Dissipation of Herbicide Methiozolin and Its Metabolites in Aerobic Sediment-Water Systems.

Methiozolin is a novel herbicide for controlling annual bluegrass. After applying 14C labelled methiozolin in two sediment (clay loam and sand)-water systems under aerobic conditions, its distribution, half-life, and metabolites within 300 days were investigated. The mass balance ranged within 92.0%-104.4% of applied radioactivity (AR). Radioactivity in the water declined sharply from 94.4% to 0.5% AR, while in the sediment it increased to 83.9% AR at 14 days before declining to 9.1% AR. The volatiles were minimal (< 0.5% AR), and the evolved labelled CO2 accounted for up to ~ 33.4% AR. From Radio-HPLC analysis, labelled methiozolin in water decreased from 108.9% to 0% AR, while a maximum of 15.1% AR remained in the sediment at the end. Eight metabolites were detected, all at minor levels and accounting for < 5.5% AR. The half-life of labelled methiozolin in the total sediment-water systems were 50.7 and 38.7 days for clay loam and sand, respectively.

Anaerobic Degradation of [14C]Methiozolin under Aquatic Sediment Conditions.

The fate of methiozolin under anaerobic conditions was investigated in clay loam with a high organic carbon content and sandy loam with a low carbon content using [dihydroisoxazole ring-14C] and [phenyl-14C] radiolabels. The sediment/water ratio was 1:3 based on the dry weight:volume (w/v) ratio; the incubations lasted up to 355 days after the treatment (DAT) and were performed in the dark at 20.4 ± 0.7 °C. The overlying water flow-through systems consisted of glass vessels containing sediment with traps for [14C]carbon dioxide and [14C]volatiles. The samples were collected and analyzed at 0, 3, 7, 14, 50, 100, 200, and 355 DAT. The water and sediment samples were extracted with solvent systems, centrifuged, concentrated, and analyzed by liquid scintillation counting and a high-performance liquid chromatography (HPLC) system equipped with a flow scintillation analyzer. Following extraction, the sediments were air-dried, and the subsamples were combusted. [14C]Methiozolin was degraded in the water phase and partitioned rapidly into the sediments, where it was further degraded to other metabolites, which were identified by HPLC and liquid chromatography- or gas chromatography-tandem mass spectrometry (MS/MS) with authentic standards. The dissipation of methiozolin from the overlying water was rapid (with half-lives of 1.1-1.8 and 3.6-4.9 days in the clay loam and sandy loam, respectively). However, methiozolin dissipation from the sediment phase and the whole system was much slower than from the water phase (with half-lives of 122.0-220.0 and 110.0-130.0 days in the sediment phase of the clay loam and sandy loam and 116.0-166.0 and 70.8-85.7 days in the whole system of the clay loam and sandy loam, respectively).

Developmental toxicity assessment of the new turf herbicide, methiozolin ([5-(2,6-difluorobenzyl)oxymethyl-5-methyl-3,3(3-methylthiophen-2-yl)-1,2-isoxazoline]), in rabbits.

Methiozolin is a new herbicide to control annual bluegrass (Poa annua L.) and large crabgrass (Digitaria sanguinalis (L.) Scop.) in various turfgrasses. The potential of methiozolin to induce maternal and developmental toxicity was investigated in the pregnant New Zealand White Rabbits. Methiozolin was, at dose levels of 0, 125, 250 and 500 mg/kg/day, administered by oral gavage to artificially inseminated rabbits (25 females per group) from days 6 to 28 of gestation. All does were subjected to Cesarean section on day 29 of gestation. At 500 mg/kg/day, treatment-related toxicities including abortion (10/22), decreased mean body weight, weight gain, net body weight change, reduced food consumption and decreased fetal weight were observed. At 125 and 250 mg/kg/day, no signs of maternal and developmental toxicity were observed. There were no treatment-related external, visceral and skeletal abnormalities of fetuses at all doses tested. In the current experimental conditions, the no observed adverse effect levels (NOAELs) of methiozolin are considered to be 250 mg/kg/day for does and prenatal development.

Adenine suppresses IgE-mediated mast cell activation.

