Stereoisomeric separation of the chiral herbicide profoxydim and residue method development in rice by QuEChERS and LC-MS/MS.

A method for the simultaneous determination of the four stereoisomers of the chiral herbicide profoxydim in rice and husk was developed using the QuEChERS method and LC-tandem mass spectrometry. Four polysaccharide-based chiral stationary phase columns were evaluated. All four stereoisomers were successfully separated on a Chiracel OJ-3R column. The effects of mobile phase, modifiers, mobile phase flow rate and temperature on the separation were also investigated. Different QuEChERS methods were compared for the development of an optimized sample preparation procedure. The method, following SANTE guidelines, showed excellent linearity (R2 ≥ 0.99), the LODs were below 4.0 µg kg-1, and the LOQs did not exceed 12.5 µg kg-1. The overall average recoveries at three levels (12.5, 25.0 and 250 µg kg-1) ranged from 76.77 % to 106.53 %, with RSD values less than 7 %. The method is demonstrated to be convenient and reliable for the routine monitoring of profoxydim stereoisomers in rice and husk.

Assessing the effect of organic amendments on the degradation of profoxydim in paddy soils: Kinetic modeling and identification of degradation products.

The fate and behavior of herbicides can be altered in an unpredictable way when organic amendments are added to soil as a beneficial management tool. The objective of this work was to investigate the effect exerted by the addition of two different organic amendments (alperujo compost and biochar) to soil in the degradation of one of the most relevant new generation rice herbicides, profoxydim. In unamended soils, the degradation of profoxydim was quite fast and was governed by both chemical (DT50steril soil = from 1.52 to 9.21 days) and microbial (DT50nonsterile soil = from 0.47 to 0.53 days) processes. Alperujo- and biochar-amended soils significantly increased the persistence of the herbicide in both soils, especially in the presence of biochar, due to the high capacity absorption of this amendment, increasing DT90 from 1.92 to 3.54 days for DT90unamended to 41.02-48.41 days for DT90biochar amended. Different kinetics models applied to fit the observed dissipation datasets showed that a HS biphasic model fits well with the dissipation of profoxydim in amended and unamended soils. For the first time, five degradation products (DPs) were identified by HPLC-QTOF-MS/MS in soil and a degradation pathway was described. Main DP was generated via oxidation of the sulfur atom to give rise to the corresponding sulfoxide derivative, with this DP being more persistent than the active substance. These outcomes can be very useful for the assessment of the environmental risk associated with the use of profoxydim in rice crops and the application of organic amendments as potential measures for minimizing the risk of contamination of natural water resources.

QSAR/QSPR models based on quantum chemistry for risk assessment of pesticides according to current European legislation.

In Europe, agencies and official organizations involved in the pesticide control such as the EFSA, ECHA, JRC and ECETOC or even the OECD are pointing out that the software tools based on quantitative structure relationship models, i.e. QSAR and QSPR, have a huge potential to improve the pesticide risk assessment process. In this sense, these non-animal test methods can promote the competitiveness of agriculture in this region: the consumer safety is increased with them due to the possibility of perform an overall better risk assessment of the degradation products and metabolites from pesticides. However, the use of theses computational-based (in silico) tools must be much more systematised and harmonised, improving their validation and including case studies to test them. To open databases, incorporating critical data in an orderly manner for building the models, becomes also necessary. Moreover, quantum chemistry through the Density Functional Theory should be promoted as tool for calculation of quantum descriptors, especially for the study of similar compounds with the same carbon skeleton but differing substitution patterns, e.g. isomers.

Optimal growth of Dunaliella primolecta in axenic conditions to assay herbicides.

To develop an assay for herbicides in marine environments using microalgae, we have optimized the specie, cell culture media and physical conditions to obtain maximal cellular densities in a 96 well micro format to allow mass assays. We first surveyed several species of 7 unicellular eukaryotic algae genera (Dunaliella, Tetraselmis, Chlorella, Ellipsoidon, Isochrysis, Nannochloropsis, and Phaeodactylum) for vigorous in vitro axenic growth. Once the genus Dunaliella was selected, Dunaliella primolecta was preferred among 9 species (bioculata, minuta, parva, peircei, polymorpha, primolecta, quartolecta, salina and tertiolecta) because it showed the highest growth rates. The components (oligo elements, sugars, amino acids and vitamins) and conditions (light, CO(2), temperature) of the culture media were further optimized to obtain the highest cellular densities (up to 60x10(6)cellsml(-1)) and the shortest cell cycle duration ( approximately 12h) for D. primolecta. Then the toxicity of four representative herbicides, alloxydim, and sethoxydim (inhibitors of acetyl-coA carboxilase), metamitron (inhibitor of photosynthesis) and clopyralid (inhibitor of respiration), were assayed on the optimal culture conditions for D. primolecta during 96h. The results showed that D. primolecta was susceptible to those herbicides in the following order: metamitron > sethoxydim > alloxydim. In contrast, clopyralid did not have any effects. Therefore, D. primolecta microcultures can be used to assay a large number of samples for the presence of herbicides under a saline environment.