Analysis of Wild Raspberries (Rubus idaeus L.): Optimization of the Ultrasonic-Assisted Extraction of Phenolics and a New Insight in Phenolics Bioaccessibility

  • Nevena R. Mihailović
  • Vladimir B. Mihailović
  • Andrija R. Ćirić
  • Nikola Z. Srećković
  • Mirjana R. Cvijović
  • Ljubinka G. JoksovićEmail author
Original Paper


A simple and efficient ultrasonic-assisted extraction (UAE) technique was developed in order to find optimal conditions for the extraction of total phenolic compounds, flavonoids and anthocyanins in wild raspberry (Rubus idaeus L.) fruits. Several extraction variables, including methanol composition (v/v, %), solid-solvent ratio (g/mL), time (min) and extraction temperature (°C) were optimized using response surface methodology (RSM). Under optimal conditions for extraction, the total phenolics were found in the concentration of 383 mg GAE/100 g of fresh fruit weight, while HPLC-PDA analysis of the optimized extract showed the presence of cyanidin-3-glucoside, cyanidin-3-sophoroside, catechin, gallic and ellagic acid. The experimental values of DPPH and ABTS radical scavenging activities were 29.0 and 39.5 μmol Trolox/g of fresh fruit weight, respectively. In vitro simulated gastrointestinal digestion showed great raspberry phenolics stability. Our study assessed the bioaccessible phenolics in wild raspberry fruits and showed optimal conditions for the effective extraction of bioactive compounds for their analysis.


Rubus idaeus L. Extraction HPLC-PDA Cyanidin-3-glucoside Antioxidant activity In vitro digestion 



2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)


Central composite design


Cyanidin-3-glucoside equivalents




Fresh weight


Gallic acid equivalents


High performance liquid chromatography


Photo diode array


Retention time


Response surface methodology


Rutin equivalents


Total anthocyanins content


Total flavonoids content


Total phenolics content


Ultrasonic-assisted extraction


Ultra violet




Wild raspberry fruit



This work is supported by the Ministry of Education, Science and Technological Development of the Republic of Serbia (Grants no. OI 172016, III 43004 and OI 175039).

Compliance with Ethical Standards

Conflict of Interest

The authors declare no conflict of interest.

Human and Animal Studies

This article does not contain any studies with human or animal subjects.

Informed Consent

Informed consent was obtained from all individual participants included in the study.

Supplementary material

11130_2019_756_MOESM1_ESM.docx (924 kb)
ESM 1 (DOCX 923 kb)


