl radicals leading to molecular cleavage. Some of the degradation products were transient, and several degradates had isomeric molecular compositions. The sequence of transformations was established definitively with the help of kinetics modeling. Utility of kinetics analysis in verification of the proposed pathways is illustrated.

Chlorantraniliprole, an anthranilic diamide insecticide with novel mode of action, is found effective against several lepidopteran as well as coleopteran, dipteran, and hemipteran pests. The present studies were carried out to study the persistence pattern of chlorantraniliprole on sugarcane field soil following application of granule formulation. The residues of chlorantraniliprole were estimated using high performance liquid chromatograph (HPLC) and confirmed by liquid chromatograph-mass spectrometry (LC-MS/MS). Following application of chlorantraniliprole (Ferterra 0.4G) at 100 and 200 g a.i. ha(-1), the average initial deposits of chlorantraniliprole were observed to be 0.88 and 1.59 mg kg(-1), respectively. These residues dissipated below the limit of quantification (LOQ) of 0.01 mg kg(-1) after 56 days of the application of insecticides at both the dosages. The half-life values (t 1/2) of chlorantraniliprole were worked out to be 8.36 and 8.25 days, at recommended and double the recommended dosages, respectively.

This study focused on the detection and validation of the residues of the four veterinary drugs, mebendazole, clorsulon, diaveridine, and tolfenamic acid, using high performance liquid chromatography (HPLC) and an ultraviolet (UV) detector. Utilizing C18 column as a stationary phase and applying appropriate mobile phases to each analysis according to the properties of the analytes, target compounds in food samples were successfully detected and separated within 15-50 min. Additionally, in order to optimize detection, liquid-liquid extraction (LLE) and purification steps were established to minimize the endogenous peaks and their interferences. The method was validated through testing of linearity, accuracy, precision, the limit of detection (LOD) and the limit of quantification (LOQ). The LOQ levels of the four drugs were lower than the maximum residual limit, and the coefficient of determination (R(2) ) was over 0.99. The recovery results ranged from 82.3-105.2%, 79.3-83.3%, 79.4-86.0%, and 81.7-88.5% with relative standard deviations lower than 20% for mebendazole, clorsulon, diaveridine, and tolfenamic acid, respectively, corresponding to the CODEX guideline. This proposed method reduces costs and enables easier application in rural or remote areas where testing facilities or instruments often are unavailable., (Copyright © 2013 John Wiley & Sons, Ltd.)

Chlorantraniliprole (CAP) is a new anthranilic diamide insecticide acting with high efficacy on ryanodine receptors of most of the species within the Lepidoptera order and has been used worldwide. To monitor its residue in food and environmental samples, we developed an indirect competitive enzyme-linked immunosorbent assay (icELISA), based on monoclonal antibodies. The established icELISA possesses a concentration of producing 50% inhibition (IC50) of 1.60ng/mL. In addition, the assay had the average recoveries of 79-119% when CAP was fortified in tap water, soil, spinach (Spinacia oleracea), and rape (Brassica campestris). Moreover, the icELISA results of spinach samples were confirmed by high performance liquid chromatography-mass spectrometry. Overall, we demonstrate that the developed immunoassay could be used as the quick and convenient determination method of CAP residues in environmental and agricultural samples., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Background: Patients with allergic asthma have exacerbations which are frequently caused by rhinovirus infection. The antiviral tryptophan-catabolising enzyme indoleamine 2,3-dioxygenase (IDO) is induced by interferon-γ and suppressed by Th2 mediators interleukin (IL)-4 and IL-13. We hypothesised that local IDO activity after viral airway infection is lower in patients with allergic asthma than in healthy controls., Objective: To determine whether IDO activity differs between patients with allergic asthma and healthy individuals before and after rhinovirus infection., Methods: Healthy individuals and patients with allergic asthma were experimentally infected with low-dose (10 TCID50) rhinovirus 16. Blood, bronchoalveolar lavage fluid and exhaled breath condensate (for mass spectrometry by UPLC-MS/MS) were obtained before and after rhinovirus challenge., Results: IDO activity was not induced by rhinovirus infection in either group, despite increases in cold scores. However, baseline pulmonary IDO activity was lower in patients with allergic asthma than in healthy individuals. In contrast, systemic tryptophan and its catabolites were markedly higher in patients with allergic asthma. Moreover, systemic quinolinic acid and tryptophan were associated with eosinophil cationic protein (r=0.43 and r=0.78, respectively) and eosinophils (r=0.38 and r=0.58, respectively) in bronchoalveolar lavage fluid and peak asthma symptom scores after rhinovirus challenge (r=0.53 and r=0.64, respectively)., Conclusions: Rhinovirus infection by itself induces no IDO activity, but the reduced pulmonary IDO activity in patients with allergic asthma at baseline may underlie a reduced control of viral infections. Notably, the enhanced systemic catabolism of tryptophan in patients with allergic asthma was strongly related to the outcome of rhinovirus challenge in asthma and may serve as a prognostic factor.

A stability-indicating micellar liquid chromatography (MLC) method was developed and validated for the assay of floctafenine (FLF) in the presence of its degradation product and main metabolite, floctafenic acid (FLA). The analysis was carried out on a CLC Shim-Pack octyl silane (C8) column (150 x 4.6 mm id, 5 microm particle size) using a micellar mobile phase consisting of 0.15 M sodium dodecyl sulfate, 10% n-propanol, and 0.3% triethylamine in 0.02 M orthophosphoric acid (pH = 3). The mobile phase was pumped at a flow rate of 1.0 mL/min with UV detection at 360 nm. The method showed good linearity for FLF and FLA over the concentration ranges of 0.5-25.0 and 0.4-10.0 microg/mL, with LODs of 0.16 and 0.12 microg/mL, respectively. The developed method was successfully applied to the determination of FLF in commercial dispersible tablets, with mean recovery of 98.87 +/- 1.37%. Also, the proposed method was specific for the analysis of FLF in presence of the co-formulated drug thiocolchicoside in laboratory-prepared tablets, with mean recovery of 100.50 +/- 1.07%. Statistical comparison of the results obtained by the proposed MLC method with those obtained by a comparison method showed good agreement. Moreover, the method was extended to study the degradation behavior of FLF under different International Conference on Harmonization recommended conditions such as alkaline, acidic, oxidative, thermal, and photolytic. The method was further applied for direct determination of FLA as the main metabolite of FLF in human plasma without prior extraction steps, with mean recovery of 110.50 +/- 6.5%.

