Jacques Einhorn

Flavonoid diversity and biosynthesis in seed of Arabidopsis thaliana

Functional characterization of genes involved in the flavonoid metabolism and its regulation requires in-depth analysis of flavonoid structure and composition of seed from the model plant Arabidopsis thaliana. Here, we report an analysis of the diverse and specific flavonoids that accumulate during seed development and maturation in wild types and mutants. Wild type seed contained more than 26 different flavonoids belonging to flavonols (mono and diglycosylated quercetin, kaempferol and isorhamnetin derivatives) and flavan-3-ols (epicatechin monomers and soluble procyanidin polymers with degrees of polymerization up to 9). Most of them are described for the first time in Arabidopsis. Interestingly, a novel group of four biflavonols that are dimers of quercetin-rhamnoside was also detected. Quercetin-3-O-rhamnoside (the major flavonoid), biflavonols, epicatechin and procyanidins accumulated in the seed coat in contrast to diglycosylated flavonols that were essentially observed in the embryo. Epicatechin, procyanidins and an additional quercetin-rhamnoside-hexoside derivative were synthesized in large quantities during seed development, whereas quercetin-3-O-rhamnoside displayed two peaks of accumulation. Finally, 11 mutants affected in known structural or regulatory functions of the pathway and their three corresponding wild types were also studied. Flavonoid profiles of the mutants were consistent with previous predictions based on genetic and molecular data. In addition, they also revealed the presence of new products in seed and underlined the plasticity of this metabolic pathway in the mutants.

Profiling of phenolic glycosidic conjugates in leaves of Arabidopsis thaliana using LC/MS

Profiling of plant secondary metabolites is still a very difficult task. Liquid chromatography (LC) or capillary electrophoresis hyphenated with different kinds of detectors are methods of choice for analysis of polar, thermo labile compounds with high molecular masses. We demonstrate the applicability of LC combined with UV diode array or/and mass spectrometric detectors for the unambiguous identification and quantification of flavonoid conjugates isolated from Arabidopsis thaliana leaves of different genotypes and grown in different environmental conditions. During LC/UV/MS/MS analyses we were able to identify tetra-, tri-, and di-glycosides of kaempferol, quercetin and isorhamnetin. Based on our results we can conclude that due to the co-elution of different chemical compounds in reversed phase HPLC systems the application of UV detectors does not allow to precisely profile all flavonoid conjugates existing in A. thaliana genotypes. Using MS detection it was possible to unambiguously recognize the glycosylation patterns of the aglycones. However, from the mass spectra we could not conclude neither the anomeric form of the C-1 carbon atoms of sugar moieties in glycosidic bonds between sugars or sugar and aglycone nor the position of the second carbon involved in disaccharides. The applicability of collision induced dissociation techniques (CID MS/MS) for structural analyses of the studied group of plant secondary metabolites with two types of analyzers (triple quadrupole or ion trap) was demonstrated.

Natural sunlight NO(3)(-)/NO(2)(-)-induced photo-degradation of phenylurea herbicides in water.

The nitrate-induced photodegradation of phenylureas in water was demonstrated to occur efficiently using natural sunlight irradiation. The kinetics of disappearance was found to be dependent on the inducer and substrate concentrations, the phenylurea structure and the origin and composition of the aqueous matrix including the presence of nitrite. The measured effects under sunlight were of the same order of those measured previously in the lab using our solar light simulated system. However, by-product distribution might differ substantially particularly considering the nitration pathway.

Detection of isoflavonoids and their glycosides by liquid chromatography/electrospray ionization mass spectrometry in root extracts of lupin (Lupinus albus)

HPLC combined with electrospray ionization MS (LC–ESI-MS) was used for the identification of isoflavonoids and their glycosides in extracts from white lupin (Lupinus albus) tissues. The applicability of LC–ESI-MS for the simultaneous analysis of secondary metabolites of different polarities (free aglycones and their glycosides) is demonstrated. For optimization of the chromatographic and MS conditions, genistin (genistein 7-O-β-glucoside) and rutin [quercetin 3-O-β-(6″-O-α-rhamnosyl glucoside)] were used as standards. The detection limit for the analysis of genistin by HPLC was determined to be 20u2005ng (ca. 50u2005pmol). The analytical conditions established were employed to analyse isoflavonoids and their glycosidic conjugates obtained from lupin root extracts by solid phase extraction. Four glycosides and seven isoflavonoid aglycones were detected in the analysed samples. Copyright

Identification of the sex pheromone of the maritime pine scale Matsucoccus feytaudi.

(2E,4E)-4,6,10-trimethyl-2,4-dodecadien-7-one and (2E,4Z)-4,6,10-trimethyl-2,4-dodecadien-7-one have been identified mostly through 2D NMR and MS/MS techniques as primary and secondary components of the sex pheromone of Matsucoccus feytaudi.

