Contents lists available at ScienceDirect Journal of Archaeological Science: Reports journal homepage: www.elsevier.com/locate/jasrep Evidence of tobacco from a Late Archaic smoking tube recovered from the Flint River site in southeastern North America S. Carmody a , J. Davis b , S. Tadi c , J.S. Sharp c , R.K. Hunt d , J. Russ d, ⁎ a Department of Social Sciences, Troy University, 132D MSCX, Troy, AL 36082, USA b New South Associates, Inc., 6150 East Ponce de Leon Avenue, Stone Mountain, GA 30083, USA c Department of BioMolecular Sciences, School of Pharmacy, University of Mississippi, 72 University Avenue, University, MS 38677, USA d Department of Chemistry, Rhodes College, 2000 North Parkway, Memphis, TN 38112, USA ARTICLE INFO Keywords: Tobacco Nicotine Gas chromatography-mass spectrometry smoking pipes organic residue analysis ABSTRACT Tobacco use was widespread amongst the indigenous populations throughout North and South America prior to European contact; however, the geographical and temporal spread of the plant is poorly understood. Organic residue analysis is providing a new source of information on the diffusion of tobacco based on the presence of nicotine extracted from smoking pipes recovered from archaeological contexts. Using gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-high-resolution mass spectrometry (LC-HRMS) we identified nicotine in a Late Archaic (1685-1530 cal B.C) smoking pipe from the Flint River site in Northern Alabama. This evidence suggests the exploitation of tobacco spread into the southeastern North America nearly a millennium earlier than the current hypothesis on the rate of dispersion of tobacco in pre-Columbian North America. 1. Introduction Tobacco was omnipresent in the New World prior to European contact. Most Native American communities revered tobacco as sacred, assigning it a central role in many religious ceremonies (Winter, 2000a). The ancestral source of Nicotiana rustica and Nicotiana tabacum, the first tobacco species humans exploited, has been traced genetically to the northern Andean Highlands in South America (Goodspeed, 1954; Aoki and Ito, 2000; Clarkson et al., 2017). But the geographical spread and the pace of tobacco's expansion throughout the New World, and how and why it came to be a primary focus of religious practices, re- mains poorly understood. Current hypotheses on the diffusion of tobacco are based largely on archaeobotanical evidence (Wagner, 2000). Direct evidence of tobacco plants, principally seeds, has been observed at more than 100 archae- ological sites in North America, most dating to between 1000 and 1600 CE, with the oldest deposited between 80 and 240 CE (Asch, 1994; Winter, 2000b). While the search for botanical evidence has proven successful, it is limited by the paucity of tobacco pollen in archae- ological contexts, especially at sites that are more than one thousand years old. Chemical residue analysis of pipe contents has emerged as an al- ternative method for addressing tobacco use in prehistory (Tushingham and Eerkens, 2016). Smoking pipes are prevalent in the North American archaeological record, particularly in eastern North America where most pipes are associated with human burials (Rafferty, 2016). Tobacco use can be deduced by the presence of nicotine – the primary alkaloid in most Nicotiana species – in pipe residues or within the artifact matrix. Early work by Gager et al. (1960) identified nicotine in a 7th century pipe, demonstrating that the alkaloid can persist for more than a thousand years. More recently, Rafferty (2002, 2006; Rafferty et al., 2012) detected nicotine in a variety of smoking pipes collected from sites in the American Northeast. Likewise, organic residue analysis has identified nicotine at sites in the Pacific Northwest (Eerkens et al., 2012; Tushingham et al., 2013), including in two well-dated smoking tubes discovered in graves dating to the early 15th century. In the Midwest, Freimuth et al. (2012) detected nicotine in one of three pipes recovered from late 8th century contexts at a site in Eastern Missouri. Organic residue analysis has pushed back the time for the in- troduction of tobacco in North America, with the earliest evidence coming from a smoking tube dated at between 500 and 300 years BCE by Rafferty et al. (2012). There remains, however, a substantial time gap between the oldest archaeologically recovered pipes, which enter the archaeological record between 3000 and 1000 BCE (Lewis and Lewis, 1961), and the earliest direct evidence for tobacco use. This time gap indicates that either tobacco was added to a pre-existing and an- cient smoking complex, or simply that our methods have yet to detect evidence of much earlier tobacco use. Either way, this time gap https://doi.org/10.1016/j.jasrep.2018.05.013 Received 11 March 2018; Received in revised form 21 May 2018; Accepted 25 May 2018 ⁎ Corresponding author. E-mail address: russj@rhodes.edu (J. Russ). Journal of Archaeological Science: Reports 21 (2018) 904–910 Available online 15 June 2018 2352-409X/ © 2018 Elsevier Ltd. All rights reserved. T