A method was developed for the determination of cyantraniliprole (HGW86) and its metabolite J9Z38 in watermelon and soil by ultra-performance LC (UPLC)/MS/MS. Target compounds were extracted by acetonitrile-water, cleaned up on a silica gel column, and determined by UPLC/MS/MS. Average recoveries of cyantraniliprole and J9Z38 in watermelon and soil at three levels (0.01, 0.1, and 0.5 mg/kg) ranged from 85.71 to 105.74%, with RSDs of 0.90-6.34%. The LOQs for cyantraniliprole and J9Z38 were determined to be 0.00021, 0.00015, 0.0010, and 0.00090 mg/kg in watermelon and soil samples, respectively. This method was used to determine the cyantraniliprole and J9Z38 residues in watermelon and soil samples for studies on their dissipation. The trial results showed that the half-lives of cyantraniliprole obtained after treatments were 1.1 and 4.1 days in watermelon and soil in Zhejiang, and 2.7 and 2.6 days in watermelon and soil in Hunan, respectively. The average levels of cyantraniliprole and J9Z38 residues in watermelon and soil were all < 0.01 mg/kg within the 14-day interval after treatment.

A simple and reliable analytical method was developed to detect cyantraniliprole (HGW86) and its metabolite J9Z38 in rice straw, paddy water, brown rice, and paddy soil. The fate of cyantraniliprole and its metabolite J9Z38 in rice field ecosystem was also studied. The target compounds were extracted using acetonitrile, cleaned up on silicagel or strong anion exchange column, and analyzed by ultra-performance liquid chromatography-tandem mass spectrometry. The average recoveries of cyantraniliprole and J9Z38 in rice straw, paddy water, brown rice, and paddy soil ranged from 79.0% to 108.6%, with relative standard deviations of 1.1-10.6%. The limits of quantification of cyantraniliprole and J9Z38 were 18 and 39μgkg(-1) for rice straw, 2.8 and 5.0μgkg(-1) for paddy water, 4.3 and 6.3μgkg(-1) for brown rice, and 3.9 and 5.3μgkg(-1) for paddy soil. The trial results showed that the half-lives of cyantraniliprole were 3.2, 4.4, and 6.3d in rice straw and 4.9, 2.0, and 6.2d in paddy water in Zhejiang, Hunan, and Shandong, respectively. The respective final residues of cyantraniliprole and J9Z38 in brown rice were lower than 0.05 and 0.02mgkg(-1) after 14d of pre-harvest interval. The maximum residue limit of cyantraniliprole at 0.1mgkg(-1) and dosage of 100g a.i.hm(-2), which could be considered safe to human beings and animals, were recommended., (Crown Copyright © 2013. Published by Elsevier Ltd. All rights reserved.)

Maintaining bee-friendly habitats in cities and suburbs can help conserve the vital pollination services of declining bee populations. Despite label precautions not to apply them to blooming plants, neonicotinoids and other residual systemic insecticides may be applied for preventive control of lawn insect pests when spring-flowering weeds are present. Dietary exposure to neonicotinoids adversely affects bees, but the extent of hazard from field usage is controversial. We exposed colonies of the bumble bee Bombus impatiens to turf with blooming white clover that had been treated with clothianidin, a neonicotinoid, or with chlorantraniliprole, the first anthranilic diamide labeled for use on lawns. The sprays were applied at label rate and lightly irrigated. After residues had dried, colonies were confined to forage for six days, and then moved to a non-treated rural site to openly forage and develop. Colonies exposed to clothianidin-treated weedy turf had delayed weight gain and produced no new queens whereas those exposed to chlorantraniliprole-treated plots developed normally compared with controls. Neither bumble bees nor honey bees avoided foraging on treated white clover in open plots. Nectar from clover blooms directly contaminated by spray residues contained 171±44 ppb clothianidin. Notably, neither insecticide adversely impacted bee colonies confined on the treated turf after it had been mown to remove clover blooms present at the time of treatment, and new blooms had formed. Our results validate EPA label precautionary statements not to apply neonicotinoids to blooming nectar-producing plants if bees may visit the treatment area. Whatever systemic hazard through lawn weeds they may pose appears transitory, however, and direct hazard can be mitigated by adhering to label precautions, or if blooms inadvertently are contaminated, by mowing to remove them. Chlorantraniliprole usage on lawns appears non-hazardous to bumble bees.

A field trial was conducted to study the dissipation kinetics of chlorantraniliprole 18.5 % SC on cowpea fruits. The fruit samples drawn periodically were analyzed by liquid chromatography and tandem mass detection (LC-MS/MS). The initial residues of chlorantraniliprole in cowpea fruits were found to be 0.55 mg kg(-1) which dissipated rapidly, with a half-life of 1.31 days and a waiting period of 0.62 days following first order kinetics. Processing of the fruits with solutions commonly used in households reduced the insecticide residues on cowpea fruits harvested 2 h after spraying by 47.19 %-91.70 % and 44.56 %-91.25 % on fruits plucked on the third day.