Nucleobase adenine is produced by dividing human lymphoblasts mainly from polyamine synthesis and inhibits immunological functions of lymphocytes. We investigated the anti-allergic effect of adenine on IgE-mediated mast cell activation in vitro and passive cutaneous anaphylaxis (PCA) in mice. Intraperitoneal injection of adenine to IgE-sensitized mice attenuated IgE-mediated PCA reaction in a dose dependent manner, resulting in a median effective concentration of 4.21 mg/kg. In mast cell cultures, only adenine among cytosine, adenine, adenosine, ADP and ATP dose-dependently suppressed FcɛRI (a high affinity receptor for IgE)-mediated degranulation with a median inhibitory concentration of 1.6mM. It also blocked the production of LTB4, an inflammatory lipid mediator, and inflammatory cytokines TNF-α and IL-4. In addition, adenine blocked thapsigargin-induced degranulation which is FcɛRI-independent but shares FcɛRI-dependent signaling events. Adenine inhibited the phosphorylation of signaling molecules important to FcɛRI-mediated allergic reactions such as Syk, PLCγ2, Gab2, Akt, and mitogen activated protein kinases ERK and JNK. From this result, we report for the first time that adenine inhibits PCA in mice and allergic reaction by inhibiting FcɛRI-mediated signaling events in mast cells. Therefore, adenine may be useful for the treatment of mast cell-mediated allergic diseases. Also, the upregulation of adenine production may provide another mechanism for suppressing mast cell activity especially at inflammatory sites.

Methiozolin [5-(2,6-difluorobenzyl)oxymethyl-5-methyl-3,3(3-methylthiophen-2-yl)-1,2-isoxazoline], a new annual bluegrass (Poa annua L.) herbicide for turfgrasses.

BACKGROUND: Selective control of annual bluegrass (Poa annual L.) has been difficult in turfgrasses. The potential of methiozolin in this area was investigated. RESULTS: Methiozolin was safe on established zoysiagrass (Zoysia japonica Steud.), creeping bentgrass (Agrostis palustris Huds.), Kentucky bluegrass (Poa pratensis L.), and perennial ryegrass (Lolium perenne L.) at 1000 g ha(-1) , and controlled annual bluegrass with GR50 values of 23, 52, 104, and 218 g ha(-1) at PRE, two-, four- and eight-leaf stage, respectively, in the greenhouse. When applied at early flowering, methiozolin suppressed >80% of annual bluegrass seed heads at 2000 g ha(-1) . (14) C-Methiozolin was readily absorbed by both leaves and roots, but translocation was mainly acropetal. No herbicidal activity resulted from application to the leaf only; however, application to the soil surface only showed equivalent herbicidal activity to that of broadcast application to the leaf and soil. Methiozolin at 500 to 1000 g ha(-1) provided 80 to 100% control of annual bluegrass when applied in the fall with acceptable and temporary injury to creeping bentgrass, and about 60% control when applied in the spring with no bentgrass injury in the field. CONCLUSION: Methiozolin is an excellent candidate for annual bluegrass management in turfgrasses.

In vivo absorption, distribution, excretion, and metabolism of a new herbicide, methiozolin, in rats following oral administration.

Methiozolin [5-(2,6-difluorobenzyl)oxymethyl-5-methyl-3-(3-methylthiophen-2-yl)-1,2-isoxazoline] is a new turf herbicide that controls annual bluegrass in various cool- and warm-season turfgrasses. The present study is the first report elucidating absorption, tissue distribution, excretion, and metabolism of methiozolin in rats. The pharmacokinetic parameters in the blood were observed as follows: t(max) = 6 h, C(max) = 168.7 µg equiv/mL, t(½) = 49.4 h, AUC120 = 9921.5 µg equiv·h/mL, and clearance = 39.2 mL/h/kg. Those parameters and the depletion curve for ¹4C in the plasma were very similar to those in the blood. Total excretion through urine and feces was 24.3 and 68.9%, respectively, during 120 h after administration; however, there was no excretion through expired air. The radioactivity excreted through bile was 40.1% of that administered. Excreted radioactivity peaked between 24 and 48 h, showing 51.0% of total excretion within 48 h. The orally administered ¹4C distributed across various tissues within 12 h after administration, showing 14.0% of the dosed, and was eliminated from all tissues without accumulation. Numerous minor metabolites (<4% of the dosed) in urine and fecal extract were detected within 72 h, and two of those were identified. The identified metabolites were Met-1 (glucuronic acid conjugate), 6-[5-(5-((2,6-difluorobenzyloxy)methyl)-4,5-dihydro-5-methylisoxazol-3-yl)-4-methylthiophen-2-yloxy]-tetrahydro-3,4,5-trihydroxy-2H-pyran-2-carboxylic acid, and Met-2, [2-(5-((2,6-difluorobenzyloxy)methyl)-4,5-dihydro-5-methylisoxazol-3-yl)thiophen-3-yl]methanol. Conclusively, methiozolin was shown to be readily absorbed in the gastrointestinal tract, distributed throughout the tissues within 12 h, metabolized extensively, and eliminated through urine and feces mostly within 48 h, without tissue accumulation.

Soil metabolism of [14C]methiozolin under aerobic and anaerobic flooded conditions.

