Fernando Gandía-Herrero,* Josefa Escribano, and Francisco García-Carmona PURIFICATION AND ANTIRADICAL PROPERTIES OF THE STRUCTURAL UNIT OF BETALAINS By - Summervir Cheema
Fernando Gandía-Herrero,* Josefa Escribano, and Francisco García-Carmona
PURIFICATION AND ANTIRADICAL PROPERTIES OF THE STRUCTURAL UNIT OF
BETALAINS
By - Summervir Cheema
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