Chlorantraniliprole, an anthranilic diamide insecticide with novel mode of action is found effective against several lepidopteran as well as coleopteran, dipteran, and hemipteran pests. The present studies were carried out to study the dissipation pattern of chlorantraniliprole on cauliflower and to suggest suitable waiting period for the safety of consumers. Quick, easy, cheap, effective, rugged, and safe method was used for the extraction and cleanup of samples and the residues of chlorantraniliprole were estimated using high-performance liquid chromatograph (HPLC) and confirmed by liquid chromatograph-mass spectrometer and high-performance thin layer chromatograph. Following three applications of chlorantraniliprole (Coragen 18.5 SC) at recommended dose (9.25 g a.i. ha(-1)) and double the recommended dose (18.50 g a.i. ha(-1)), the average initial deposits of chlorantraniliprole were observed to be 0.18 and 0.29 mg kg(-1), respectively. These deposits were found to be less than the maximum residue limit of 2.0 mg kg(-1) prescribed by the Codex Alimentarius Commission. These residues dissipated below the limit of quantification of 0.10 mg kg(-1) after 3 and 5 days at recommended and double the recommended dosages, respectively. The half-life value (T (1/2)) of chlorantraniliprole was worked out to be 1.36 days following its application at recommended dosages. Hence, the use of this pesticide at recommended dosages does not seem to pose any risk, and a waiting period of 1 day is suggested for safe consumption of cauliflower curds.

A simple and efficient method based on solid-phase extraction (SPE) and high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was developed for the determination of butylene fipronil, chlorpyrifos, chlorantraniliprole, fipronil, imidacloprid, indoxacarb and phoxim residues in bamboo shoots. The sample was extracted with acetonitrile and purified by a primary secondary amine (PSA) solid-phase extraction cartridge. The detection of targets was performed by HPLC-MS/MS with multiple reaction monitoring (MRM) mode. All the seven pesticides were completely separated in 15 min with an excellent linear relationship. The limits of detection and the limits of quantification for the seven pesticides were 0.02-0.5 microg/kg and 0.08-1.5 microg/kg, respectively. The recoveries of the seven pesticides spiked in a bamboo shoot sample at the levels of 4, 8, 32 microg/kg were in the range from 76.0% to 102.6% (the RSD < or = 11.0%, n = 3). The method was successfully applied to the real bamboo shoot samples. The method has high accuracy and sensitivity, and is simple and quick. It can meet the requirement of the simultaneous determination of the seven pesticides in bamboo shoots.

The systemicity of chlorantraniliprole was investigated and evidence was collected for formulating strategies in controlling vegetable or rice pests. Systemicity of chlorantraniliprole was investigated through chlorantraniliprole application on velvetleaf (Abutilon theophrasti) leaves as well as through hydroponic experiments. The roots of velvetleaf were incubated in chlorantraniliprole solution with a concentration of 50 microg/mL; the concentrations of chlorantraniliprole in xylem above the solution parts were 3.14, 5.67, and 6.89 microg/g at 24, 48, and 72 h after treatment. When the roots were incubated in chlorantraniliprole solution with a concentration of 200 microg/mL, the concentrations of chlorantraniliprole in xylem above the solution reached 6.48, 8.76, and 10.55 microg/g at 24, 48, and 72 h after treatment. Chlorantraniliprole was not detected in the phloem above the solution after these two treatments. When chlorantraniliprole solution with a concentration of 100 microg/mL was applied on mature leaves, chlorantraniliprole was found in xylem above the mature leaves at concentrations of 0.55, 0.74, and 0.92 microg/g at 24, 48, and 72 h after the treatment. No chlorantraniliprole was detected in the leaves below the mature leaves or the phloem above them. When chlorantraniliprole solution with a concentration of 100 microg/mL was applied on apical leaves, no chlorantraniliprole was detected in the xylem or phloem below them. These results indicated that chlorantraniliprole can be transported through xylem only upward, but chlorantraniliprole has no phloem mobility in velvetleaf.

Dissipation kinetics of chlorantraniliprole was studied in sandy loam soils of sugarcane ecosystem by adopting a rapid analytical method. The recovery of chlorantraniliprole was 91.67 % when extracted with ethyl acetate as against only 65.58 % in acetonitrile-based extraction. An additional cleanup step with primary secondary amine did not enhance the recovery significantly over the no-cleanup method. The ethyl acetate-based extraction followed by direct quantification in HPLC (High-performance liquid chromatography) without any cleanup facilitated rapid quantification of chlorantraniliprole residues. The LOQ (limit of quantification) of the method was 0.01 μg/g. The half-life of chlorantraniliprole was 6.50 and 6.81 days for the recommended and double the recommended doses, respectively.

An easy, simple and efficient analytical method was standardized and validated for the estimation of residues of chlorantraniliprole in different vegetables comprising brinjal, cabbage, capsicum, cauliflower, okra, and tomato. QuEChERS method was used for the extraction and cleanup of chlorantraniliprole residues on these vegetables. Final clear extracts of ethyl acetate were concentrated under vacuum and reconstituted into high performance liquid chromatograph (HPLC) grade acetonitrile, and residues were estimated using HPLC equipped with PDA detector system, C(18) column and confirmed by liquid chromatograph mass spectrometer (LC-MS/MS), and high performance thin layer chromatograph (HPTLC). HPLC grade acetonitrile:water (80:20, v/v) was used as mobile phase @ 0.4 mL/min. Chlorantraniliprole presented distinct peak at retention time of 9.82 min. Consistent recoveries ranging from 85% to 96% for chlorantraniliprole were observed when samples were spiked at 0.10, 0.25, 0.50, and 1.00 mg/kg levels. The limit of quantification of this method was worked out to be 0.10 mg/kg., (© 2012 Institute of Food Technologists®)

A simple luminescent methodology for the simultaneous determination of mefenamic and tolfenamic acids in pharmaceutical preparations and human urine is proposed. Since the native fluorescence of both analytes is not intense, the method takes advantage of the lanthanide-sensitized luminescence, which provides a higher sensitivity. Due to the strong overlapping between the luminescence spectra of both terbium complexes, the use of luminescence decay curves to resolve mixtures of the analytes is proposed, since these curves are more selective. A factorial design with three levels per factor coupled to a central composite design was selected to obtain a calibration matrix of thirteen standards plus eight blank samples that was processed using a partial least-squares (PLS) analysis. In order to assess the goodness of the proposed method, a prediction set of synthetic samples was analyzed, obtaining recovery percentages between 90 and 104 %. Limits of detection, calculated by means of a new criterion, were 14.85 μg L(-1) and 15.89 μg L(-1) for tolfenamic and mefenamic acids, respectively. The method was tested in a pharmaceutical preparation containing mefenamic acid, obtaining recovery percentages close to 100 %. Finally, the simultaneous determination of both fenamates in human urine samples was successfully carried out by means of a correction of the above-explained model. No extraction method neither prior separation of the analytes were needed.

