The Biomolecular Archaeology of the Black Drink: Alkaloid Residue Analysis of Ilex vomitoria on Experimental Vessels and Applications for Prehistoric Specimens Paper presented at the 2011 Southeastern Archaeological Conference, Jacksonville, Florida Shannon Tushingham1*, Matthew Palumbo2, Charles H. McNutt3, and Ulla Anderson4 1Department of Anthropology, Washington State University’ 2Department of Biology, University of Florida; 3Department of Anthropology, University of Memphis; 4Department of Chemistry, UC Berkeley; *Corresponding Author: [email protected] ABSTRACT Early historic accounts document widespread use of a purgative known as “the black drink”, a caffeinated ceremonial tea famous for its use in purification rituals by elite males. The beverage was prepared from roasted leaves and twigs of yaupon holly (Ilex vomitoria), an evergreen shrub native to the southeastern coastal plain. Prehistoric use is poorly understood but often associated with shell cups found in mortuary contexts and the SECC. Our work, designed to track ancient use of psychoactive plants via Gas Chromatography‐Mass Spectrometry, includes identification of medicinal tea biomarkers, experimentation with brewing black drink, and residue extraction from pottery/shell specimens. Sampling included preparing and extracting a range of experimental vessels which served different functions in black drink cooking and consumption (cooking vessels, clay and shell cups), with extractions taken directly from the vessels and from tea residue on vessel interiors. Our findings suggest that chemical changes occur through the cooking process, but the presence of the biomarker caffeine remains stable and correlated the experimental vessels to the Ilex vomitoria plant in all but one case. We also describe the non‐destructive method we developed where alkaloid residues can be extracted directly from the matrix of prehistoric shell cups and pottery vessels through a sonication process. INTRODUCTION Early historic accounts document widespread use of a purgative known as “the black drink”, a caffeinated ceremonial tea used as an everyday beverage by many southeastern Indians, but perhaps most famous for its use in dramatic purification rituals by elite males. The beverage was prepared from roasted leaves and twigs of Ilex vomitoria, a plant popularly known as the yaupon holly. The black drink seems to have been part of a religious or ceremonial system, reaching back perhaps thousands of years. As the black drink was consumed in Figure 1. 16th-century engraving by Jacques le Moyne of special marine shell cups that were often buried by high Timucan Black Drink Ceremony, Florida. Note seated status historic southeastern Indians, it is thought that individuals vomiting, women in right foreground prehistoric shell cups found in similar contexts, brewing black drink, and European observers in left foreground. Copy from Theodor de Bry (Frankfort: Typis particularly those engraved with Southeastern J. Wecheli, 1591). Smithsonian Institution National Ceremonial Complex iconographic symbols, may have Anthropological Archives negative no. 57,569. Tushingham et al. 2011‐2 served a similar function. This is, however, speculation, as we simply have no direct evidence of the medicinal teas that may or may not have been used in these vessels. In this paper we describe our research that is designed to track ancient use of the black drink and other ritual beverages via Gas Chromatography‐Mass Spectrometry. This work involves the identification of medicinal tea biomarkers, experimentation with brewing black drink, and residue extraction from pottery and shell specimens. We are specifically interested in understanding the chemistry of these plants, looking at how this chemistry might change through the cooking process, and how we might expect residues with identifiable alkaloid residues to preserve archaeologically. Our greater intent is to establish non‐destructive extraction methods that can be used on prehistoric specimens. HISTORIC NATIVE AMERICAN USE There are ample historic accounts of Native American use of the black drink dating to as early as the 16th century (see Hudson 1979, Merrill 1979 and Fairbanks 1979 for summaries of this material). Perhaps the most famous image of a black drink ceremonial is shown in an engraving from the 1560s by a member of a Florida expedition where explorers recorded observations of local native people (Figure 1). In this illustration we see a chief and his advisors drinking the black drink in shell cups during a meeting. Note the men profusely regurgitating the beverage at their feet, the women preparing the drink in large pots in the lower right, and the European explorers observing the spectacle in the lower left. From the ethnography, it