Journal of Chromatography A, 1217 (2010) 741–748 Contents lists available at ScienceDirect Journal of Chromatography A journal homepage: www.elsevier.com/locate/chroma Gas chromatographic–mass spectrometric analysis of volatiles obtained by four different techniques from Salvia rosifolia Sm., and evaluation for biological activity Gulmira Özek a,∗ , Fatih Demirci a , Temel Özek a , Nurhayat Tabanca b , David E. Wedge b , Shabana I. Khan c , Kemal Hüsnü Can Bas ¸ er a , Ahmet Duran d , Ergin Hamzaoglu e a Department of Pharmacognosy, Faculty of Pharmacy, Anadolu University, 26470 Eskisehir, Turkey b USDA, ARS, NPURU, National Center for Natural Products Research, The University of Mississippi, Mississippi, MS 38677, USA c National Center for Natural Products Research, Research Institute of Pharmaceutical Sciences, School of Pharmacy, The University of Mississippi, Mississippi, MS 38677, USA d Department of Biology, Faculty of Education, Selcuk University, 42090 Konya, Turkey e Department of Biology, Faculty of Science and Letters, Bozok University, Yozgat, Turkey article info Article history: Received 19 June 2009 Received in revised form 9 October 2009 Accepted 27 November 2009 Available online 2 December 2009 Keywords: Salvia rosifolia Essential oil Hydrodistillation Microwave Microdistillation Solid-phase microextraction -Pinene 1,8-Cineole Biological activity abstract Four different isolation techniques, conventional hydrodistillation (HD), microwave-assisted hydrodis- tillation (MWHD), microdistillation (MD) and micro-steam distillation-solid-phase microextraction (MSD-SPME), have been used to analyze the volatile constituents from the aerial parts of Salvia rosifolia Sm. by gas chromatography and gas chromatography coupled to mass spectrometry. HD and MWHD techniques produced quantitatively (yield, 0.39% and 0.40%) and qualitatively (aromatic profile) similar essential oils. -Pinene (15.7–34.8%), 1,8-cineole (16.6–25.1%), -pinene (6.7–13.5%), -caryophyllene (1.4–5.0%) and caryophyllene oxide (1.4–4.4%) were identified as major constituents of this Turkish endemic species. Besides, the hydrodistilled oil of S. rosifolia was evaluated for antibacterial, antifungal, anticancer, antioxidant and cytotoxic activities. The hydrodistilled oil of S. rosifolia showed antibacte- rial activity against Methicillin-resistant Staphylococcus aureus (MRSA) with a MIC value of 125 g/mL. Other human pathogenic microorganisms (Escherichia coli, Pseudomonas aeruginosa, Enterobacter aero- genes, Salmonella typhimurium, Staphylococcus epidermidis, Candida albicans) were also inhibited within a moderate range (MIC = 125–1000 g/mL). Antifungal activity of the oil was also observed against the strawberry anthracnose-causing fungal plant pathogens Colletotrichum acutatum, C. fragariae and C. gloeosporioides. No cytotoxicity was observed for S. rosifolia oil up to 25 mg/mL against malignant melanoma, epidermal, ductal and ovary carcinoma. © 2009 Elsevier B.V. All rights reserved. 1. Introduction The genus Salvia L. (Lamiaceae) encompasses over 900 species worldwide [1,2]. In Turkey Salvia is represented by 90 species, containing distinct 95 taxa, 47 of which are endemic. Since the last revision of the genus, four new species have been described from Turkey: Salvia nydeggeri Hub.-Mor. [3], Salvia aytachii Vural & Adiguzel [4], Salvia hedgeana Donmez [5], and Salvia anatolica Hamzaoglu & A. Duran [6]. Salvia is commonly known as “sage” in the world markets and represents one of the most diversified genera in Turkey with 52% endemic. Chemical diversity among Salvia taxa in Turkey has been reported by Bas ¸ er [7]. Several Salvia species are known locally as “adacayi” where they grow in southern and western Turkey and are ∗ Corresponding author. Tel.: +90 222 3350580/3719; fax: +90 222 3306809. E-mail address: gozek@anadolu.edu.tr (G. Özek). consumed as hot teas due to their unique flavor, pleasant aroma, and medicinal properties as well as are sources of essential oil important in cosmetics, perfumes, and medicine industries [8–11]. Antispasmodic, antibacterial, antifungal and antioxidant activities are reported for many Salvia species [12–15]. Several Turkish Salvia species have been reported earlier for the chemical constituents of their oils [16–21]. In terms of S. rosifolia there was published just one paper about its morphological and autecological properties [22]. Salvia plants and their essential oils are of economical impor- tance worldwide in food, pharmacy, perfumery, and cosmetics. So, investigation for applicability of different techniques for the isolation of Salvia oils with high yield and quality is very desir- able. The most common method for essential oil isolation is by hydrodistillation as described in various pharmacopoeias such as European Pharmacopoeia 2005 [23]. However, this technique has been controversial for subsequent determination of the oil chemical composition because of the possible transformation of 0021-9673/$ – see front matter © 2009 Elsevier B.V. All rights reserved. doi:10.1016/j.chroma.2009.11.086