Current research on smoking pipe residue

Journal of Archaeological Science 39 (2012) 1951e1959 Contents lists available at SciVerse ScienceDirect Journal of Archaeological Science journal homepage: http://www.elsevier.com/locate/jas Current research on smoking pipe residues Sean M. Rafferty*, Igor Lednev, Kelly Virkler, Zuzana Chovanec University at Albany, Department of Anthropology, 1400 Washington Ave., Albany, NY 12222, USA a r t i c l e i n f o a b s t r a c t Article history: Received 1 February 2011 Received in revised form 30 January 2012 Accepted 2 February 2012 This paper presents research into the identification of tobacco residues in ancient smoking pipes. Two techniques have been used so far: gas chromatography/mass spectroscopy (GC/MS), and Raman microscopy. GC/MS has been used successful in the past by the author to identify ancient tobacco residues, and the results of this round of analysis support prior research. Raman microscopy, which has the advantage of working on dry samples without solvents, was not successful. It appears that combustion products overwhelm any useful signal that would identify the substance smoked. We are pursuing the use of Raman in non-combusted samples. Ó 2012 Published by Elsevier Ltd. Keywords: Residue analysis Raman microscopy GCeMS Tobacco 1. Introduction This paper presents recent research results on smoking pipe residues using two analytical techniques: Raman microscopy and gas chromatography/mass spectroscopy (GCeMS). This focus of the research represents is the application of residue analysis to the detection of intoxicant compounds. These two analytical methods are applied to both experimental preparations of smoking pipe residues as well as to a sample of archaeological residues. Research result so far indicate that, while Raman microscopy has significant long term potential for archaeology, additional methodological development is needed for its widespread application to organic residues. GCeMS continues to be a viable technique for the analysis of smoking pipes, and current results are presented. 2. Research questions Previous research on this topic used gas chromatographyemass spectroscopy (GCeMS) to identify nicotine degradation products in three archaeological pipe specimens: a nineteenth-century pipe from Ghana, and two Early Woodland Period pipes from Eastern North American sites dating to between 2300 and 2500 B.P. (Rafferty, 2002, 2006). The current stage of research investigates two research questions. The first is the potential for using Raman microscopy on smoking pipe residues. Raman microscopy is a highly accurate technique that requires minimal preparation of samples. Raman * Corresponding author. Tel.: þ1 518 442 4713. E-mail address: [email protected] (S.M. Rafferty). 0305-4403/$ e see front matter Ó 2012 Published by Elsevier Ltd. doi:10.1016/j.jas.2012.02.001 can detect the presence of compounds of interest directly from the solid phase, in situ on an artifact’s surface. Growing interest in application of Raman spectroscopy in art and archaeology during the last decade is well described in a recent review (Vandenabeele et al., 2007). Current developments of Raman spectroscopy at the University at Albany department of chemistry (Lednev et al., 2005, 2006) have further improved the technique by allowing the analysis of heterogeneous samples through statistical analysis (Shashilov et al., 2006) of multiple observations of a single sample. This is ideal for archaeological samples, which are rarely homogenous. Raman microscopy has been applied in recent years to a number of archaeological case studies, rock art and tomb paintings (Smith and Barbet, 1999), ceramic pastes and glazes (Bruno et al., 1997; Clark and Curri, 1998), ancient glass (Bouchard and Smith, 2002), stone tools and ancient metals (Gendron et al., 2002; McCann et al., 1999; Smith and Gendron, 1997), ancient textiles and pigments (Edwards et al., 1997; Wiedemann et al., 2002), and ancient organic residues (Edwards et al., 1998), including residues associated with opium (Schultz et al., 2004). Results of the research presented herein indicate that, while Raman microscopy is a powerful technique, there are some unique challenges posed in analyzing organic residues. The second research question was does additional research support previous research on smoking pipes. Previous GCeMS research (Rafferty, 2002, 2006) indicated an early adoption of tobacco in the Eastern Woodlands. That assessment was based on a small sample, and it was important to see if additional specimens would corroborate earlier results. Results of this research show corroboration for one pipe sample, but more equivocal results for two others. 