A sensitive and selective spectrophotometric method for the estimation of the toxic factors mimosine and 3-hydroxy-4-(1H)-pyridone (DHP) has been developed based on the intense yellow-colored azodye formed with p-nitroaniline which showed a sharp absorption maximum at 400 nm. The method was optimized based on relative sensitivity of the reaction with various aromatic primary amino compounds and under different conditions of pH. Interference from a variety of structurally related compounds and phenols was tested and found insensitive to this method. The molar extinction coefficient at 400 nm for the azodye formed with mimosine was 5.31 × 104 M-1 cm-1 and that for DHP was 1.699 × 104 M-1 cm-1. The applicability of the method was tested using different plant extracts and recovery was found to be at 100 ± 0.3%. The method is suitable for accurate estimation of both mimosine and DHP after paper chromatographic separation of extracts of different biological samples. © 1993 Academic Press. All rights reserved.

Kalyanaraman Lalitha

3 HYDROXY 4 PYRIDONE

MIMOSINE

unclassified drug

article

nonhuman

plant leaf

priority journal

spectrophotometry

LEUCAENA LEUCOCEPHALA

LEUCOCEPHALA

IIT Madras is a public technical and research university located in Chennai, Tamil Nadu. Founded in 1959, it is recognised as an Institute of National Importance.

IIT Madras has been ranked as the top engineering institute in India for four years in a row by the National Institutional Ranking Framework of the MHRD

It currently offers undergraduate, postgraduate and research degrees across 16 disciplines in Engineering, Sciences, Humanities and Management. About 596 faculty belonging to science and engineering departments and centres of the Institute are engaged in teaching, research and industrial consultancy.

IIT Madras

Analytical Biochemistry

Our observations on the growth stimulatory nature of mimosine, (β-(3-hydroxy-4-pyridon-1-yl)-L-alanine), the toxic non-protein plant amino acid, in some model experimental systems, warranted sensitive and selective routine estimations. For the determination of both mimosine and DHP, an indirect spectrophotometric method was developed based on their individual reaction with known excess of DZSAM and by estimating the remaining DZSAM with N-(1-naphthyl)ethylene-diamine (NEDA). The resultant decrease in the secondary coupled product was measured at 540 nm. On equimolar basis, DHP had 40% of the reactivity of mimosine while interference from other relevant compounds was 15-35%. The determination of mimosine and DHP in tissue samples under different physiological conditions was effected after paper chromatographic separation of mimosine and DHP with distinctly differing Rf, from other compounds. The indirect method is superior in terms of absolute selectivity, sensitivity and ease of applicability with linear decreases in absorbance, proportional to increasing concentrations of mimosine from 0.1 to 0.75 μM or DHP from 0.2 to 1.5 μM and with recoveries of 99.2 to 100.5%. © 2006 Springer-Verlag.

Amino Acids

Selective determination of mimosine and its dihydroxypyridinyl derivative in plant systems

A simple and sensitive indirect spectrophotometric method is described for the determination of mimosine based on its reaction with diazotized sulfanilamide (DZSAM). DZSAM couples with N-(1-naphthyl)ethylenediamine (NEDA) forming a pink colored azodye, absorbing maximally at 540 nm (εmax=27 mM-1 cm-1). In the present method, mimosine was first reacted with known excess of DZSAM and the unreacted DZSAM was determined by coupling with NEDA. The reaction of mimosine with DZSAM proceeded optimally at neutral pH. The decrease in absorbance of the DZSAM-NEDA-coupled product obeyed Beer's law in the concentration range of 0.005-0.15 μg ml-1 of mimosine. The present method was applied to estimate mimosine in plant extracts containing lesser than 0.05 μg ml -1 with recovery at 99±0.41%. The method described is superior to other reported methods in terms of ease of adaptability and sensitivity. © 2003 Elsevier B.V. All rights reserved.

Talanta

Determination of mimosine by a sensitive indirect spectrophotometric method

Glycohydrolases assume significance in the metabolism of biological systems and have important industrial applications in the areas of pharmaceuticals, food, and medicine. Glycosidases were screened in germinating seeds, and attempts were made to enhance their levels. Screening of glycosidases in the seedlings during a 72 h germination period revealed higher levels of β-glucosidase and β-galactosidase in Trigonella foenum- graecum compared to Cicer arietinum and Vigna radiata. Activity of β- galactosidase was in general higher than that of β-glucosidase in all the seedlings tested. During growth, exposure of the seedlings to an allelochemical, mimosine, at 0.1 mM resulted in the enhancement of enzyme levels by 50% in the seedlings of T. foenum-graecum, whereas the addition of mimosine to the assay medium in vitro did not affect the enzyme activities. Hydrolytic activity was enhanced by addition of glycerol in the medium up to 0.1 M in the case of β-glucosidase and with 0.05 M in the case of β- galactosidase. In general, the hydrolytic rate was higher by about 30% in the seedlings exposed to mimosine compared to that of the control. Concomitant enhancement in the rates of transgalactosidation by 51% and transglucosidation by 23% was also noted, underscoring the relevance of plant glycohydrolases for appropriate applications.

Journal of Agricultural and Food Chemistry

Enhancement of β-glucosidase and β-galactosidase of Trigonella foenum- graecum by exposure to the allelochemical mimosine

Spectrophotometric Determination of Mimosine and 3-Hydroxy-4-(1H)-pyridone - The Toxic Principles of Leucaena leucocephala

Journal	Analytical Biochemistry
ISSN	00032697
Open Access	No