A rapid, simple and reliable analytical method was developed for the determination of cyantraniliprole and its major metabolite J9Z38 in pakchoi and soil by using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The sample preparation approach is known as QuEChERS, which stands for quick, easy, cheap, effective, rugged and safe. Samples were extracted with acetonitrile, and cleaned up with dispersive primary and secondary amine sorbent before analysis by UPLC-MS/MS. The limit of quantitation for cyantraniliprole and J9Z38 was 0.01 mg/kg in both pakchoi and soil. Average recoveries of cyantraniliprole and J9Z38 at three fortified levels (0.01, 0.05, 0.1 mg/kg) ranged from 77.8% to 102.5% with relative standard deviation of 1.6%-8.9%. This method has been applied to the analysis of cyantraniliprole and J9Z38 residues in real pakchoi and soil samples selected from field. The results of the residue dynamic experiment showed that the half-life of cyantraniliprole ranged from 2.9 to 6.4 days in pakchoi and 8.7 to 18.2 days in soil, respectively. The final residual levels of cyantraniliprole in pakchoi and soil from Guangdong and Shanghai were below 0.20 and 0.10 mg/kg, respectively; similarly, the final residual levels of J9Z38 in pakchoi and soil from Guangdong and Shanghai were <0.07 and 0.01 mg/kg. These results will be helpful in setting maximum residue limit guidance for cyantraniliprole in pakchoi in China.

The fate of chlorantraniliprole was studied in rice field ecosystem, and a simple and reliable analytical method was developed for determination of chlorantraniliprole in soil, rice straw, paddy water and brown rice. Chlorantraniliprole residues were extracted from samples with acetonitrile. The extract was cleaned up with QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) method, and determined by high-performance liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). The average recoveries were 76.9-82.4% from soil, 83.6-89.3% from rice straw, 95.2-103.1% from paddy water and 84.9-87.7% from brown rice. The relative standard deviation was less than 15%. The limits of detection (LODs) of chlorantraniliprole calculated as a sample concentration (S/N ratio of 3) were 0.012 μg L(-1) for paddy water, 0.15 μg kg(-1) for soil, brown rice and rice straw. The results of the kinetics study of chlorantraniliprole residue showed that chlorantraniliprole degradation in soil, water and rice straw coincided with C=0.01939e(-0.0434t), C=0.01425e(-0.8111t), and C=1.171e(-0.198t), respectively; the half-lives were about 16.0 d, 0.85 d and 3.50 d, respectively. The degradation rate of chlorantraniliprole in water was the fastest, followed by rice straw. The final residues of chlorantraniliprole on brown rice were lower than maximum residue limit (MRL) of 0.02 mg kg(-1) after 14 d Pre-Harvest Interval (PHI). Therefore, a dosage of 150 mL a.i.hm(-2) was recommended, which could be considered as safe to human beings and animals., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

A supervised field trial was conducted to study the residues of chlorantraniliprole on cabbage and cauliflower. Three applications of chlorantraniliprole at 10 days interval were made @ 9.25 and 18.50 g a.i. ha(-1). The samples of marketable size heads and curds of cabbage and cauliflower were collected at 0 and 1 day after the last application. QuEChERS sample preparation was used for the determination of chlorantraniliprole residues on cabbage heads and cauliflower curds. The residues of chlorantraniliprole were quantified by high performance liquid chromatography (HPLC) with photo diode array (PDA) detector and confirmed by high performance thin layer chromatography (HPTLC). Washing of cabbage and cauliflower with tap water removed about 17%-40% of chlorantraniliprole residues. However, boiling removed 100% of chlorantraniliprole residues on cabbage and cauliflower in both the cases.

A rapid, highly sensitive and selective method was developed for the determination of the cyantraniliprole and its major metabolite J9Z38 in cucumber, tomato and soil by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Target compounds were extracted with acetonitrile and an aliquot cleaned with primary and secondary amine. Two pairs of precursor product ion transitions for cyantraniliprole and J9Z38 were measured and evaluated. Average recoveries for cucumber, tomato and soil at three levels (10, 50 and 100 µg/kg) ranged from 74.7 to 96.2% with intra-day relative standard deviation (RSD) of 2.6-15.1% and inter-day RSD of 3.4-13.3%. The limit of quantitation for cyantraniliprole and J9Z38 were determined to be 5 and 10 µg/kg in samples (cucumber, tomato and soil), respectively. This method was used to determine the cyantraniliprole and J9Z38 residues in real cucumber, tomato and soil samples for studies on their dissipation. The trial results showed that the half-lives of cyantraniliprole obtained after treatments were 2.2, 2.8 and 9.5 days in cucumber, tomato and soil in Zhejiang, respectively, and that the average levels of cyantraniliprole and J9Z38 residues in cucumber and tomato were all <0.01 mg/kg with the interval of 10 days after treatment., (Copyright © 2011 John Wiley & Sons, Ltd.)

The main objective of this study was to understand the residue and persistence behaviour of new insecticide chlorantraniliprole in tomato fruit and soil samples. Its residue was analyzed by HPLC and it dissipated in tomato fruit and soil following first order kinetics. The results showed half life (t(1/2)) value of 3.30 and 3.66 days for chlorantraniliprole in tomato fruit and soil, respectively. According to maximum residue limit (MRL) the pre-harvest interval (PHI) of chlorantraniliprole on tomato was 8-days after the treatment.

Oats produce a group of phenolic antioxidants termed avenanthramides. These metabolites are, among food crops, unique to oats and have shown, in experimental systems, certain desirable nutritional characteristics such as inhibiting atherosclerotic plaque formation and reducing inflammation. Avenanthramides occur in both the leaves and grain of oat. In the leaves they are expressed as phytoalexins in response to crown rust (Puccina coronata) infection. The experiments reported here demonstrate that avenanthramide levels in vegetative tissue can be enhanced by treatment with benzothiadiazole (BTH), an agrochemical formulated to elicit systemic acquired resistance (SAR). The response to BTH was dramatically stronger than those produced with salicylic acid treatment. The roots of BTH treated plants also showed a smaller but distinct increase in avenanthramides. The dynamics of the root avenanthramide increase was substantially slower than that observed in the leaves, suggesting that avenanthramides might be transported from the leaves.