Methiozolin is a new turf herbicide controlling annual bluegrass in various cool- and warm-season turfgrasses. This study was conducted to investigate the fate of methiozolin in soil under aerobic and anaerobic flooded conditions using two radiolabeled tracers, [benzyl-(14)C]- and [isoxazole-(14)C]methiozolin. The mass balance of applied radioactivity ranged from 91.7 to 104.5% in both soil conditions. In the soil under the aerobic condition, [(14)C]methiozolin degraded with time to remain by 17.9 and 15.9% of the applied in soil at 120 days after treatment (DAT). [(14)C]Carbon dioxide and the nonextractable radioactivity increased as the soil aged to reach up to 41.5 and 35.7% for [benzyl-(14)C]methiozolin at 120 DAT, respectively, but 36.1 and 39.8% for [isoxazole-(14)C]methiozolin, respectively, during the same period. The nonextractable residue was associated more with humin and fulvic acid fractions under the aerobic condition. No significant volatile products or metabolites were detected during this study. The half-life of [(14)C]methiozolin was approximately 49 days in the soil under the aerobic condition; however, it could not be estimated in the soil under the anaerobic flooded condition because [(14)C]methiozolin degradation was limited. On the basis of these results, methiozolin is considered to undergo fast degradation by aerobic microbes, but not by anaerobic microbes in soil.

Metabolism of a new herbicide, [(14)c]pyribenzoxim, in rice.

The in vivo metabolism of a new herbicide pyribenzoxim (benzophenone Ο-[2,6-bis(4,6-dimethoxypyrimidin-2-yloxy)benzoyl]oxime) in rice was carried out using container trials. Two radiolabeled forms of [carbonyl-(14)C]pyribenzoxim (P1) and [ring-(14)C(U)]pyribenzoxim (P2) were treated separately as formulations for foliar treatment by single applications of 50 g of active ingredient (ai)/ha at the 4-6 leaves stage. At 0, 7, 30, and 60 days after treatment (DAT), samples of panicle, foliage/rest of plant, and roots were taken for analysis. Upon harvest (120 DAT), rice plants were separated into grain, husk, straw, and root parts. Total radioactive residues (TRRs) at each sampling date were determined to show that the final radioactive residues at harvest were low in grain, husk, straw, and roots, accounting for <17 ppb. The concentration of final residues in the rice plant decreased rapidly, and less than 0.1% of initial TRRs remained at harvest. At 7 DAT, metabolite 1 [M1, 2,6-bis(4,6-dimethoxypyrimidin-2-yloxy)benzoic acid] and two unknown compounds (other-1 and other-2) were detected in foliage extract, accounting for 3.5% TRRs (21.0 ppb), 3.1% TRRs (19.0 ppb), and 9.0% TRRs (54.3 ppb), respectively, while 26.1% of M1 was observed in solvent wash. Any other metabolites were not detected in the plant, including expected metabolite M3 (benzophenone oxime). On the basis of the results obtained, a metabolic pathway of pyribenzoxim in a rice plant was proposed.

Soil metabolism of a new herbicide, [14C]Pyribenzoxim, under flooded conditions.

To elucidate the fate of a new pyrimidinyloxybenzoic herbicide, pyribenzoxim, a soil metabolism study was carried out with [14C]pyribenzoxim applied to a sandy loam soil under flooded conditions. The material balance of applied radioactivity ranged from 96.4 to 104.4% and from 96.1 to 101.9% for nonsterile and sterile soils, respectively. The half-life of [14C]pyribenzoxim was calculated to be approximately 1.3 and 9.4 days for nonsterile and sterile soils, respectively. The metabolites identified during the study were 2,6-bis(4,6-dimethoxypyrimidin-2-yloxy)benzoic acid (M1) and 2-hydroxy-6-(4,6-dimethoxypyrimidin-2-yloxy)benzoic acid (M2), resulting from the cleavage of the ester bond and subsequent hydrolysis. The nonextractable radioactivity levels increased to 37.8% for nonsterile conditions at 50 days after treatment and to 38.2% for sterile conditions at 60 days after treatment. Fractionation of the nonextractable soil residues indicated that bound radioactivity was associated mainly with humin fraction. No significant volatile products or [14C]carbon dioxide was observed during the study. On the basis of these results, pyribenzoxim is considered to undergo rapid degradation in soil by microbial and chemical reactions, mainly hydrolysis, which limits its transfer to and accumulation in lower soil layers and groundwater. Therefore, the possibility of environmental contamination from the use of pyribenzoxim is expected to be very low.

Identification of rat urinary and fecal metabolites of a new herbicide, pyribenzoxim.

