Structural analysis and profiling of phenolic secondary metabolites of Mexican lupine species using LC-MS techniques.

Flavonoid glycoconjugates from roots and leaves of eight North America lupine species (Lupinus elegans, Lupinus exaltatus, Lupinus hintonii, Lupinus mexicanus, Lupinus montanus, Lupinus rotundiflorus, Lupinus stipulatus, Lupinus sp.), three Mediterranean species (Lupinus albus, Lupinus angustifolius, Lupinus luteus) and one species from South America domesticated in Europe (Lupinus mutabilis) were analyzed using two LC/MS systems: low-resolution ion trap instrument and high-resolution quadrupole-time-of-flight spectrometer. As a result of the LC/MS profiling using the CID/MS(n) experiments structures of 175 flavonoid glycoconjugates found in 12 lupine species were identified at three confidence levels according to the Metabolomic Standard Initiative, mainly at level 2 and 3, some of them were classified to the level 1. Among the flavonoid derivatives recognized in the plant extracts were isomeric or isobaric compounds, differing in the degree of hydroxylation of the aglycones and the presence of glycosidic, acyl or alkyl groups in the molecules. The elemental composition of the glycoconjugate molecules was established from the exact m/z values of the protonated/deprotonated molecules ([M+H](+)/[M-H](-)) measured with the accuracy better than 5 ppm. Information concerning structures of the aglycones, the type of sugar moieties (hexose, deoxyhexose or pentose) and, in some cases, their placement on the aglycones as well as the acyl substituents of the flavonoid glycoconjugates was achieved in experiments, in which collision-induced dissociation was applied. Flavonoid aglycones present in the studied O-glycoconjugates were unambiguously identified after the comparison of the pseudo-MS(3) spectra with the spectra registered for the standards. Isomers of flavonoid glycoconjugates, in which one or two sugar moieties were attached to 4'- or 7-hydroxyl groups or directly to the C-6 or C-8 of the aglycones, could be distinguished on the basis of the MS(2) spectra. However, the collision energy applied in the CID experiments had to be optimized for each group of the compounds and there were no universal settings that allowed the acquisition of structural information for all the compounds present in the sample. Information obtained from the flavonoid conjugate profiling was used for the chemotaxonomic comparison of the studied lupine species. A clear-cut discrimination of the Mediterranean and North American lupines was obtained as a result of this analysis.

LC-MSMS profiling of flavonoid conjugates in wild Mexican lupine, Lupinus reflexus.

Profiles of flavonoid conjugates present in the root and leaf tissues of the Mexican wild lupine, Lupinus reflexus, were established using two LC-MSMS systems in the positive and negative ion modes. The ion trap mass spectrometer and quadrupole time-of flight instrument provided sequential MS(n) spectra and MSMS spectra with accurate m/z values of [M + H](+) and [M - H] (-) ions, respectively. Sixty-two flavone and isoflavone glycoconjugates were found and tentatively identified. Numerous isomeric or isobaric compounds with the same molecular mass could be differentiated. Isomeric di- and mono glucosides of biochanin A, genistein, 2'-hydroxygenistein, luteone, and 2,3-didehydrokievitone were distinguished on the basis of relative abundances of product ions. The studied flavonoid glycoconjugates were acylated with dicarboxylic aliphatic acids and their methyl esters at either the aglycone or glycosidic moiety.

Composición y concentración de alcaloides en Lupinus Exaltatus zucc. durante su crecimiento y desarrollo / Alkaloid composition and concentration in Lupinus Exaltatus zucc. during its growth and development / Composição e concentração de alcaloides em Lupinus Exaltatus zucc. durante seu crescimento e desenvolvimento

Por sus características nutricionales Lupinus exaltatus es considerado una fuente alternativa de alimento para animales en zonas templadas, pero la presencia de alcaloides quinolizidínicos limita su consumo. Se determinó la composición y concentración de alcaloides en hojas, tallos, flores y frutos inmaduros de L. exaltatus durante cinco etapas fenológicas para proponer alternativas de uso y manejo de esta especie como forraje. Se estableció un cultivo en macetas y en cada etapa fenológica se tomaron seis plantas que se separaron en sus diferentes órganos, para ser deshidratados y analizar su composición y contenido de alcaloides por cromatografía de gases capilar-espectrometría de masas (CG-EM). En cada etapa y órganos se identificaron los alcaloides quinolizidínicos lupanina, 3-b-hidroxilupanina, afilina, epiafilina, dehidro-oxoesparteína y a-isolupanina. Aunque el perfil permaneció constante, la concentración de alcaloides totales en los diferentes órganos fluctuó significativamente (0,31-2,1%) en función de las etapas de crecimiento. Tras cinco etapas de crecimiento y desarrollo el menor contenido promedio de alcaloides se encontró en tallos (0,63%) mientras que en frutos inmaduros (vainas verdes) se encontró el más alto (1,95%). La lupanina resultó ser el compuesto individual mayoritario en tallos y frutos. Aunque no se detectaron alcaloides potencialmente tóxicos y teratogénicos (esparteína citisina, anagirina y amodendrina), los resultados indican que el mayor riesgo de intoxicación del ganado por consumo de L. exaltatus podría ocurrir en las etapas de formación de vainas y fructificación, debido a la alta concentración de alcaloides totales y mayor abundancia de lupanina en frutos inmaduros.