Ethnopharmacological Relevance: Mexican people employed infusion of leaves of Choisya ternata Kunth for their antispasmodic and "simulative properties"., Aim of the Study: In the present study the detailed GC and GC-MS analyses of the essential oil of Choisya ternata Kunth (Rutaceae) were performed. The presence of a minor constituent isopropyl N-methylanthranilate (1) was revealed among other identified volatiles. A synthesis of 1 was undertaken in order to corroborate this find and obtain gram quantities that would allow the testing of its biological activity (peripheral and central antinociceptive activity)., Materials and Methods: The oils were investigated by GC and GC-MS. Synthesized compounds were spectrally characterized (UV-Vis, IR, 1D and 2D NMR, MS). The obtained synthetic samples of compounds were assayed for peripheral and central antinociceptive activity in two models (effects on acetic acid induced writhing in mice and the hot plate test for nociception)., Results: Detailed GC and GC-MS analyses of the essential oil of Choisya ternata Kunth (Rutaceae) among 157 other identified volatiles revealed the presence of a minor constituent isopropyl N-methylanthranilate (1). Compound 1, named ternanthranin, is therefore detected as a natural product for the first time with a very restricted occurrence (samples of several citrus oils were screened for the presence of 1). The antinociceptive activities were assayed for ternanthranin, the two other synthetic analogs, methyl and propyl N-methylanthranilate, as well as the essential oil and the crude ethanol extract of the leaves. The results clearly demonstrate a very high (even significant at 0.3 mg/kg) dose dependent activity for the anthranilates (and the extracts). Isopropyl N-methylanthranilate showed the highest, while methyl N-methylanthranilate showed the lowest activity (with the methyl ester at 3 mg/kg still better than acetylsalicylic acid, at 200 mg/kg, in the first, or comparable with morphine, at 5mg/kg, in the second test)., Conclusion: This study once again revealed that detailed investigations of plant species with ethnopharmacologically documented activity may yield new natural compounds-a new alkaloid (ternanthranin), a volatile simple anthranilate that can be considered responsible for the antinociceptive activity of the crude plant extracts., (Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.)

Temporal changes in indoxacarb and chlorantraniliprole were determined in four midwestern soils by simulating commercial field applications of termiticides. Indoxacarb (0.0625 and 0.125%) and chlorantraniliprole (0.05 and 0.10%) were applied to each soil type in a rotating cement mixer to ensure uniform distribution of active ingredient (AI). Temporal and spatial changes in termiticide concentrations were determined by sampling soil cores subdivided at different depths (0-20, 20-40, and 40-61 cm) at various intervals up to 705 d after application. Percentage loss of indoxacarb was initially greater (0-180 d) than losses after 180 d. The lowest indoxacarb extractable concentrations were detected in soils closest to the surface. Chlorantraniliprole losses with time from all soils were slower than indoxacarb, with no differences observed with soil type or depth. Bioefficacy was evaluated in laboratory glass tube bioassays that monitored the distance of termite penetration into treated soils and resulting eastern subterranean termite, Reticulitermes flavipes (Kollar) (Isoptera: Rhinotermitidae), worker mortality. Bioassay data revealed that R. flavipes termites were unable to completely penetrate 50 mm of indoxacarb- and chlorantraniliprole treated soils at 0 d after treatment; however, termites were not deterred from foraging in these soils indicating no repellency to these termiticides. Termites completely penetrated (50 mm) soils treated with indoxacarb (0.0625%) by 360 d and complete penetration occurred in all soils treated with indoxacarb (0.0625 and 0.125%) by 705 d. Termites were unable to completely penetrate chlorantraniliprole-treated soils at 705 d. Mortality of termite workers was high in all chlorantraniliprole-treated soils at all sampling intervals. These data confirm that vertical differences in termiticide persistence occur in various soils.

The gas-phase dissociation reactions of chlorantraniliprole (Rynaxypyr) and cyantraniliprole (Cyazypyr) have been studied in triple-quadrupole, ion trap, and orbitrap mass spectrometers equipped with electrospray and desorption electrospray ion sources, revealing the formation of odd-electron fragment ions, the structures of which were elucidated. The odd-electron fragments were unusually abundant, and their formation is proposed to occur via a tricyclic intermediate. The applicability of the QuEChERS multiresidue method for the quantitation of chlorantraniliprole and cyantraniliprole was also assessed in this study. Four matrices representative of oily, watery, acidic, and dry crop groups were tested, with a targeted limit of quantitation (LOQ) of 0.01 mg/kg. Average recoveries ranged between 87 and 107%, with relative standard deviations (RSD) of ≤ 8%. Linear calibration functions with correlation coefficients r > 0.99 were obtained. The study provides an expansion of the QuEChERS method to include anthranilic diamides and a mass spectrometric assessment for these two novel agrochemical active ingredients.

Sensitive and accurate high-performance liquid chromatographic methods have been developed for the simultaneous determination of thiocolchicoside (TC)-glafenine (GF) (Mix I) and thiocolchicoside-floctafenine (FN) (Mix II) in their pharmaceutical formulations. The analysis for both mixtures was performed using 250 mm × 4.6 mm i.d., 5 μm particle size C18 Waters Symmetry column. The mobile phase consisted of methanol-0.035 M phosphate buffer (50:50, v/v) of pH 4.5 for Mix I and methanol-0.03 M phosphate buffer (70:30, v/v) of pH 4 for Mix II with flow rate of 1 mL/min and UV detection at 400 nm in both cases. The calibration plots were rectilinear over the concentration range of 0.2-2 μg/mL for TC in both mixtures and 20-200 μg/mL for each of GF and FN . The limits of detection for TC and GF were 0.05 μg/mL and 0.62 μg/mL, respectively, and for TC and FN were 0.02 μg/mL and 0.70 μg/mL, respectively. Additionally, the proposed methods were successfully applied to their combined tablets with average percentage recoveries of 100.35 ± 0.61 and 100.57 ± 0.72% for TC and GF respectively and for TC and FN the percentage recoveries were 101.2 ± 0.72 and 100.36 ± 0.67%, respectively. The results obtained were favorably compared with those given using the comparison methods.

