pubs.acs.org/JAFC Published on Web 05/18/2009 © 2009 American Chemical Society J. Agric. Food Chem. 2009, 57, 4663–4667 4663 DOI:10.1021/jf9005092 Chemical and Genetic Relationships among Sage (Salvia officinalis L.) Cultivars and Judean Sage (Salvia judaica Boiss.) ANDREA BO ¨ SZO ¨ RME ´ NYI,* ,†  EVA HE ´ THELYI, †  AGNES FARKAS, § GYO ¨ RGYI HORVA ´ TH, § NO ´ RA PAPP, §  EVA LEMBERKOVICS, † AND  EVA SZ 00 OKE † † Faculty of Pharmacy, Institute of Pharmacognosy, Semmelweis University, :: Ull 00 Oi  ut. 26, 1085 Budapest, Hungary, and § Medical School, Institute of Pharmacognosy, University of Pe´cs, Ro´ kus u. 2, 7624 Pe´cs, Hungary The essential oil composition and genetic variability of common sage (Salvia officinalis L.) and its three ornamental cultivars (‘Purpurascens’, ‘Tricolor’, and ‘Kew Gold’) as well as Judean sage (Salvia judaica Boiss.) were analyzed by GC-FID, GC-MS, and random amplified polymorphic DNA (RAPD). Common sage and its cultivars contained the same volatile compounds; only the ratio of compounds differed. The main compounds were the sesquiterpene R-humulene and the mono- terpenes β-pinene, eucalyptol, and camphor. Judean sage contained mainly the sesquiterpenes β-cubebene and ledol. All of the samples exhibited characteristic RAPD patterns that allowed their identification. Cluster analyses based on oil composition and RAPD markers corresponded very well to each other, suggesting that there is a strong relationship between the chemical profile and the genetic variability. KEYWORDS: Essential oil; gas chromatography; mass spectroscopy; RAPD; genetic distances; cluster analysis INTRODUCTION Common sage (Salvia officinalis L., Lamiaceae) is one of the most important medicinal plants in the mint family. Sages are frequent components in several multiherb products and food supplements such as teas, tinctures, liquid extracts, rheuma patches, and cosmetics (1, 2). Sage is commonly used also as a culinary herb, listed by the Council of Europe (3 ) as a natu- ral source of food flavoring. It can be added to foodstuffs providing that the concentration of thujones present in the final product does not exceed 0.5 mg/kg. In the United States sage is listed as “generally recognized as safe” (GRAS) (4 ). Its drug and tincture are official in several Pharmacopoeias (e.g., Pharmacopoeia Hungarica, 8th ed.; European Pharmacopoeia, 6th ed.) (5, 6). The plant is reported to have a wide range of biological activities, such as antibacterial, fungistatic, virustatic, estrogenic, anticholinesterase, adstringent, eupeptic, and antihydrotic effects (7, 8). The antimicrobial properties as well as the tannin-based astringent activities of sage (active ingredient of dental-care herbal medicinal preparations) reduce plaque growth, inhibit gingival inflammation, and have positive effects on caries prophylaxis (9 ). The leaves of sage are well-known for their antioxidative properties; other experimental studies on sage extracts or sage essential oil showed hypotensive, central ner- vous system-depressant actions, and antispasmodic activities. Furthermore, sage is externally used for the treatment of insect bites (10, 11). The strongest active constituents of sage are within its essential oil (1-2.8%), comprising the monoterpenes R- and β-thujone, camphor, cineole, and borneol as well as the sesquiterpenes R-humulene and β-caryophyllene in larger amounts, whereas the leaf contains di- and triterpenes, as well. Due to the presence of toxic thujone components, sage oil should be used with caution. Other constituents of the herb that contribute to its antioxidant effect are phenolic acids (caffeic, chlorogenic, ros- marinic, ferulic), flavonoids, and tannins (12-14). The chemical composition of sage essential oil has been investigated in various countries (15, 16), and the essential oils were divided into five groups according to the amount of the major constituents: 1. camphor > R-thujone >1,8-cineole > β-thujone 2. camphor > R-thujone > β-thujone > 1,8-cineole 3. β-thujone > camphor > 1,8-cineole > R-thujone 4. 1,8-cineole > camphor > R-thujone > β-thujone 5. R-thujone > camphor > β-thujone > 1,8-cineole Further authors mentioned viridiflorol (sesquiterpene-alcohol) as one of the five major components (17 ). ISO standard 9909:1997 for the essential oil composition of common sage prescribes the following: R-thujone, 18.0-43.0%; β-thujone, 3.0-8.5%; camphor, 4.5-24.5%; 1,8-cineole, 5.5-13.0%; R-humulene, 0-12%; R-pinene, 1.0-6.5%; camphene, 1.5- 7.0%; limonene, 0.5-3.0%; linalool and its esters, <1%; and bornyl acetate, <2.5% (18 ). Variation in essential oil yield and constitution can be due to both environmental and genetic factors. Echeverrigaray et al. (19 ) found, for example, that the population level diversity in essential oil composition of dittany (Cunila galioides Benth.) is *Author to whom correspondence should be addressed (e-mail aboszormenyi@gmail.com; telephone/fax 3613172979).