Transcript Italian Red Passion and Annurca

6th International Conference on Polyphenols Applications
June 7 & 8, 2012 – Paris, France
POLYPHENOLS CONTENT IN APPLE ITALIAN
RED PASSION AND ANNURCA
MIHAELA BERINDE1,2, MAURIZIO RUZZI2, ROSARIO MULEO2, MARIA TOFANA1*, SONIA SOCACI1
1-Universitaty of Agricultural Science and Veterinary Medicine, Cluj-Napoca
2 - University of Tuscia, Viterbo, Italy
* Corresponding author: [email protected]
Purpose: To determine the polyphenols composition of apple (fruit pulp and leaf) from two varieties: Italian Red Passion and Annurca;
Preliminary evaluation of the antimicrobial activity of this apple varieties
Abstract
• Apples of seedlings, M3, M4 and M5 from the
experimental field of the University of Tuscia Italy,
belonging to the Italian Red Passion group are
investigated about the polyphenols content. These fruits
are characterized by red coloration of the pulp. In this
work we analyzed comparatively the polyphenols
content of apples M3 and M4 and leaves of apples M4
and M5. As control are used Annurca apples and
Annurca leaves.
•The total polyphenols content from all this samples were extracted with 3 types of solvents:
Methanol/Water/Acetic Acid:70:29:1, Acetone/Water/Acetic Acid:70:29:1 and
Methanol/
Acetone/Water /Acetic Acid: 60:20:19:1. The obtained extract was evaporated, lyophilized and
resuspended in methanol and then analyzed by HPLC. The HPLC analysis was performed,
simultaneously, at 5 wavelengths (280, 307, 316, 365 and 520 nm).
•Comparative analysis of HPLC data confirmed that Italian Red Passion extracts from both fruit
and leaf samples contained a larger array of compounds compared to Annurca samples. This
compounds, belong to different chemical groups, such as anthocyanin, hydroxycinnamic acids,
flavonols, procyanidins and dihydrochalcones. The better solvent systems for the chemical
analysis of fruit pulp and leaf samples are acetone and methanol respectively.
Introduction
• The assortment of polyphenols present in apple fruit is wide, but differs between flesh
and peel both in its range and in the total content (Wolfe et al., 2003). Typical apple fruit
phenolic acids are hydroxy-cinnamic acids, dihydro-chalcones, flavonols, anthocyanin,
flavanols and procyanidin (Tsao et al., 2003). The content changes of the main phenols
in apples and their contributions to the total antioxidant activity are not very clear.
•Numerous epidemiological studies, published toward the end of the twentieth century,
have provided evidence that increased consumption of fruit and vegetables provides
protection against cardiovascular diseases, various kinds of cancer, and other chronic
diseases. The risk of a heart attack has been shown to decrease by 49% for apple
consumers in diet at a daily amount of >110 g compared to those at <18 g.
•In fruits such as apple, red coloration of the skin is a much valued trait. In the last 20
years the interest on the polyphenolic content of the fruit is continuously growing up,
due to their health-related characteristics (Boyer and Liu, 2004); from this point of view,
red fleshed apple could be a varietal innovation with favorable health-related
characteristics and a strong appeal to consumers.
Functional compounds in apple:
organic acids, polyphenols, flavonoids
Materials and Methods
Results
HPLC chromatogram of anthocyanins (A520)
•The accessions were obtained from cross pollinated by a plant of red apple from the wild
germplasm Lazio in the experimental field at the farm of the University of Tuscia.
•In this work we used only apples M3 and M4 and leaves of apples M4 and M5. As control was
used Annurca apples and Annurca leaves.