Oligomeric compounds formed from 2,5-xylidine (2,5-dimethylaniline) are potent enhancers of laccase production in Trametes versicolor ATCC 32745

Numerous chemicals, including the xenobiotic 2,5-xylidine, are known to induce laccase production in fungi. The present study was conducted to determine whether the metabolites formed from 2,5-xylidine by fungi could enhance laccase activity. We used purified laccases to transform the chemical and then we separated the metabolites, identified their chemical structure and assayed their effect on enzyme activity in liquid cultures of Trametes. versicolor. We identified 13 oligomers formed from 2,5-xylidine. (4E)-4-(2,5-dimethylphenylimino)-2,5-dimethylcyclohexa-2,5-dienone at 1.25×10−5 M was an efficient inducer, resulting in a nine-fold increase of laccase activity after 3 days of culture. Easily synthesized in one step (67% yield), this compound could be used in fungal bioreactors to obtain a great amount of laccases for biochemical or biotechnological purposes, with a low amount of inducer.

Hydroperoxides with zero, one, two or more carbonyl groups formed during the oxidation of N-dodecane

Abstract Experimental investigations and kinetic interpretation indicate the presence of several peroxidic species during the oxidation of n-dodecane. The oxidation was performed either in the gaseous phase in a flow system, or with a liquid/gas interaction in a bulb, because in many combustion processes liquid and gas coexist during preignition. The different peroxides were identified by mass spectrometry. Simple hydroperoxides with two oxygen atoms per molecule were observed at relatively low temperatures above 370 K with liquid and gas present. Ketohydroperoxides with three oxygen atoms per molecules are formed at ∼500 K. Di-, and even tri-, ketohydroperoxides with four or five oxygen atoms per molecule are also present under these experimental conditions at 518 K. Their formation is explained by isomerization reactions of alkoxy radicals OR −2H O bearing a carbonyl group; these stem from the decomposition of carbonyl-hydroperoxides. Depending on the experimental conditions, the nature of the hydroperoxides is different and the respective reactions of these species should be introduced in low temperature oxidation mechanisms. These reaction schemes should include the isomerization of peroxy radicals RO 2 and of ketoalkoxy radicals OR −2H O, leading to the formation of peroxides with several carbonyl groups.

Identification of the female sex pheromone of the Israeli pine bast scale Matsucoccus josephi

Abstract (2E,4E,8E)-4,6-dimethyl-2,4,8-decatrien-7-one ( I ) and (2E,4Z,8E)-4,6-dimethyl-2,4,8-decatrien-7-one ( II ), in an approximate ratio of 75:25, have been identified as the female sex pheromone components of Matsucoccus josephi. The structure of I and II was determined by mass spectrometric and infrared techniques and microreactions.

Multiresidue analysis of atrazine, diuron and their degradation products in sewage sludge by liquid chromatography tandem mass spectrometry

A multiresidue method has been developed to analyze atrazine (ATZ), diuron (DIU), and their major degradation products, desethylatrazine (DEA), desisopropylatrazine (DIA), and dichlorophenylmethylurea in sewage sludge. Liquid chromatography coupled to electrospray tandem mass spectrometry (LC–ESI-MS–MS) allowed, in the multiple-reaction monitoring mode, the simultaneous analysis of these pesticides in only one run after their extraction with ethyl acetate–dichloromethane 90:10 (v/v) and a cleanup on a Florisil column. Stable isotopically labeled ATZ and DIU were used as internal standards to overcome matrix effects during the pesticide quantification. Using fortified samples, the method gave rise to 86–115% as mean recovery values depending on the analyte. Limits of detection (LODs) and of quantification (LOQs) ranging from 0.3 (DIA) to 1.5 (DEA) μg kg−1 dw and from 0.4 (DIA) to 2.0 (DEA) μg kg−1 dw, respectively, were sufficient to achieve the monitoring of these molecules in sludge from wastewater treatment plants of the Ile-de-France region.

Isomeric differentiation of conjugated diene epoxides by polar [4 + 2+] Diels–Alder cycloaddition of acylium ions in an ion trap mass spectrometer

Abstract Polar [4 + 2 + ] Diels–Alder cycloaddition is performed using acylium cations as reagent ions for the characterization and differentiation of isomeric conjugated diene epoxides. The [M + RCO] + cycloadduct that is produced is unstable and dissociates not by retrocycloaddition, but by pathways favored by the epoxide ring that lead mainly to substituted pyrilium ions. These product ions display m/z ratios that correlate with either the epoxide (R 1 ) or diene (R 2 ) substituents, thus serving as structurally diagnostic ions for isomeric differentiation. The multiple tandem mass spectrometric (MS n ) capabilities of the quadrupole ion trap mass spectrometer (QITMS) were particularly useful for designing multistep procedures sometimes required for reagent-ion preparation and isolation. They were also appropriate when an additional collisionally induced dissociation (CID) stage was desired to provide further information on any product ion or for additional selectivity.