Comparison of different extraction solvents for assay of the polyphenol content in the peel and pulp of 21 apple cultivars from Macedonia


  • Ana Petkovska Research and Development, AD Alkaloid, Skopje
  • Viktor Gjamovski Institute of Agriculture, Ss. Cyril and Methodius University
  • Marina Stefova Institute of Chemistry, Faculty of Natural Sciences and Mathematics, Ss. Cyril & Methodius University, Skopje



apples, polyphenols, flavonoids, spectrophotometry, extraction


Apples (Malus domestica Borkh.) contain a large array of phenolic compounds belonging to flavonoids and non-flavonoids. This study systematically evaluates the polyphenolic content of 21 apple cultivars from the Republic of Macedonia, both commercial and autochthonous, applying spectrophotometric methods for the determination of total polyphenolic compounds, total flavonoids, total anthocyanins and total flavan-3-ols. The reliability of these methods was checked by confirming the method linearity and accuracy with standards and spiked samples. The efficiency of acetone, water, methanol and a mixture of methanol/water (90 : 10, V/V) as extraction solvents was compared. Evident differences between extracts obtained from freeze-dried apple peel and pulp, extracted with different solvents, and analyzed with the four spectrophotometric tests, were observed and discussed. The most satisfactory extraction efficiency was achieved using methanol/water (90 : 10, V/V) mixture. The results obtained from the methanol/water extracts illustrate evident differences between polyphenol contents and reveal the diversity in polyphenols, total flavonoid and flavanol composition between the assayed commercial and autochthonous cultivars. The maximum abundance of polyphenols was noticed in the peel of Tetovka and Livadarka cultivars (15.63 mg/g GAE and 14.85 mg/g GAE, respectively) and in the pulp of Pasalma, Tetovka and Livadarka cultivars (12.55 mg/g GAE, 11.45 mg/g GAE and 11.22 mg/g GAE, respectively).


J. Janick, J. N. Cummins, S. K. Brown, and M. Hemmat, Apples: Fruit Breeding, Volume I: Tree and Tropical Fruits, John Wiley & Sons, Inc. 1996, pp. 9.

M. Cvetkovic, L. Tomic, M. Botu, V. Gjamovski, T. Jermic, B. Lazovic, V. Ognjanov, M. Pintea, R. Sevo, G. Achim, D. Bozovic, V. Bucarcuk, F. Carka, D. Cicek, G. Fruk, V. Jacimovic, M. Kiprijanovski, I. Hajlmarsson, Balkan Pomology Apples, Seednet Coordination, May 2012.

W. Oleszek, C. Y. Lee, A. W. Jaworski, K. R. Price, Identification of some phenolic compounds in apples, J. Agric. Food. Chem. 36, 430-432 (1988).

DOI: 10.1021/jf00081a007.

J. Pefes-Iizarbe, T. Hernandez, I. Estrella, Phenolic compounds in apples: varietal difference, Z. Lebensm. Unters. Forsch. 192(6), 551–554 (1991).

H. M. Merken, G. R. Beecher, Measurement of food flavonoids by high performance liquid chromatography: A review. J. Agric. Food Chem. 48, 577–599 (2000). DOI: 10.1021/jf990872o.

R. Tsao, R. Yang, J. C. Young, H. Zhu, Polyphenolic profiles in eight apple cultivars using High Performance Liquid Chromatography (HPLC), J. Agric. Food Chem. 51, 6347–6353 (2003). DOI: 10.1021/jf0346298.

J. J. Juan, R. Rodriguez, B. Suarez, A. Picinelli, E. Dapena, Study of the phenolic profile of cider apple cultivars at maturity by multivariate techniques, J. Agric. Food Chem. 47, 4046–4052 (1999).

DOI: 10.1021/jf9903197.

