Analysis of glycosylated flavonoids extracted from sweet-cherry stems, as antibacterial agents against pathogenic Escherichia coli isolates

  • Alfredo Aires Centre for the Research and Technology for Agro-Environment and Biological Sciences, CITAB, Universidade de Trás-os-Montes e Alto Douro, UTAD, Quinta de Prados, 5000-801 Vila Real, Portugal.
  • Carla Dias Animal and Veterinary research Centre, CECAV, Universidade de Trás-os-Montes e Alto Douro, UTAD, Quinta de Prados, 5000-801 Vila Real, Portugal
  • Rosa Carvalho Agronomy Department, Universidade de Trás-os-Montes e Alto Douro, UTAD, Quinta de Prados, 5000-801 Vila Real, Portugal.
  • Maria José Saavedra Animal and Veterinary research Centre, CECAV, Universidade de Trás-os-Montes e Alto Douro, UTAD, Quinta de Prados, 5000-801 Vila Real, Portugal
Keywords: Flavonoids, agro-food wastes, enhanced extraction, antimicrobial agents, pathogenic bacteria


The aim of this study was to evaluate the bioactivity of flavonoids extracted from sweet-cherry stems, which is often used by traditional medicine to treat infections from gastro-intestinal and urinary but without any consistent scientific evidences, moreover the information about the class of phenolics, their content and the potential bioactivity of such material is very scarce. Thus, in this context, we set a research study in which we evaluate the profile and content of phenolics extracted from sweet-cherry stems through a conventional (70ºC and 20 minutes) and ultrasound assisted extraction (40 kHz, room temperature and 20 minutes) methods. After, the extracts were phytochemical characterized by HPLC-DAD-UV/VIS, and assayed trough the in vitro minimum inhibitory concentration (MIC) bioassay, against Escherichia coli isolates. Simultaneously the total antioxidant activity were measured using the assay of 2,2′-azinobis-3-ethylbenzothiazoline-6-sulphonate (ABTS●+) radical cation. Our results showed that sweet-cherry stems presented higher content of sakuranetin, ferulic acid, p-coumaric acid, p-coumaroylquinic acid, chlorogenic acid and it´s isomer neochlorogenic acid. Their average levels were highly affected by the extraction method (p<0.001) used. The same trend was observed for total antioxidant activity and MIC values. The extracts produced under ultrasound presented both higher total antioxidant activity and lower minimum inhibitory concentration. The statistical analyses of our results showed a significant correlation (p<0.01) of total antioxidant activity and minimum inhibitory concentration with phenolics in the extracts studied. Thus, we can conclude that cherry stems can be further exploited to purify compounds and produced coproducts with enhanced biological added valuefor pharmaceutical industry.


Ajil CM, Brar SK, Verma M, Tyagi RD, Godbout S, Valéro JR (2012) Bio-processing of agro-byproducts to animal feed. Crit Rev Biotechnol 32: 382-400. doi: 10.3109/07388551.2012.659172.

Alfieri M, Hidalgo A, Berardo N, Redaelli RJ (2014) Carotenoid composition and heterotic effect in selected Italian maize germplasm. Cereal Sci 59: 181-188. doi:10.1016/j.jcs.2013.12.010.

Alighourchi HR, Barzegar M, Sahari MA, Abbasi S (2013) Effect of sonication on anthocyanins, total phenolic content, and antioxidant capacity of pomegranate juices. Int Food Res J 20: 1703-1709.

Ashokkumar, M, Sunartio, D, Kentish, S, Mawson, R, Simons, L, Vilkhu, K, Versteeg, C K (2008) Modification of food ingredients by ultrasound to improve functionality: A preliminary study on a model system. Innov Food Sci Emerg Technol 9: 155-160. doi:10.1016/j.ifset.2007.05.005

Ballisteri G, Continella A, Gentile A, Amenta M, Fabroni S, Rapisarda P (2013) Fruit quality and bioactive compounds relevant to human health of sweet cherry (Prunus avium L.) cultivars grown in Italy. Food Chem 140: 630–638. doi:10.1016/j.foodchem.2012.11.024

Bansode RR, Randolph P, Hurley S, Ahmedna M (2012)

Evaluation of hypolipidemic effects of peanut skin-derived polyphenols in rats on Western-diet. Food Chem 135: 1659-1666. doi: 10.1016/j.foodchem.2012.06.034.

Coates ARM, Halls G, Hu Y (2011) Novel classes of antibiotics or more of the same? Brit J Pharmacol 163: 184-194. doi: 10.1111/j.1476-5381.2011.01250.x

Da Silva GJ, Mendonca N (2012) Association between antimicrobial resistance and virulence in Escherichia coli. Virulence 3: 18-28. doi: 10.4161/viru.3.1.18382

Denev P, Kratchanova M, Cizb M, Lojek A, Vasicek O, Nedelcheva P, Blazheva D, Toshkova R, Gardeva E, Yossifova L, Hyrsl P, Vojtek L (2014) Biological activities of selected polyphenol-rich fruits related to immunity and gastrointestinal health. Food Chem 157: 37-44. doi: 10.1016/j.foodchem.2014.02.022.

