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Chemical and biological assays for quantification of major plant secondary metabolites

Published online by Cambridge University Press:  27 February 2018

Harinder P.S. Makkar
Harinder P.S. Makkar*
Affiliation:
Animal Production and Health Section, Joint FAO/IAEA Division, International Atomic Energy Agency, Vienna, Austria
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Summary

During grazing, animals interact with many compounds in the diet. Plant secondary metabolites (PSM) such as tannins, saponins, cyanogenic glycosides, mimosine. could have wide ranging effects –beneficial or harmful, depending on the nature, quantity in the plant, and amount and rate of ingestion of the PSM. A wide range of methods is available to quantify these PSM. The methods presented for tannins are based on the oxidation-reduction, metal complexing, polyethylene glycol binding and protein precipitation/binding principles of tannins. A tannin bioassay based on in vitro gas production using a medium containing rumen microbes, and incubation of the plant sample with and without polyethylene glycol (a tannin-inactivating agent) is also discussed. Saponins have haemolitic activity, and a qualitative and a quantitative assay are based on this property are described, in addition to a spectrophotometric assay. Cyanogenic glycosides are measured using picric acid methods with and without distillation. The methods described for mimosine determination are based on its reaction with ferric chloride and diazotised p-nitroaniline. Alkaloids are a chemically heterogeneous group of compounds. Therefore, it is difficult to assay alkaloids using a single method. An approach, for qualitative assessment of alkaloids in feeds and forages, based on the use of various spray reagents on the developed TLC plate or a paper chromatographic paper is presented.

Resumen

Resumen

Durante el pastoreo, los animales interactúan con muchos compuestos en la dieta. Los metabolitos secundarios de las plantas (MSP) tales como los taninos, saponinas, glicosidos cyanogénicos y mimosina pueden tener un amplio rango de efectos –benéficos ó dañinos dependiendo de la naturaleza y cantidad en la planta así como la cantidad y tasa de ingestión de los MSP. Una amplia variedad de métodos están disponibles para cuantificar estos MSP. Los métodos presentados para taninos están basados en la oxidación-reducción, complejamiento de metales, acoplamiento de polietilen glicol y principios de precipitación/acoplamiento de taninos. También se discute un bioensayo de taninos basado en la producción de gas in vitro que utiliza un medio que contiene microbios ruminales e incubación de muestras de plantas con o sin polietilen glicol (un agente inactivador de taninos). Las saponinas tienen actividad hemolítica y se describen un ensayo cualitativo y cuantitativo basado en esta propiedad, además de un ensayo espectrofotométrico. Los glicósidos cianogénicos son medidos usando métodos de ácido pícrico con y sin destilación. Los métodos descritos para la determinación de mimosina están basados en su reacción con el cloruro férrico y pnitroanilina diazotisada. Los alcaloides son un grupo de compuestos químicamente heterogeneos. Por lo tanto, es difícil realizar pruebas de alcaloides usando un solo método. Se presenta una forma de enfrentar esto, para la determinación cualitativa de alcaloides en alimentos y forrajes que se basa en el uso de varios reactivos atomizados en el plato TLC desarrollado ó en un papel cromatográfico.

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BSAP Occasional Publication , Volume 34: Herbivores: Assessment of intake, digestibility and the roles of secondary compounds , 2006 , pp. 235 - 249
Copyright
Copyright © British Society of Animal Science 2006

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