Rapid Species-level Identification of Salvias by Chemometric Processing of Ambient Ionisation Mass Spectrometry-derived Chemical Profiles Justine E. Giffen, a Ashton D. Lesiak, a A. John Dane, b Robert B. Cody b and Rabi A. Musah a * ABSTRACT: Introduction – The Salvia genus contains numerous economically important plants that have horticultural, culinary and nutraceu- tical uses. They are often similar in appearance, making species determination difficult. Species identification of dried Salvia products is also challenging since distinguishing plant morphological features are no longer present. Objective – The development of a simple high-throughput method of analysis of fresh and dried Salvia leaves that would permit rapid species-level identification and detection of diagnostic biomarkers. Methodology – Plant leaves were analysed in their native form by DART-MS without the need for any sample preparation steps. This furnished chemical fingerprints characteristic of each species. In the same experiment, in-source collision-induced dissocia- tion was used to identify biomarkers. Biomarker presence was also independently confirmed by GC–MS. Chemometric processing of DART-MS profiles was performed by kernel discriminant analysis (KDA) and soft independent modelling of class analogy (SIMCA) to classify the fingerprints according to species. Results – The approach was successful despite the occurrence of diurnal cycle and plant-age related chemical profile variations within species. In a single rapid experiment, the presence of essential oil biomarkers such as 3-carene, α-pinene, β-pinene, β-thujone, β-caryophyllene, camphor and borneol could be confirmed. The method was applied to rapid identification and differ- entiation of Salvia apiana, S. dominica, S. elegans, S. officinalis, S. farinacea and S. patens. Conclusion – Species-level identification of Salvia plant material could be accomplished by chemometric processing of DART- HRMS-derived chemical profiles of both fresh and dried Salvia material. Copyright © 2016 John Wiley & Sons, Ltd. Supporting information can be found in the online version of this article. Keywords: DART-MS; ambient ionisation; sage; Salvia; multivariate statistical analysis; triterpenoids; essential oils; SIMCA; KDA Introduction The Salvia genus, commonly referred to as Sage, is one of the most agriculturally important taxa in the world due to the commercial importance of its plants in the culinary, medicinal, fragrance and horticultural sectors, and to its wide geographic distribution. Among the culinary sages are Salvia officinalis L., Salvia elegans Vahl (pineapple sage), S. fruticose (Greek sage) and S. melissordora ( grape scented sage). Ornamental sages include Salvia apiana Jeps. (white sage), S. discolor, S. greggii, Salvia farinacea Benth., S. microphylla and S. clary among numerous others. Salvia essential oils, particularly those from S. officinalis and S. lavandulaefolia, have been reported to exhibit memory enhancing effects (Eidi et al., 2006). This has been attributed to the synergistic impact of constit- uent terpenes and terpenoids such as α-pinene, geraniol, linalool, γ-terpinene, 1,8-cineole and camphor acting on cholinergic recep- tors (Miroddi et al., 2014). Salvia spp. plants have also been re- ported to exhibit antimicrobial ( Jirovetz et al., 2006; Jirovetz et al., 2007; Benkherara et al., 2015), anti-inflammatory (Moretti et al., 1997; Ehrnhöfer-Ressler et al., 2013), anticancer (Russo et al., 2013) and antioxidant activities (Kozics et al., 2013). As it is the essential oils of the plants which are important for many of its culinary and medicinal applications, significant attention has been given to determining the defining features of the essential oil profiles of Salvia species plants. In this regard, a number of trends have been observed. For example, although Salvias are often found to contain terpenes and terpene derivatives such as α- and β-pinene, camphene, 1,8-cineole, α- and β-thujone, cam- phor, borneol, α- and β-caryophyllene and viridiflorol (Oelschlägel et al., 2012), the relative proportions and appearance of these constituents in a particular plant part vary widely depending upon species. Thus, while the leaves of S. officinalis have consistently been found to contain the aforementioned terpenes, S. elegans leaves have been found to be devoid of β-pinene, 1,8-cineole, α- and β-thujone and camphor (Oelschlägel et al., 2012). It has also been shown that constituent composition can vary as a function of intraspecies genetic variability (Perry et al., 1999), plant part (Perry et al., 1999; Santos-Gomes and Fernandes-Ferreira, 2001), season (Putievsky et al., 1986; Müller-Riebau et al., 1997), flowering versus vegetative stage (Piccaglia et al., 1997) and harvest date (Perry et al., 1999). Despite these observations, it is also clear that * Correspondence to: Rabi A. Musah, Department of Chemistry, University at Al- bany, State University of New York, 1400 Washington Avenue, Albany, NY 12222, USA. Email: rmusah@albany.edu a Department of Chemistry, University at Albany, State University of New York, 1400 Washington Avenue, Albany, NY, 12222, USA b JEOL USA Inc., 11 Dearborn Road, Peabody, MA, 01960, USA Phytochem. Anal. 2017, 28, 16–26 Copyright © 2016 John Wiley & Sons, Ltd. Research Article Received: 20 March 2016, Revised: 25 July 2016, Accepted: 3 August 2016 Published online in Wiley Online Library (wileyonlinelibrary.com) DOI 10.1002/pca.2639 16