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

2. BETALAINS

3. BETALAMIC ACID

4. FORMATION OF THE BETALAMIC ACID

5. GENERAL STRUCTURES

6. EXTRACTION METHODS OF BATALAMIC
ACID
 Vegetable sources in aqueous solutions or purified water.
 Alkaline hydrolysis- Cyclo-DOPA-glucoside degradation, reverse reaction.

7. PURIFICATION OF BETALAMIC ACID
• Amine moiety of betalain structure
• Anionic exchange chromatography method(AEC)
• Trimethyl ammonium matrix
• 86% yield obtained

8. • 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)

9. 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 )

10. 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

11. 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-Trolox
White dot-Betalamic Acid

12. 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 control
Black dot-trolox
White dot- betalamic acid

13. NECLEOPHILIC CAPACITY OF THE
DEPRONATED FORM OF THE BETALAMIC
ACID
• 4 methyl catechol 4methylcatecholquinone new product at 530 nm

14. 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

15. ANTIOXIDANT ACTIVITY
• Reduction of Fe(III) INTO Fe(II) by batalamic acid at 593 nm
• Slopes=number of electrons reduced=2

16. 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.