An analytical method is presented for the determination of chlorantraniliprole residues in crops. Chlorantraniliprole residues were extracted from crop matrixes with acetonitrile after a water soak. The extracts were passed through a strong anion-exchange (SAX) SPE cartridge stacked on top of a reversed-phase (RP) polymer cartridge. After both cartridges were rinsed and vacuum-dried, the SAX cartridge was removed, and chlorantraniliprole was eluted from the RP polymer cartridge with acetonitrile. The acetonitrile eluate was evaporated to dryness, reconstituted, and analyzed using an LC/MS/MS instrument equipped with an atmospheric pressure chemical ionization source. The method was successfully validated at 0.010, 0.10, and 10 mg/kg for the following crop matrixes: potatoes, sugar beets (tops), lettuce, broccoli, soybeans, soybean forage, tomatoes, cucumbers, oranges, apples, pears, peaches, almonds (nutmeat), rice grain, wheat grain, wheat hay, corn stover, alfalfa forage, cottonseed, grapes, and corn grain. The average recoveries from all crop samples fortified at the method LOQ ranged from 91 to 108%, with an overall average recovery of 97%. The average recoveries from all crop samples fortified at 10 times the method LOQ ranged from 89 to 115%, with an overall average recovery of 101%. For all of the fortified control samples analyzed in this study, the overall average recovery was 99%.

The article deals with the study of two polymorphic modifications in the space groups P2(1)/c (white form) and P2(1)/n (yellow form) of the tolfenamic acid. It also describes how the white form vapor pressure temperature dependence was determined by using the transpiration method and how thermodynamic parameters of the sublimation process were calculated. We have estimated the difference between the crystal lattice energies of the two polymorphic forms by solution calorimetry and found that the crystal lattice energy of the yellow form is 6.7+/-1.2 kJ mol(-1) higher than that of the white form, whereas Gibbs free energies of the forms obtained from the vapor pressure temperature dependence are practically the same. The modifications under consideration are monotropically related. From the practical point of view, the white form is more preferable due to its lower crystal lattice energy and better performing procedure. We have also studied the solubility, solvation and transfer processes of the tolfenamic acid white form in buffers (with various values of pH and ionic strengths), n-hexane and n-octanol. The thermodynamic parameters of the investigated processes have been discussed and compared with those determined for others fenamates. In the study we estimated specific and non-specific contributions of the solvation enthalpic term of the fenamate molecules with the solvents as well. The driving forces of the transfer processes from the buffers with pH 7.4 and different ionic strengths to n-octanol were analyzed. It was found that the relationship between the enthalpic and entropic terms depends essentially on the ionic strength. For the considered fenamates the transfer processes of the neutral molecules and the ionic forms are enthalpy-determined, whereas for the niflumic acid this process is entropy-determined.

Consumers occasionally report greenish colors generated in their oat products when cooking in tap water. Here we have investigated pH and ferrous (Fe(2+)) ion as possible mechanisms for this color change. Steel-cut oat groats can turn brown-green color when cooked in alkaline conditions (pHs 9 to 12). Extraction of this color with methanol, and high-pressure liquid chromatography indicated a direct association of this color with the phenolic acid or avenanthramide content of the oat. The presence of 50 mM NaHCO(3) in water will cause oat/water mixtures to turn alkaline when cooked as CO(2) is driven off, generating OH(-) ion. Although tap water rarely, if ever, contains so much bicarbonate, bicarbonate is used as a leavening agent in baking applications. Industrial interests using baking soda or alkaline conditions during oat processing should be aware of possible off color generation. We have also found that as little as 10 ppm Fe(2+) will turn oat products gray-green when cooked. The aleurone stained darker than the starchy endosperm. Other divalent cations, such as Ca(2+) or Mg(2+) had no effect on cooked oat color. As much as 50 ppm Fe(2+) may be found in freshly pumped well water, but Fe(2+) reacts quickly with oxygen and precipitates as Fe(OH)(3). Thus, some freshly pumped well water may turn oats green when cooked, but if the water is left under atmospheric conditions for several hours, no discoloration will appear in the cooked oats.

The production of oat (Avena sativa L.) phytoalexins, avenanthramides, occurs in response to elicitor treatment with oligo-N-acetylchitooligosaccharides. In this study, avenanthramides production was investigated by techniques that provide high spatial and temporal resolution in order to clarify the process of phytoalexin production at the cellular level. The amount of avenanthramides accumulation in a single mesophyll cell was quantified by a combination of laser micro-sampling and low-diffuse nanoflow liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) techniques. Avenanthramides, NAD(P)H and chlorophyll were also visualized in elicitor-treated mesophyll cells using line-scanning fluorescence microscopy. We found that elicitor-treated mesophyll cells could be categorized into three characteristic cell phases, which occurred serially over time. Phase 0 indicated the normal cell state before metabolic or morphological change in response to elicitor, in which the cells contained abundant NAD(P)H. In phase 1, rapid NAD(P)H oxidation and marked movement of chloroplasts occurred, and this phase was the early stage of avenanthramides biosynthesis. In phase 2, avenanthramides accumulation was maximized, and chloroplasts were degraded. Avenanthramides appear to be synthesized in the chloroplast, because a fluorescence signal originating from avenanthramides was localized to the chloroplasts. Moreover, our results indicated that avenanthramides biosynthesis and the hypersensitive response (HR) occurred in identical cells. Thus, the avenanthramides production may be one of sequential events programmed in HR leading to cell death. Furthermore, the phase of the defense response was different among mesophyll cells simultaneously treated with elicitor. These results suggest that individual cells may have different susceptibility to the elicitor.