A. Escarpa, M. C. Gonzalez, High-performance liquid chromatography with diode array detection for the de-termination of phenolic compounds in peel and pulp from different apple varieties, J. Chromatogr. A, 823, 331–337 (1998). DOI: 10.1016/S0021-9673(98)00294-5.

E. Haslam, Y. Cai, Plant polyphenols (vegetable tan-nins): Gallic acid metabolism, Natural Product Reports 11(1), 41–66 (1994). DOI: 10.1039/NP9941100041.

S. P. Quideau, D. Deffieux, C. L. Douat-Casassus, L. Pouységu, Plant polyphenols: Chemical properties, biological activities, and synthesis, Angew. Chem. Int. Ed. 50(3), 586–621 (2011). DOI: 10.1002/anie.201000044.

D. K. Asami, Y. J. Hong, D. M. Barrett, A. E. Mitchell, Comparison of the total phenolic and ascorbic acid content of freeze-dried and air-dried marionberry, strawberry, and corn grown using conventional, organic, and sustainable agricultural practices. J. Agric. Food Chem. 51, 1237-1241 (2003). DOI: 10.1021/jf020635c.

Food and Agriculture Organization of the United Na-tions, Statistics Division (accessed in December 2015):,

V. Ivanova, M. Stefova, F. Chinnichi, Determination of polyphenol contents in Macedonian grapes and wines assessed by standardized spectrophotometric methods, J. Serb. Chem. Soc. 75(1), 45–59 (2010).

DOI: 10.2298/JSC1001045I.

J. Zhishen, T. Mengeheng, W. Jianming, The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals, Food Chem. 64, 555–559 (1999). DOI: 10.1016/S0308-8146(98)00102-2.

R. Di Stefano, M. C. Cravero, I composti fenolici e la natura del colore dei vini rossi, L’enotecnico Ottobre, 81 (1989).

R. Di Stefano, M. C. Cravero, N. Gentilini, Metodi per lo studio dei polifenoli dei vini, L’Enotecnico I Maggio, 83 (1989).

M. Kajdžanoska, J. Petreska, M. Stefova, Comparison of different extraction solvents mixtures for characterization of phenolic compounds in strawberries, J. Agric. Food Chem, 59, 5272–5278 (2011).

DOI: 10.1021/jf2007826.

M. Kajdžanoska, V. Gjamovski, M. Stefova, HPLC-DAD-ESI-MSn Identification of phenolic compounds in cultivated strawberries from Macedonia, Maced. J. Chem. Chem. Eng 29(2), 181–194 (2010).

D. Marinova, F. Ribarova, M. Atanassova, Total Phenolics and total flavonoids in Bulgarian fruits and vegetables, J. Univ. Chem. Tech. Metall. 40(3), 255–260 (2005).

M. Ceymann, E. Arrigoni, H. Schärer, D. Baumgartner, A. Bozzi Nising, R.F. Hurrell, Rapid high performance screening method using UHPLC-MS to quantify 12 polyphenol compounds in fresh apples, Anal. Methods 3, 1774–1778 (2011). DOI: 10.1039/c1ay05152k.

S. Guyot, C.L. Bourvellec, N. Marnet, J. F. Drilleau, Procyanidins are the most abundant polyphenols in dessert apples at maturity, LWT–Food Sci. Technol. 35, 289–291 (2002). DOI: 10.1006/fstl.2001.0843.

U. Vrhovsek, A. Rigo, D. Tonon, F. Mattivi, Quantita-tion of polyphenols in different apple varieties, J. Agric. Food Chem.52, 6532–6538 (2004).

DOI: 10.1021/jf049317z.


Additional Files



How to Cite

Petkovska, A., Gjamovski, V., & Stefova, M. (2016). Comparison of different extraction solvents for assay of the polyphenol content in the peel and pulp of 21 apple cultivars from Macedonia. Macedonian Journal of Chemistry and Chemical Engineering, 35(1), 29–38.



Food Chemistry

Most read articles by the same author(s)

> >>