Rats were treated with pyribenzoxim (O-[2,6-bis[(4,6-dimethoxy-2-pyrimidinyl)oxy]benzoyl]oxime), a new herbicide, to investigate the related metabolites in urine and feces. Metabolites were identified using LC/MS (electrospray ionization) and GC/MS (electron impact ionization) following the relatively simple and rapid extraction and purification procedures. Three metabolites were identified in urine either from oral gavage or intravenous (iv) injection. They were benzophenone oxime (BO), benzophenone oxime glucuronide (BOG), and 2-hydroxy-6-(4,6-dimethoxypyrimidin-2-yloxy)benzoic acid (HDB). Benzophenone oxime was present in larger quantity than BOG and HDB in urine from oral treatment, while the case was opposite in urine from iv treatment. Glucuronide conjugate was confirmed unambiguously by enzyme hydrolysis. 2,6-Bis(4,6-dimethoxypyrimidin-2-yloxy)benzoic acid (KIH-2023) and benzophenone were identified in feces. Benzophenone was confirmed by GC/MS and HPLC/DAD since LC/MS could not produce an ESI spectrum. On the basis of the results obtained, a metabolic map of pyribenzoxim is proposed.

Degradation of the sulfonylurea herbicide LGC-42153 in flooded soil.

LGC-42153, 2-fluoro-1-[3-(4,6-dimethoxypyrimidin-2-ylcarbamoylsulfamoyl)pyridin-2-yl]propyl methoxyacetate, is a new sulfonylurea herbicide for use in rice. Its breakdown and metabolism were studied in soil under flooded condition using radioactive tracers labelled at either the propyl group or the pyrimidine ring. The half-life of LGC-42153 was approximately 3.0 days. The mass balance over 120 days ranged from 94.0 to 104.2% of applied radiocarbon, and no significant amount of volatiles or [14C]carbon dioxide were observed. Solvent non-extractable radiocarbon reached 11 approximately 14% of applied radiocarbon at 120 days after treatment. The major metabolic reaction was the cleavage of the carboxyl ester bond to give 1-(4,6-dimethoxypyrimidin-2-yl)-3-[2-(1-hydroxy-2-fluoropropyl)pyridine-3-sulfonyl]urea, which underwent hydrolysis of the sulfonylurea bridge giving 2-(1-hydroxy-2-fluoro)propyl-3-pyridinesulfonamide and 4,6-dimethoxy-2-aminopyrimidine.

Degradation of the sulfonylurea herbicide LGC-42153 in flooded soil.

LGC-42153, 2-fluoro-1-[3-(4,6-dimethoxypyrimidin-2-ylcarbamoylsulfamoyl)pyridin-2-yl] propyl methoxyacetate, is a new sulfonylurea herbicide for use in rice. Its breakdown and metabolism was studied in soil under flooded conditions using two radioactive tracer compounds labelled at either the propyl group or the pyrimidine ring. The half-life of LGC-42153 was approximately 3.0 days. The mass balance over 120 days ranged from 94.0 to 104.2% of applied radiocarbon, and no significant amount of volatiles or [14C]carbon dioxide were observed. Solvent non-extractable radiocarbon reached about 11-14% of applied radiocarbon at 120 days after treatment. The major metabolic reaction was the cleavage of the carboxyl ester bond to give 1-(4,6-dimethoxypyrimidin-2-yl)-3-[2-(1-hydroxy-2-fluoropropyl)pyridine-3-sulfonyl]urea, which underwent hydrolysis of the sulfonylurea bridge giving 2-(1-hydroxy-2-fluoro)propyl-3-pyridinesulfonamide and 4,6-dimethoxy-2-aminopyrimidine.

Aerobic soil metabolism of a new herbicide, LGC-42153.

To elucidate the fate of a new sulfonylurea herbicide, LGC-42153 [N-((4,6-dimethoxypyrimidin-2-yl)aminocarbonyl)-2-(1-methoxyacetoxy-2-fluoropropyl)-3-pyridinesulfonamide], in soil, an aerobic soil metabolism study was carried out for 120 days with [(14)C]LGC-42153 applied to a loamy soil. The material balance ranged from 90.7 to 101.5% of applied herbicide. The half-life of [(14)C]LGC-42153 was calculated to be approximately 9.0 days. The degradation products resulted from the cleavage of the sulfonylurea bridge. The metabolites identified during the study were N-((4,6-dimethoxypyrimidin-2-yl)aminocarbonyl)-2-(1-hydroxy-2-fluoropropyl)-3-pyridinesulfonamide, 2-(1-hydroxy-2-fluoropropyl)-3-pyridinesulfonamide, and 4,6-dimethoxy-2-aminopyrimidine. No significant volatile products or [(14)C]carbon dioxide was observed during the study. Nonextractable (14)C-residue reached 14.4-30.5% of applied material at 120 days after treatment, and radioactivity was distributed mostly in the humin and fulvic acid fractions.