Objective: To develop a simple and rapid capillary electrophoresis (CE) method for the separation and determination of four active organic acids including salicylic acid, syringic acid, benzoic acid, and anthranilic acid in Radix Isatidis., Method: The HPCE system consisted of a fused-silica capillary column of 47.3 cm (38.3 cm to the detector) x50 microm i.d. and a mixture ofacetonitrile-borate buffer (15% acetonitrile, 25 mmol L(-1) borate, 15 mmol L(-1) beta-CD, pH 9.10) solution as the operating buffer. The applied voltage was 11.5 kV and the UV detection was set at 220 nm. The effects of the applied voltage, detection wavelength, and the pH of buffer, the concentration of buffer, acetonitrile and beta-CD were investigated., Result: The linear calibration rang was 3.0-90 mg L(-1) (r=0.9994) for salylic acid, 4.0-120 mg L(-1) (r=0.9995) for syringic acid, 2.0-60 mg L(-1) (r=0.9998) for benzoic acid and 5.0-100 mg L(-1) (r=0.9992) for anthranilic acid. The recoveries of salylic acid, syringic acid, benzoic acid and anthranilic acid were 95.9%-102.6%, 98.6%-103.4%, 98.7%-104.1%, 96.1%-104.3% respectively. The detection limits of salylic acid, syringic acid, benzoic acid and anthranilic acid were 0.7, 1.1, 1.2 and 1.5 mg L(-1), respectively.

The anthranilic and phthalic diamides, chlorantraniliprole (CAP) and flubendiamide (FLU), respectively, represent a new class of very effective insecticides that activate the ryanodine-sensitive intracellular calcium release channel (ryanodine receptor). This paper reports an analytical method for the simultaneous determination of the two insecticides on fruits and vegetables by liquid chromatography-electrospray tandem mass spectrometry operated in the positive and negative ionization switching mode. The two diamides were extracted with acetonitrile and separated on a Zorbax Column Eclipse XDB C8 (4.6 mm x 150 mm i.d., 3 microm) by isocratic elution with a mobile phase consisting of acetonitrile and water with 0.1% formic acid pumped at a flow rate of 0.4 mL/min. The diamides were selectively detected by multiple reaction monitoring for transitions of proton adduct precursor ions simultaneously: positive m/z 484.3-->285 for CAP, m/z 445.5-->169 for internal standard, and negative m/z 681.4-->253 for FLU. For CAP calibration in the positive mode was linear over a working range of 2 to 1000 microg/L with r > 0.992. The limit of detection (LOD) and limit of quantification (LOQ) for CAP were 0.8 and 1.6 microg/kg, respectively. For FLU in the negative mode the corresponding values were 1-1000 microg/L for linear working range, with r > 0.996 and 0.4 and 0.8 microg/L for LOD and LOQ, respectively. Moreover, the presence of interfering compounds in the fruit and vegetable extracts was found to be minimal. Due to the linear behavior of the MS detector response for the two analytes, it was concluded that the multiple reaction transitions of molecular ions in the ion-switching mode can be used for analytical purposes, that is, for identification and quantification of diamides in fruit and vegetable extracts at trace levels.

A rapid and new liquid chromatography-mass spectrometry with ion-trap detection method for the determination of meloxicam (MLX), flunixin meglumine (FLU), carprofen (CPF), and tolfenamic acid (TOLF) in animal tissue is described. MRLs between 10 and 500 microg kg(-1) in muscle and between 65 and 1000 microg kg(-1) in liver, from different animal species have been established in the EU for these compounds. After chemical hydrolysis, an organic extraction from homogenised tissue was performed. Final extract was injected in a liquid chromatograph with an ion-trap mass spectrometer with electrospray interface. Four identification points (one precursor and two product ions) and a minimum of one ion ratio was monitored for each compound. For quantitative purposes flunixin-D3 (FLU-D3) was used as internal standard. The method was validated using fortified blank muscle and liver from different animal species according to the 2002/657/EC European decision criteria. The decision limits (CCalpha) and detection capabilities (CCbeta) were determined and their values were at concentrations near the MRL for each substance.

Purpose: Anthranilic acid is an important, the aromatic intermediate in the degradation of tryptophan in kynurenine pathway. This compound plays an important role in the regulation of immunological processes as well shows antibacterial activity. The aim of our study was to estimate the concentration of anthranilic acid in saliva of young patients with orthodontic apparatus. We also assessed correlation between saliva anthranilic acid concentrations and time of orthodontic treatment. For the first time we have demonstrated the enhanced concentration of anthranilic acid in saliva of young orthodontic appliances., Material and Methods: The study was performed on non-stimulated, mixed saliva of patients with orthodontic appliances. The concentration of anthranilic acid and was determined by high-performance liquid chromatography., Results: The concentration of anthranilic acid was significantly higher in orthodontic patients (p = 0.043) in comparison to healthy volunteers. The mean time of orthodontic treatment was 15.0 +/- 2.03 months. We did not observe existence of correlation between anthranilic acid concentration in saliva and time of orthodontic treatment (r = -0.250; p = 0.517)., Conclusion: These results might indicate that anthranilic acid can be one of many factors initiating of periodontal disease in orthodontic appliances.

Purpose: It has been showed that tryptophan (TRP) degradation has been linked to modulation of cancer cell proliferation. The aim of our study was to estimate the concentration of TRP and its derivatives, such as anthranilic (AA) and kynurenic acid (KYNA) in plasma, saliva, squamous cell carcinoma (SCC) tissues and healthy oral mucosa in patients with oral SCC., Material and Methods: The study was performed on plasma, non-stimulated, mixed saliva and squamous cell carcinoma tissues and healthy oral mucosa in patients with oral SCC. The concentration of TRP and its metabolites were determined by high-performance liquid chromatography (HPLC)., Results: In plasma the concentration of TRP was 33.73 +/- 2.52 microM, of KYNA was 26.97 +/- 5.35 nM and of AA was 32.40 +/- 2.30 nM. In saliva the concentration of TRP was 3.81 +/- 0.62 microM, of KYNA was 8.06 +/- 1.86 nM and of AA was 20.41 +/- 10.77 nM. In cancer tissues the levels of TRP (30.21 +/- 5.88 microM), KYNA (15.85 +/- 1 .82 nM) and AA (265.32 +/- 1 51.45 nM) were higher in respect to the concentration of TRP (13.28 +/- 0.62 microM), KYNA (12.75 +/- 2.28 nM) and AA (31.68 +/- 8.89 nM) in normal tissues. The increase in the content of TRP, KYNA and AA in cancer tissues reached 127.48 +/- 5.95%, 24.31 +/- 4.35% and 737.50 +/- 206.96%, respectively., Conclusions: Our study has demonstrated the change of TPR metabolism, which is reflected by the increase TRP, AA and KYNA concentrations in patients with oral squamous cell carcinoma. We can suppose that these substances may be one of many factors responsible for cancer development.

