Selected bioactives from callus cultures of olives (Olea europaea L. Var. Coratina) by LC-MS Luigi Gentile a,b , Nicola A. Uccella c,d, ⁎ a Computer Science, Modelling, Electronics and Systems Engineering Department, University of Calabria, via P. Bucci, Rende, CS, Italy b Department of Chemistry and Chemical Technologies, University of Calabria, via P. Bucci 12C, 87036 Rende, CS, Italy c IRESMO Foundation Group, via Cavour 5, Montalto Uffugo, 87040 CS, Italy d Dipartimento di Ingegneria per l'Ambiente e il Territorio e Ingegneria Chimica, University of Calabria, via P. Bucci, Rende, CS, Italy abstract article info Article history: Received 5 July 2013 Accepted 27 October 2013 Keywords: Coratina olive callus culture Biophenol Secoiridoids HPLC–ESI-MS ESI-MS/MS-CAD Bond dissociation enthalpy Olive biophenols, OBPs, and biophenol-secoiridoids, OBPsecos, soluble bioactives from olives, and then in olive oil and table olives, were biosynthesized from Coratina olive callus cultures, under bio- and techno-mimetic exper- imental conditions. We detected major production of OBPs, hydroxytyrosol, tyrosol, caffeic acid and verbascoside, and of OBP-secos, oleuropein and ligstroside, together with minor amounts of oleuroside, and of demethyl and deglucosil bioactives, by high-performance liquid chromatography, HPLC, coupled with electrospray ionization mass spectrometry, ESI-MS. The olive callus system, in an experimental non-pathogenic environment, produced a nearly model balance of OBPs and OBPsecos in suitable amount comparable to drupes, leaves and seeds. The bioreactor scaling-up of more homogeneous and rapidly growing systems, processed by callus shaking to suspen- sion cell cultures, should produce nutraceutical and cosmeceutical bioactives. © 2013 Elsevier Ltd. All rights reserved. 1. Introduction Mediterranean olive products, olive oil and table olives (Charoenprasert & Mitchell, 2012; Estruch et al., 2013), belong to func- tional foods, for which beneficial effects (Agostoni et al., 2011; Frankel, 2011; Obied, 2013) have already been claimed for consumer health and well-being. Olive oil, the first factor among several others in ancient diet, as declared by UNESCO Mediterranean Diet, MD, as an Intangible Cultural Heritage of Humanity, is more than an ingredient in the careful selections of nutrients and weight guidelines (UNESCO, 2010), descend- ing from the Mediterranean Aliment Culture, MAC (Uccella, 2003). Health benefits of MAC traditional olive oil and table olives depend on peculiar phytochemicals of bio- and techno-derivation (Agostoni et al., 2011). Monoenoic fatty acids in triacylglycerols, TAGs, 98–99%, modulate blood pressure reduction with n–9 oleic acid (Teres et al., 2008), memory consolidation through oleoylethanolamide (Campolongo et al., 2009), cell membrane fluidity (Filippelli, Oliviero Rossi, & Uccella, 2011) and longevity (Puca et al., 2008) with n–7, asclepic and palmitoleic acids, respectively. Minor MAC bioactive phenols in olives are commonly called soluble olive biophenols (Obied, 2013), i.e., tyrosol, Ty, hydroxytyrosol, HTy, caffeic acid, CA, and verbascoside, Ve, Fig. 1, while further phytochemicals are soluble seco-complexes. Oleuropein, Ole, ligstroside, Lig, their C11- demethylated analogs, demethyloleuropein and demethylligstroside, are OBPs esterified to C7-carbonyl of 11-methyl oleoside and of iso-oleoside in oleuroside. Nuezhenide, Nue, has the glucopyranosil bridge between OBP and seco-group (De Leonardis, Macciola, & Nag, 2009). OBPs and OBPsecos, glucosilated at deoxyloganic acid level (Gutierrez-Rosales, Romero, Casanovas, Motilva, & Minguez-Mosquera, 2012), towards water-solubility, masked reactivity, ease vacuole storage and functional effects, are soluble, esterified, and insoluble cell-wall bound within cellular tissues from Oleaceae (Uccella, 2001a,b), Gentianaceae and Cornaceae families, genus Olea, Fraxinus, Jasminum, and Osmanthus (Estruch et al., 2013). OBPs and OBPsecos from leaves flow into olive oil during olive crushing, as in the MAC practice of adding leaves to fruit bulks. This old-fashioned process, inherited from MAC wisdom in local frantoi, olive mills, allows their mixing up and partitioning with triterpenoids as well (Allouche et al., 2008). Thus, by-products of the olive agro- industry (Herrero et al., 2011; Rubio, Motliva, Macia, Ramo, & Romero, 2012; Suarez, Romero, Ramo, Macia, & Motilva, 2008; Zbakh & El Abbassi, 2012) prevent spoilage of low free acidity olive oils against oxidation and hedonic-sensorial loss, and restore the beneficial effects of the end product, distinctive for its sensorial quality of a persistent bitter taste and intense green color (Frankel, 2011; Obied, 2013; Uccella, 2003; UNESCO, 2010). Chimeric forms of OBPsecos overcome major drawbacks of separate functionalities through complemental and strategic functions in olive defensive systems, human health and in effects on well-being (Charoenprasert & Mitchell, 2012; Uccella, Toscano, & Piperno, 2004). Ty and HTy exploit free radical scavenging, singlet oxygen quenching Food Research International 55 (2014) 128–136 ⁎ Corresponding author at: IRESMO Foundation Group, via Cavour 5, Montalto Uffugo, 87040 CS, Italy. Fax: +39 0984492116. E-mail address: nicola.uccella@unical.it (N.A. Uccella). 0963-9969/$ – see front matter © 2013 Elsevier Ltd. All rights reserved. http://dx.doi.org/10.1016/j.foodres.2013.10.046 Contents lists available at ScienceDirect Food Research International journal homepage: www.elsevier.com/locate/foodres