Fernando IDNS, Abeysinghe DC, Dharmadasa RM (2013) Determination of phenolic contents and antioxidant capacity of different parts of Withania somnifera (L.) Dunal from three different growth stages. Ind Crop Prod 50: 537-539. doi:10.1016/j.indcrop.2013.08.042

Jorgensen JH, Turnidge JD, Washington JA (2009) Antibacterial susceptibility tests: dilution and disk diffusion methods In: P R Murray, E J Baron, M A Pfaller, F C Tenover, and R H Yolken (Eds), Manual of clinical microbiology (7th ed), (pp 1526e1543).

Kuete V, Alibert-Franco S, Eyong KO, Ngameni B, Folefoc GN, Nguemeving JR, Tangmouo JG, Fotso GW, Komguem J, Ouahouo BM, Bolla, JM, Chevalier J, Ngadjui BT, Nkengfack AE, Pages JM (2011) Antibacterial activity of some natural products against bacteria expressing a multidrug-resistant phenotype. Int J Antimicrob Ag 37: 156–161. doi: 10.1016/j.ijantimicag.2010.10.020.

Lobo V, Patil A, Phatak A, Chandra N (2012) Free radicals, antioxidants and functional foods: Impact on human health. Pharmacogn Rev 4: 118–126. doi: 10.4103/0973-7847.70902.

Murray PR, Boron EJ, Pfaller MA, Tenover F, Yolken RH (1999) ASM Press: Washington, DC 876 pp.

Park HL, Yoo Y, Hahn TR, Bhoo SH, Lee SW, Cho MH (2014) Antimicrobial activity of UV-induced phenylamides from rice leaves. Molecules 19: 18139-18151. doi: 10.3390/molecules191118139.

Sánchez-Vioque R, Polissiou M, Astraka K, de los Mozos-Pascual M, Tarantilis P, Herraiz-Peñalver D, Santana-Méridas O (2013) Polyphenol composition and antioxidant and metal chelating activities of the solid residues from the essential oil industry. Ind Crop Prod 49: 150-159. doi:10.1016/j.indcrop.2013.04.053

Sargın SA, Akçicek E, Selvi S (2013) An ethnobotanical study of medicinal plants used by the local people of Alaşehir (Manisa) in Turkey. J Ethnopharmacol 150: 860-874. doi: 10.1016/j.jep.2013.09.040.

Sarker SD, Nahar L, Kumarasamy Y (2007) Microtitre plate-based antibacterial assay incorporating resazurin as an indicator of cell growth, and its application in the in vitro antibacterial screening of phytochemicals. Methods 42: 321-324. doi: 10.1016/j.ymeth.2007.01.006.

Schneider Y, Zahn S, Hofmann J, Wecks M, Rohm H (2006) Acoustic cavitation induced by ultrasonic cutting devices: A preliminary study. Ultrason Sonochem 13: 117-120. doi:10.1016/j.ultsonch.2005.09.002.

Schoevaerdts D, Bogaerts P, Grimmelprez A, de Saint-Hubert M, Delaere B, Jamart J, Swine C, Glupczynski Y (2011) Clinical profiles of patients colonized or infected with extended-spectrum beta-lactamase producing Enterobacteriaceae isolates: a 20 month retrospective study at a Belgian University Hospital. BMC Infect Dis 11: 12. doi: 10.1186/1471-2334-11-12.

Shimizu T, Lin F, Hasegawa M, Nojiri H, Yamane H, Okada K (2012) The potential bio-production of the pharmaceutical agent sakuranetin, a flavonoid phytoalexin in rice. Bioeng 3: 352-357. doi: 10.4161/bioe.21546.

Simões, M, Bennett RN, Rosa, EAS (2009) Understanding antimicrobial activities of phytochemicals against multidrug resistant bacteria and biofilms. Nat Prod Rep 26: 746-757. doi: 10.1039/b821648g.

Tiwari BK, Patras A, Brunton N, Cullen PJ, O’Donnell CP (2010) Ultrasound assisted extraction of bioactive compounds from Nephelium lappaceum L. fruit peel using central composite face centered response surface design. Ultra Sonochem 17: 598-604. doi: 10.1016/j.arabjc.2013.02.007.

Tsai S-J, Huang C-S, Mong M-C, Kam W-Y, Huang H-Y, Yin M-CJ (2012) Anti-inflammatory and antifibrotic effects of naringenin in diabetic mice. Agr Food Chem 60: 514–521. doi: 10.1021/jf203259h.

Ultee A, Bennik MHJ, Moezelaar R (2002) The phenolic hydroxyl group of carvacrol is essential for action against the food-borne pathogen Bacillus cereus. Appl Environ Microbiol 68: 1561-1568. doi: 10.1128/AEM.68.4.1561-1568.2002.

Wong SL, Chang HS, Wang GJ, Chiang MY, Huang HY, Chen CH, Tsai SC, Lin CH, Chen IS (2011) Secondary metabolites from the roots of Neolitsea daibuensis and their anti-inflammatory activity. J. Nat Prod 74: 2489-2496. doi: 10.1021/np100874f.