is clear that the black drink was connected to purity, which found its most dramatic expression in those contexts where it was regurgitated. The beverage was consistently restricted to certain individuals, in particular men of a certain age or status. Though used to induce vomiting in some occasions, the black drink was used in other ritual contexts—ballgames, feasting, council meetings, burial ceremonies—and it was also often used simply as an everyday social beverage. In fact, Figure 2. Distribution of black drink use among ethno-historic Native there is no evidence that there is anything Americans (Herb Roe 2008; From Merrill 1979:44) about Ilex vomitoria that actually causes vomiting. For example, it was consumed by many early colonials as a tea popularly known as cassina. It is possible that another emetic plant was added to the tea, or perhaps regurgitation occurred simply because Indians drank copious amounts of the hot fluid during these ceremonies—by some accounts over 6 gallons of the brew! Tushingham et al. 2011‐3 From the ethnographic record it seems that the black drink was used in most places where Ilex vomitoria was available (Figures 2, 3). There is evidence it was traded to interior groups, perhaps along with marine shell cups, and some groups tended patches of the plant in locations where it would not normally grow. PREHISTORIC USE Figure 3. Natural Range of Ilex vomitoria Aiton (Merrill 1979:42) Prehistoric use of Ilex vomitoria and the emergence of the black drink ritual is poorly understood. It is often associated with marine shell cups, especially those found in mortuary contexts. As shell cups or dippers were used by historic Indians as black drink containers or drinking vessels, it is thought that they may have had a similar use in the past. Also, as shell cups were often buried with individuals of a certain stature (Figure 4), their presence in prehistoric mortuary contexts is similarly linked to high status ceremonialism. Some of the best evidence for some sort of drinking ritual, possibly involving the black drink, dates to the Mississippian period. It is most famously associated with shell cups engraved with Southeastern Ceremonial Complex iconographic symbols (Figure 5). Occasionally these cups and associated pottery are found with a black residue on their interior walls. In a 1979 survey of possible archaeological evidence for the black drink Jerald Milanich hypothesized that the origins of the black drink or some sort of drinking ritual date back earlier than the Southeastern Ceremonial Complex—perhaps as early as Hopewell. He based this assertion on the fact that the cups were widely distributed in mortuary and/or ceremonial contexts similar to those observed in the ethnographic record. PLANT BIOLOGY Yaupon (Ilex vomitoria) is a dioecious shrub or small tree species of the holly family (Aquifoliaceae). This species is native to the coastal plain of the southeastern United States where it is distributed along the Atlantic coast from southern Virginia to central Florida and on the Gulf Coast from central Florida to east Texas. Determining the precise boundaries of this range is confounded by the probable Figure 4. Burial of Timucan warriors in a mound with weapons and shell cups as grave goods. From De Bry, Florida, pl. XIX. Smithsonian Institution, National Anthropological Archives, dispersal of yaupon by Native American trade to negative no. 57-559. Tushingham et al. 2011‐4 areas beyond its natural distribution (Catesby 1974, Brickell 2000, MacDougal 2003). Such evidence for human dispersal of yaupon makes it difficult to determine whether yaupon’s distribution beyond the coastal plain is of natural or human origin. Within its native range, yaupon grows in coastal scrub, pineland, and hardwood forest habitats (Johnson and Barbour 1990). Yaupon foliage is consumed by numerous taxa of insect herbivores including a specialist holly leaf‐miner (Phytomyza vomitoriae) and both leaves and twigs are browsed by white‐tailed deer (Odocoileus virginianus). In spring (March‐ April), yaupon produces small white flowers that are insect pollinated (mainly hymenopteran species) and its fruit is a bird‐dispersed drupe that turns red at maturity. Ilex vomitoria is a psychostimulant plant that contains caffeine. Although another holly species, Ilex cassine, produces small quantities of caffeine, yaupon is the only known North American plant species that produces significant quantities of caffeine and is the source of cassina, the stimulating Amerindian beverage (Edwards and Bennett 2005). Two South American congeners produce caffeine, Ilex paraguariensis and I. guayusa. It is thought vomitoria has its origins from the south. Mean foliar, dry‐weight caffeine concentrations of wild‐type yaupon