  1. 1.
    Dantas AM, Mafaldo IM, Oliveira PM, Lima MDS, Magnani M, Borgess GDSC (2019) Bioaccessibility of phenolic compounds in native and exotic frozen pulps explored in Brazil using a digestion model coupled with a simulated intestinal barrier. Food Chem 274:202–214CrossRefGoogle Scholar
  2. 2.
    Rao AV, Snyder DM (2010) Raspberries and human health: a review. J Agric Food Chem 58:3871–3883CrossRefGoogle Scholar
  3. 3.
    Snyder SM, Low RM, Stocks JC, Eggett DL, Trker TL (2012) Juice, pulp and seeds fractionated from dry climate Primocane raspberry cultivars (Rubus idaeus) have significantly different antioxidant capacity, anthocyanin content and color. Plant Foods Hum Nutr 67:358–364CrossRefGoogle Scholar
  4. 4.
    Rauha JP, Remes S, Heinonen M, Hopia A, Kähkönen M, Kujala T, Pihlaja K, Vuorela H, Vuorela P (2000) Antimicrobial effects of Finnish plant extracts containing flavonoids and other phenolic compounds. Int J Food Microbiol 56:3–12CrossRefGoogle Scholar
  5. 5.
    Baby B, Antony P, Vijayan R (2018) Antioxidant and anticancer properties of berries. Crit Rev Food Sci 58:2491–2507CrossRefGoogle Scholar
  6. 6.
    Määttä-Riihinen KR, Kamal-Eldin A, Törrönen AR (2014) Identification and quantification of phenolic compounds in berries of Fragaria and Rubus species (family Rosaceae). J Agric Food Chem 52:6178–6187CrossRefGoogle Scholar
  7. 7.
    Tosun M, Ercisli S, Karlidag H, Sengul M (2009) Characterization of red raspberry (Rubus idaeus L.) genotypes for their phytochemical properties. J Food Sci 74:C575–C579CrossRefGoogle Scholar
  8. 8.
    Zadernowski R, Naczk M, Nesterowicz J (2005) Phenolic acid profiles in some small berries. J Agric Food Chem 53:2118–2124CrossRefGoogle Scholar
  9. 9.
    Tao Y, Wu D, Zhang QA, Sun DW (2014) Ultrasound-assisted extraction of phenolics from wine lees: modeling, optimization and stability of extracts during storage. Ultrason Sonochem 21:706–715CrossRefGoogle Scholar
  10. 10.
    Carrera C, Ruiz-Rodriguez A, Palma M, Barroso CG (2012) Ultrasound assisted extraction of phenolic compounds from grapes. Anal Chim Acta 732:100–104CrossRefGoogle Scholar
  11. 11.
    Ciğeroğlu Z, Aras Ö, Pinto CA, Bayramoglu M, Kırbaşlar Şİ, Lorenzo JM, Barba FJ, Saraiva JA, Şahin S (2018) Optimization of ultrasound-assisted extraction of phenolic compounds from grapefruit (Citrus paradisi Macf.) leaves via D-optimal design and artificial neural network design with categorical and quantitative variables. J Sci Food Agric 98:4584–4596CrossRefGoogle Scholar
  12. 12.
    Korkut I, Bayramoglu M (2014) Various aspects of ultrasound assisted emulsion polymerization process. Ultrason Sonochem 21:1592–1599CrossRefGoogle Scholar
  13. 13.
    Tavčar Benković E, Grohar T, Žigon D, Švajger U, Janeš D, Kreft S, Štrukelj B (2014) Chemical composition of the silver fir (Abies alba) bark extract Abigenol® and its antioxidant activity. Ind Crop Prod 52:23–28CrossRefGoogle Scholar
  14. 14.
    Singleton VL, Orthofer R, Lamuela-Raventós RM (1999) Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. In: Lester P (ed) Methods in enzymology. Academic Press, Cambridge, pp 152–178Google Scholar
  15. 15.
    Brighente IMC, Dias M, Verdi LG, Pizzolatti MG (2007) Antioxidant activity and total phenolic content of some Brazilian species. Pharm Biol 45:156–161CrossRefGoogle Scholar
  16. 16.
    Giusti MM, Wrolstad RE (2001) Anthocyanins. Characterization and measurement of anthocyanins by UV-visible spectroscopy. In: Wrolstad RE (ed) Current protocos in food analytical chemistry. John Wiley & Sons, New York unit F1.2.1−1Google Scholar
  17. 17.
    Kumarasamy Y, Byres M, Cox PJ, Jaspars M, Nahar L, Sarker SD (2007) Screening seeds of some Scottish plants for free-radical scavenging activity. Phytother Res 21:615–621CrossRefGoogle Scholar
  18. 18.
    Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, RiceEvans C (1999) Antioxidant activity applying an improved ABTS radical cation decolorizationassay. Free Radic Biol Med 26:1231–1237CrossRefGoogle Scholar
  19. 19.
    Minekus M, Alminger M, Alvito P, Ballance S, Bohn T, Bourlieu C, Carriére F, Boutrou R et al (2014) A standardised static in vitro digestion method suitable for food– an international consensus. Food Funct 5:1113–1124CrossRefGoogle Scholar
  20. 20.
    Mihailović V, Kreft S, Tavčar Benković E, Ivanović N, Stanković MS (2016) Chemical profile, antioxidant activity and stability in stimulated gastrointestinal tract model system of three Verbascum species. Ind Crop Prod 89:141–151CrossRefGoogle Scholar
  21. 21.
    Robards K (2003) Strategies for the determination of bioactive phenols in plants, fruit and vegetables. J Chromatogr A 1000:657–691CrossRefGoogle Scholar
  22. 22.
    Derringer G, Suich R (1980) Simultaneous optimization of several response variables. J Qual Technol 12:214–219CrossRefGoogle Scholar
  23. 23.
    Milivojevic J, Maksimovic V, Nikolic M, bogdanovic J, Maletic R, Milatovic D (2011) Chemical and antioxidant properties of cultivated and wild Fragaria and Rubus berries. J Food Qual 34:1–9CrossRefGoogle Scholar
  24. 24.
    Yang JW, Choi IS (2017) Comparison of the phenolic composition and antioxidant activity of Korean black raspberry, Bokbunja, (Rubus coreanus Miquel) with those of six other berries. CyTA – J Food 15:110–117. Google Scholar
  25. 25.
    Diaconeasa Z, Ranga F, Rugină D, Leopold L, Pop O, Vodnar D, Cuibus L, Socaciu C (2015) Phenolic content and their antioxidant activity in various berries cultivated in Romania. Bull UASVM Food Sci Technol 72:99–103Google Scholar
  26. 26.
    Mikulic-Petkovsek M, Schmitzer V, Slatnar A, Stampar F, Veberic R (2012) Composition of sugars, organic acids, and total phenolics in 25 wild or cultivated berry species. J Food Sci 77:C1064–C1070CrossRefGoogle Scholar
  27. 27.
    Bobinaite R, Viškelis P, Venskutonis PR (2012) Variation of total phenolics, anthocyanins, ellagic acid and radical scavenging capacity in various raspberry (Rubus spp.) cultivars. Food Chem 132:1495–1501CrossRefGoogle Scholar
  28. 28.
    Sariburun E, Sahin S, Demir C, Turkben C, Uylaser V (2010) Phenolic content and antioxidant activity of raspberry and blackberry cultivars. J Food Sci 75:C328–C335CrossRefGoogle Scholar
  29. 29.
    Strugala P, Loi S, Bazanow B, Kuropka P, Kucharska AZ, Wloch A, Gabrielska J (2018) A comprehensive study on the biological activity of elderberry extract and cyanidin 3-O-glucoside and their interactions with membranes and human serum albumin. Molecules 23:E2566. CrossRefGoogle Scholar
  30. 30.
    de Souza VR, Pereira PA, da Silva TL, de Oliveira Lima LC, Pio R, Queiroz F (2014) Determination of the bioactive compounds, antioxidant activity and chemical composition of Brazilian blackberry, red raspberry, strawberry, blueberry and sweet cherry fruits. Food Chem 156:362–368CrossRefGoogle Scholar
  31. 31.
    McDougall GJ, Dobson P, Smith P, Blake A, Stewart D (2005) Assessing potential bioavailability of raspberry anthocyanins using an in vitro digestion system. J Agric Food Chem 53:5896–5904CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Faculty of Science, Department of ChemistryUniversity of KragujevacKragujevacSerbia
  2. 2.Faculty of Medicine, Institute for Microbiology and ImmunologyUniversity of BelgradeBelgradeSerbia

Personalised recommendations