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The total free amino acid content was determined by the ninhydrin test,
according to the method of ircelj et.al. (2005). A known weight of dry
powdered plant material of each sample was extracted in 80% ethanol and
centrifuged at 8,000g for 15min. The supernatant containing the alcohol-soluble
fraction was collected. A reaction mixture, consisting of 0.1ml extract, 0.9 ml of
double-distilled water and 1ml of ninhydrin reagent was shaken vigorously and
then heated in a boiling water bath for about 20 min and added to 5ml diluents
(equal volumes of water and n-propanol). After cooling the tubes to room
temperature, the color intensity was measured at 570nm. The blank set was
prepared by adding 0.1ml of 80% ethanol in place of sample extract. The
standard curve was prepared by using leucine as a standard amino acid. The
results were expressed in terms of mg of amino acid per g of dry tissue.
required for catalyzing the conversion of 1mmol H2O2 into water per min (U =
1mM of H2O2 reduction min1 mg1 of protein) according to the method of Aebi
(1984).
ajout de l
acide sulfurique (aprs destruction des sucres complexes).La concentration des sucres totaux
dans les chantillons doser a t calcule par rapport la calibration au glucose. Une
calibration l
Dosage des sucres totauxLe dosage des sucres totaux solubles est ralis par la mthode de
Dubois et al.(1956).II.3.6.1 PrincipeLes oses sont stables en milieu acide. Cependant s
ils sont chauffs en milieu acide concentr, ils donnent des furfuraldhydes par cyclisation et
dshydratation. Les furfurals etses drivs ont la proprit de se condenser avec le phnol
pour former des complexesmarron.II.3.6.2 Mode opratoireDans une srie de tubes essai, 25
l d
acide sulfurique concentre (H2SO4). Le mlange est chauff aubain marie 100 C pendant
5 mn. Aprs refroidissement dans la glace fondante, la densitoptique est mesure 490 nm
contre un blanc dans lequel 25 l d
alcool 80 % remplacel
extrait brut. Un talon est construit grce une gamme (0 35 mg/ml) de concentrationd
une solution mre de glucose 1 mg/ml (Fig. 8). Les quantits sont exprimes en mg/g de poids
frais et dtermines par la formule suivante