Due to its nutritional characteristics, Lupinus exaltatus is considered as an alternative source of animal feed in temperate regions. However, the presence of quinolizidine alkaloids in different plant organs limits its consumption. The composition and concentration of alkaloids in leaves, stems, flowers, and inmature fruits of L. exaltatus was determined in five different phenological stages with the final purpose of suggesting alternatives for its management and use as forage. Plants were sowed in pots and at each phenological stage six plants were harvested and separated in roots, stems, leaves, flowers, and inmature pods that were air dried to constant weight. Each organ was analyzed for alkaloid composition and content by capillary gas chromatography- mass spectrometry (GC-MS). In each phenological stage and in all organs the alkaloid profile was characterized by the presence of six major alkaloids: epiaphylline aphylline, a-isolupanine, lupanine, dehydro-oxosparteine and 3-b-hydroxylupanine. Although the alkaloid profile was constant, the total concentration in the different organs showed a significant variation (0.31-2.1%) in the different phenological stages. In general, after the five growth stages the stems showed the lowest average total alkaloid concentration (0.63%) whereas inmature pods had the highest total alkaloids (1.95%). Lupanine was the major alkaloid in stems and fruits. Although alkaloids with the highest toxicity (sparteine, anagyrine and ammodendrine) were not detected, the results indicate that the major risk for intoxication could occur during pod growth and seed ripening, due to high total alkaloid content and larger abundance of lupanine in immature fruits.

Por suas características nutricionais Lupinus exaltatus é considerado uma fonte alternativa de alimento para animais em regiões temperadas, mas a presença de alcalóides quinolizidínicos limita seu consumo. Determinou-se a composição e concentração de alcaloides em folhas, caules, flores e frutos imaturos de L. exaltatus durante cinco etapas fenológicas para propor alternativas de uso e manejo desta espécie como forragem. Estabeleceu-se um cultivo em vasos e em cada etapa fenológica foram colhidas seis plantas e separardas em seus diferentes órgãos, para serem deshidratados e para analizar sua composição e conteúdo de alcalóides por cromatografia de gases capilar-espectrometria de massas (CG-EM). Em cada etapa e órgãos foram identificados os alcalóides quinolizidínicos lupanina, 3-b-hidroxi lupanina, afilina, epiafilina, dehidro-oxoesparteína e a-isolupanina. Ainda que o perfil tenha permanecido constante, a concentração de alcaloides totais nos diferentes órgãos flutuou significativamente (0,31-2,1%) em função das etapas de crescimento. Depois de cinco etapas de crescimento e desenvolvimento o menor conteúdo médio de alcaloides foi encontrado em caules (0,63%) enquanto que em frutos imaturos (vagens verdes) se encontrou o mais alto (1,95%). A lupanina resultou ser o composto individual maioritário em caules e frutos. Ainda que não se detectaram alcaloides potencialmente tóxicos e teratogénicos (esparteína citisina, anagirina e amodendrina), os resultados indicam que o mayir risco de intoxicação do gado por consumo de L. exaltatus poderia ocorrer nas etapas de formação de vagens e frutificação, devido a alta concentração de alcaloides totais e maior abundância de lupanina em frutos imaturos.

Evaluación de lodos residuales como abono orgánico en suelos volcánicos de uso agrícola y forestal en Jalisco-México / Evaluation of sewage sludge as an organic fertilizer in volcanic soils of agricultural and forestal use in Jalisco-Mexico

La posibilidad de reutilizar materiales orgánicos ricos en nutrientes hace de la aplicación de lodos residuales en suelos agrícolas y forestales una alternativa importante. Se evaluaron diferentes dosis de lodos de aguas residuales sanitarias como abono orgánico en la productividad de maíz y sobre el desarrollo inicial de Pinus douglasiana. Los experimentos se establecieron en un suelo volcánico ubicado en la zona centro de Jalisco, México. En el cultivo de maíz se evaluaron 10 y 20 t-ha-¹ de lodos deshidratados, así como 10 y 20 t-ha-¹ de compost de lodos mezclados con residuos de jardinería. Los materiales se distribuyeron homogéneamente sobre el suelo bajo un diseño experimental de bloques al azar con cinco repeticiones. Se evaluó el rendimiento de grano y de forraje. El rendimiento de grano en los tratamientos con lodos residuales y compost se incrementó significativamente, en 18 y 22 por ciento respectivamente, con respecto al control. En la plantación de Pinus douglasiana se evaluaron 00,30, 60 y 100 g de lodos por árbol. A los 14 meses de la plantación se registró una supervivencia de hasta 83 por ciento en las parcelas con mayor dosis de lodos mientras que en el control la supervivencia fue de 67 por ciento. La altura y diámetro del tallo se incrementaron en ~18 por ciento con la mayor dosis de lodos, en comparación con el control. La aplicación de lodos de aguas residuales sanitarias como abono orgánico mejoró la producción de maíz y el crecimiento inicial de Pinus douglasiana en suelos volcánicos.