Extract preparation:
Lyophilized material
Solvent extraction
 Methanol/Water/Acetic acid (70:29:1)
Acetone/Water/Acetic acid (70:29:1)
Methanol/Acetone/Water/Acetic acid (60:20:19:1)
Compounds identified
Graphical representation of HPLC results
from methanol extracts of M4 fruit pulp
Dried samples were resuspended in pure MetOH
HPLC Analysis was performed using a HPLC-DAD apparatus at 5 wavelengths (280, 307, 316, 365 and 520
nm), following the method used by Jeffrey et al. (1999)
Bacterial antimicrobial
susceptibility plate test:
modified Kirby-Bauer Method
ANTIMICROBIAL ACTIVITY OF EXTRACTS
FROM APPLE ITALIAN RED PASSION AND ANNURCA
Bacterial antimicrobial susceptibility
liquid test: Broth Dilution Method
Growth inhibition obtained using acetone
extracts from fruit pulp of different Italian Red
Passion genotypes (M3, M4) and Annurca
Higher antimicrobial activity was obtained
using fruit pulp extracts of M4 genotype,
independently from the solvent system
GROWTH INHIBITION
INHIBITION ZONE DIAMETER
Literature
Wolfe K, Liu RH. (2003), “ Apple peels as a value-added food ingredient”. J Agric Food Chem. 2003, 51:1676–1683.
Tsao Rong, Yang Raymond, Young J. Christopher and Zhu Honghui, (2003), ”Polyphenolic Profiles in Eight Apple Cultivars Using High
Performance Liquid Chromatography (HPLC)”, J. Agric. Food Chem., 51: 6347-6353.
Alberto María Rosa, Matías Andrés, CanavosioRinsdahl, Manca de NadraMaría Cristina,(2006), “Antimicrobial effect of polyphenols from apple
skins on human bacterial pathogens”, Electronic Journal of Biotechnology ISSN: 0717-3458 Vol.9 No.3, Special Issue, by Pontificia
Universidad Católica de Valparaíso – Chile.
Boyer J.; Liu, R.H,(2004), “Apple phytochemicals and their health benefits”, Nutrition Journal, 3: 5.
Kondo Satoru, Tsuda Kazuhiko, Muto Norio, Ueda Jin-etsu, (2002) “Antioxidative activity of apples skin or flesh extracts associated whit fruit
development on selected apple cultivars”,ScientiaHorticulturae, 96: 177-185.
Mari Angela, Tedesco Idolo, Nappo Annunziata, Russo Gian Luigi, Malorni Antonio, Carbone Virginia, (2010) “Phenolic compound
characterization and anti-proliferative activity of ‘‘Annurca” apple, a southern Italian cultivar”, Food Chemistry, 123: 157–164.
D’AngeloStefania, Cimmino Amelia, Raimo Marianna, Salvatore Anna, Zappia Vincenzo , and GallettiPatrizia, (2007) “Effect of Reddening–
Ripening on the Antioxidant Activity of Polyphenol Extracts from Cv. ‘Annurca’ Apple Fruits”,J. Agric. Food Chem, 55:9977–9985.
SOLVENT
E. coli DH 5α
B.subtilis PB19
P.aeruginosa
SOLVENT
(cm)
SAMPLE
SAMPLE
(%)
25 µl
50 µl
M3
none
25.20±0.00172
M4
19.52±0.0053
67.87±0.0004
0.450±0.0
Annurca
none
68.40
0.275±0.00125
0.250±0.0
M3
none
25.02±0.01533
M4
0.550±0.005
0.525±0.00125
M4
35.00±0.0156
53.32±0.0013
Annurca
0.375±0.0
0.350±0.00125
Annurca
25.65±0.01346
59.18±0.00019
M3
0.225±0.00125
0.225±0.00125
M4
0.400±0.0
0.400±0.0
M3
none
21.66±0.00274
Annurca
0.400±0.0
0.400±0.0
M4
35.55±0.0008
60.25±0.0016
0.0
0.0
Annurca
33.90±0.00273
61.84±0.00047
24h
48h
M3
0.325±0.00125
0.300±0.0
M4
0.600±0.0
0.575±0.00125
Annurca
0.450±0.005
M3
Control(methanol)
Acetone
Methanol
Mix