1952 S.M. Rafferty et al. / Journal of Archaeological Science 39 (2012) 1951e1959 3. Current research Initial use of Raman microscopy was promising. Standards of pure nicotine were prepared on foil targets. Three samples were prepared; two were subjected to artificial aging by heating and ultraviolet light exposure. Clear spectra were recovered for the unmodified and aged samples, with slight differences between them (Fig. 1). This showed that Raman can identify nicotine, and that it may be useable to determine between “fresh” and aged samples of nicotine. The next step was to prepare samples of tobacco residue through controlled combustion of samples of Nicotiana rustica “Native Tobacco” and analyze the residue via Raman to ascertain if nicotine was detectable. It was here that we ran into serious problems. At first it seemed that we had a clear spectrum for tobacco residue. However, we checked this spectrum against one derived from burned paper and found the two spectra to be identical. This indicated that either general combustion products of organic material, or the fluorescence of the residue, or both, overwhelm any nicotine present in the sample. In hindsight this is not surprising as nicotine is present in minute abundance even in fresh tobacco leaf. I will note in brief that the opium aspect of the research also met similar problems using Raman. These negative results raise concern about the suitability for Raman for the analysis of organic residues. Consultation with the co-Principal Investigator for the NSF research concluded that fluorescence is a serious problem for darker colored substances. Given that most organic residues are by nature dark in color, this could prove a serious liability for the technique. As we were at an impasse with regards to Raman microscopy, we turned to “conventional” residue analysis techniques using GCeMS, which had proven successful in the analysis of North American artifacts in recent analyses (Reber et al., 2010; Tushingham et al., 2010). This research took the same approach as the Raman analysis, in that the technique was first applied to experimentally produced residues to verify efficacy, and then to archaeological contexts. GCeMS analysis followed a modified version of previously successful protocols (Rafferty, 2002, 2006), themselves adapted from Gager (Gager, 1991) and by Zahlsen and Nilsen (Zahlsen and Nilsen, 1994). Alkaloids were extracted from residues through the Fig. 1. Raman spectrum of nicotine. elution of methylene chloride. The resultant solution was then concentrated via evaporation under nitrogen flow. The concentrated solution was then injected into the GC/MS instrument (HP 6890 GC, 5972A Mass Selective Detector, 1 ml auto-injector, HP-5MS 30m  0.25mm  250 mm column). GC settings started at 150  C for 2 min, then ramped up at 10  C/mine180  C, for a total run time of 5 min. Due to the small abundances of the compound being analyzed, MS proceeded using select ion monitoring for four ions (m/z of 84, 133, 161 and 162 respectively). While this is somewhat different equipment than had been used in past analyses (most significantly an HP-5 column as opposed to an HP-1 column), it is shown below that the instrumentation was effective in detecting nicotine in organic residues of varying ages. First, standard solutions of pure nicotine were analyzed; in all cases the instrumentation was able to identify nicotine at a retention time of 2.7 min (Fig. 2). This shows that the instrumental set up is effective at detecting nicotine in organic residues. It should be noted that this is a significantly shorter retention time than has been identified in previous research. This was not intentional on the part of the researchers, as a measure to allow more rapid analysis for example. Rather, it was an unavoidable side effect of using different instrumentation. Regrettably we were limited to instrumentation available in the University at Albany Department of Chemistry. Second, artificially prepared N. rustica residues were analyzed. Residues were prepared by reverse airflow combustion in a labproduced ceramic pipe with paste and temper characteristics congruent with prehistoric Native American examples (i.e., air was blown through the bowl of the pipe, with the smoke being directed into a fume hood). This created a viable pipe residue without the necessity of smoking the tobacco directly or exposing researchers to “second hand” smoke. Nicotine was again identified in this specimen (Fig. 3). These initial steps ensured that the extraction and detection protocols were appropriate for the detection of nicotine from burnt organic residues. Once the extraction and detection protocols were known to be functioning as expected, the technique was applied to archaeological samples. The first of these was a mid-18th century specimen from New Jersey. Nicotine was again detected in this archaeological sample (Fig. 3). Given the relatively recent date of the specimen and the clear Euroamerican manufacture, it was expected that this pipe was used for tobacco, and the successful detection of nicotine shows that tobacco residue remains detectable in archaeological contexts over relatively short periods of time. Finally, three pipe specimens of considerably greater age were analyzed. These pipes were recovered from the Mathies Mound site, the Quaker State Rockshelter, and the Glenshaw Rockshelter, all located in Western Pennsylvania (Fig. 4). Residue samples from these pipes were generously donated by the Carnegie Museum of Natural History where all three specimens are curated. All three are stylistically similar to specimens dated to the Early Woodland Period, from approximately 2,000 to 3000 B.P. (Fig. 5). A radiocarbon date of 2541  51 B.P. was derived for the Quaker State Rockshelter pipe (AA89866). This date yields a calibrated two sigma range of