Fernando Gandía-Herrero,* Josefa Escribano, and Francisco García-Carmona

PURIFICATION AND ANTIRADICAL PROPERTIES OF THE STRUCTURAL UNIT OF

BETALAINS

By - Summervir Cheema

BETALAINS

BETALAMIC ACID

FORMATION OF THE BETALAMIC ACID

GENERAL STRUCTURES

EXTRACTION METHODS OF BATALAMIC ACID

Vegetable sources in aqueous solutions or purified water. Alkaline hydrolysis- Cyclo-DOPA-glucoside degradation, reverse reaction.

PURIFICATION OF BETALAMIC ACID• Amine moiety of betalain structure

• Anionic exchange chromatography method(AEC)

• Trimethyl ammonium matrix

• 86% yield obtained

• 10 ml of the 250µM betanin hydrolysis medium

• 100µL of the 250µM betanin hydrolysis medium

PURIFICATION OF BETALAMIC ACID

(A) 1 ml column (B) 5 ml column

Wavelength = 424nm(dark line), 280nm(dotted line)

HPLC ANALYSIS OF BATALAMIC ACID INJECTION VOL- 20 Microliter of 80 micromolar betalamic acid

RETENTION TIME= 14.59 MIN

MAX WAVELENGTH ABSORBANCE= 407 nm with MeCN ( acetonitrile )

UV-VIS SPECTRUM OF BETALAMIC ACID IN WATER

25 µm of Betalamic Acid Soln. in H2O.

WAVELENGTH MAX= 424 nm

Molar absorptivity /epsilon value=27000 M^-1. cm^-1

FREE RADICAL SCAVENGING ACTIVITY OF BETALAMIC ACID

Colored radical-ABTS[2,2’-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid)

Anti radical-Trolox ( 6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid)

ABTS decolorization assay

Dec in abs at 734 nm

TEAC(Trolox equivilent antiradical capacity) value= 2.7+/- 0.2

Black dot-TroloxWhite dot-Betalamic Acid

PH EFFECT ON THE FREE RADICAL SCAVENGING ACTIVITY OF BETALAMIC ACID

buffers sodium acetate ph (3.5-5.5), sodium phosphate ph (5.5-8.5)

Protonation equilibrium

Square dot-water controlBlack dot-troloxWhite dot- betalamic acid

NECLEOPHILIC CAPACITY OF THE DEPRONATED FORM OF THE

BETALAMIC ACID• 4 methyl catechol 4methylcatecholquinone new product at 530 nm

PH EFFECT ON THE NEUCLEOPHILIC CAPACITY OF BATALAMIC ACID

• In the reaction medium-100 µM betalmic acid+200 µM 4-methylcatechol+Tyrosinase+50mM buffer-NO QUINONE

• Kapp ( apparent rate constant ) = high=higher pH,low=lower Ph

• pKa=6.8 for deprotonated form of betalmic acid

ANTIOXIDANT ACTIVITY• Reduction of Fe(III) INTO Fe(II) by batalamic acid at 593 nm

• Slopes=number of electrons reduced=2

APPLICATIONS Betalains free forms can be used in pharmaceutical industry in order to obtain the new

betalains.

In humans, betalains protect the red blood cells from oxidative damage and hemolysis.

Very low concentrations of betalains inhibit skin and liver tumor formation in mice.