Infrared spectroscopy has been used to characterize the polymorphic purity as well as to study the thermal conversion of three of the more common fenamates between their different crystalline forms via measuring changes in the NH stretch region that occur between 3300 and 3350 wavenumbers. Shifts in band frequency for mefenamic acid result from differences in internal hydrogen bonding between the NH group and either the carbonyl or hydroxyl groups of the acid moiety. Due to out-of-plane rotations about the central N-C(ring2) bond additional polymorphic states have been suggested for flufenamic and tolfenamic acids. Rates of conversion are given for flufenamic, mefenamic, and tolfenamic acids at temperatures between 85 and 160 degrees C depending on the polymorphic transition for a particular analyte. Subsequently, these rates are used to calculate the activation energy for the observed polymorphic transition. Values of 71.6, 49.0, and 50.8 kcal/mol are obtained respectively for (1) the polymorph I to II transition of mefenamic acid, (2) the polymorph I to II transition of tolfenamic acid, and (3) the polymorph III to I transition of flufenamic acid.

Five polymorphic forms of tranilast were characterized by thermal, diffractometric, and spectroscopic techniques. The crystal structures of the most stable anhydrous form (Form I), a chloroform solvate, and a dichloromethane solvate were determined from single-crystal X-ray analysis. Two additional anhydrous forms of tranilast (Forms II and III) were also studied, but were not amenable to SCXRD. All five forms were also analyzed using solid-state nuclear magnetic resonance, Fourier transform infrared, and Fourier transform-Raman spectroscopy, and thermal methods. From the trends observed in the crystal structures and the spectral data, some conclusions can be made about hydrogen bonding, molecular conformation, and crystal packing differences in the polymorphs and solvates. Form II was found to be a spectroscopically distinctive polymorph that is probably missing an important intramolecular hydrogen bond coupled with a conformational change. In contrast, Form III was found to be more similar to the crystallographically characterized forms, and is more likely a packing and hydrogen-bonding polymorph with a weakened intermolecular hydrogen-bonding interaction relative to the other forms. From a pharmaceutical development perspective, it is shown that although the anhydrous forms of tranilast have similar thermal properties, they can be reliably distinguished by spectroscopic methods., (Copyright 2005 Wiley-Liss, Inc. and the American Pharmacists Association.)

Two fundamentally different mechanisms of ligand binding are commonly encountered in biological kinetics. One mechanism is a sequential multistep reaction in which the bimolecular binding step is followed by first-order steps. The other mechanism includes the conformational transition of the macromolecule, before the ligand binding, followed by the ligand binding process to one of the conformational states. In stopped-flow kinetic studies, the reaction mechanism is established by examining the behavior of relaxation times and amplitudes as a function of the reactant concentrations. A major diagnostic tool for detecting the presence of a conformational equilibrium of the macromolecule, before the ligand binding, is the decreasing value of one of the reciprocal relaxation times with the increasing [ligand]. The sequential mechanism cannot generate this behavior for any of the relaxation times. Such dependence is intuitively understood on the basis of approximate expressions for the relaxation times that can be comprehensively derived, using the characteristic equation of the coefficient matrix and polynomial theory. Generally, however, the used approximations may not be fulfilled. On the other hand, the two kinetic mechanisms can always be distinguished, using the approach based on the combined application of pseudo-first-order conditions, with respect to the ligand and the macromolecule. The two experimental conditions differ profoundly in the extent of the effect of the ligand on the protein conformational equilibrium. In a large excess of the ligand, the conformational equilibrium of the macromolecule, before the ligand binding, is strongly affected by the binding process. However, in a large excess of the macromolecule, ligand binding does not perturb the internal equilibrium of the macromolecule. As a result, the normal mode, affected by the conformational transition, is absent in the observed relaxation process. In the case of a sequential mechanism, the number of relaxation times is not altered by different pseudo-first-order conditions. Thus, the approach provides a strong diagnostic criterion for detecting the presence of the conformational transition of the macromolecule and establishing the correct mechanism. Application of this approach is illustrated for the binding of 3'-O-(N-methylantraniloyl)-5'-diphosphate to the E. coli DnaC protein.

Oligosaccharides were derivatized by reductive amination using 2-aminobenzamide (2-AB) and analyzed by matrix-assisted laser desorption/ionization two-stage time-of-flight (MALDI-TOF/TOF) tandem mass spectrometry (MS/MS) in the positive ion mode. The major signals were obtained under these conditions from the [M+Na]+ ions for all 2-AB-derivatized oligosaccharides. A systematic study was conducted on a series of 2-AB-derivatized oligosaccharides to allow rationalization of the fragmentation processes. The MALDI-TOF/TOF-MS/MS spectra of the [M+Na]+ ions of 2-AB-derivatized oligosaccharides were dominated by glycosidic cleavages. These fragments originating both from the reducing and the non-reducing ends of the oligosaccharide yield information on sequence and branching. Moreover, the MALDI-TOF/TOF-MS/MS spectra were also characterized by abundant cross-ring fragments which are very informative on the linkages of the monosaccharide residues constituting these oligosaccharides. MALDI-TOF/TOF-MS/MS analysis of 2-AB-derivatized oligosaccharides, by providing structural information at the low-picomole level, appears to be a powerful tool for carbohydrate structural analysis., (Copyright (c) 2005 John Wiley & Sons, Ltd.)