between 809 and 510 B.C.; the Adena affiliation of the site would indicate an origin towards the recent end of that range, most likely during the 6th century B.C. There was not enough residue for a date for Mathies Mound, and regrettably a radiocarbon sample taken for the Glenshaw Rockshelter was lost in transit to the lab. All three of these residue samples tested positive for the presence of nicotine, though the quality of the results varies among the samples. The Mathies Mine site had the clearest chromatographic peak (Fig. 6). While the abundance for this peak is relatively low, presumably due to the extreme age and small size of the sample, S.M. Rafferty et al. / Journal of Archaeological Science 39 (2012) 1951e1959 1953 Fig. 2. GCeMS spectrum of nicotine. the peak is discrete and is located at the right retention time for nicotine. The Glenshaw Rockshelter and Quaker State Rockshelter had less clear chromatographic peaks, with the nicotine peak blending into a “plateau” on the chromatogram (Figs. 7 and 8). This feature does begin at 2.7 min, the retention time for nicotine, and does produce a positive mass spectrum identification for nicotine as well. While it is plausible that tobacco was smoked in these two pipes, the data is supportive, but not conclusive. A caveat must be included at this point that the chromatographic peak that showed a mass spectrum of nicotine was of 1954 S.M. Rafferty et al. / Journal of Archaeological Science 39 (2012) 1951e1959 Fig. 3. GCeMS spectrum for an 18th century smoking pipe. extremely low abundance in all three cases, and that other, nonnicotine compounds that also include the four target ions could give false positives. However, the GC peak is at the same retention time (2.7 min) for nicotine derived from our control samples of pure nicotine, and the MS identifications were at confidence levels of over 80%; false positives are generally of much lower confidence for this instrument. We are therefore reasonably confident that tobacco was smoked in each of these pipes. Regrettably the available samples were all very small, less than 0.1 g each, and all were totally expended in the residue analysis and AMS dating described in this article. 4. Discussion Raman microscopy is a powerful analytical technique, with great potential for archaeological analysis. However, based on our current results the technique is not yet able to detect lowabundance compounds in heterogeneous organic residues. We are currently pursuing research on other substances, such as honey or blood, where we expect a lower chance of sample fluorescence. Our research shows that current GCeMS instrumentation and existing analytical protocols are appropriate for the detection of nicotine, and by extension, of tobacco, in prehistoric residues. This was demonstrated by the detection of nicotine experimental standards and artificial residues, as well as a relatively recent archaeological sample. When applied to a sample of much older archaeological samples, the technique provided results which support previous research indicating an Early Woodland introduction of tobacco into the Eastern Woodlands, between 2500 and 3000 B.P. This is significant as the current state of ethnobotanical data for tobacco dates to the first century A.D., centuries after the dates of the smoking pipes analyzed in this research program so far. The earliest archaeobotanical evidence of the use of tobacco in eastern North America comes from the central Mississippi Valley between A.D. 100 and 200 (uncalibrated) (Asch, 1991; Asch, 1994; Haberman, 1984; Wagner, 2000; Winter, 2000a), with dates for the rest of Eastern North America falling several centuries later (Haberman, 1984; Von Gernet, 1992). Recently derived dates from the Southwest are earlier, with specimens from New Mexico dated to 1040 B.C. (Bohrer, 2004; Winter, 2004). This indicates that tobacco was introduced into the Southwest through Mexico along the same route as cultigens such as maize, and reached the Eastern Woodlands through the Great Plains, most likely sometime in the mid-first millennium B.C. based on our residue analysis data. Documenting the early origin and spread of tobacco use is more than a botanical curiosity. N. rustica was used by Native Americans in eastern North America at the time of contact, and has the highest nicotine content of any tobacco species. Nicotine causes tachycardia, vasoconstriction, and increased alertness. In large doses nicotine has hallucinogenic effects (Joniger and Dobkin de Rios, S.M. Rafferty et al. / Journal of Archaeological Science 39 (2012) 1951e1959 1955 Fig. 4. Location of prehistoric smoking pipes analyzed. 1973; Joniger and Dobkin de Rios, 1976). Tobacco is safer than other Eastern North American psychoactive plants such as Datura, and may have been adopted as a foreign alternative to potentially dangerous local intoxicants (Goodman, 1993; Von Gernet, 1992; Winter, 2000b). Tobacco’s allure to prehistoric peoples was likely exacerbated by the lack of any significant indigenous history of fermentation of alcoholic beverages, which is the most common type of intoxicant across most of world cultures today. While there has been significant research on the prehistory of tobacco use in North America, there are still questions regarding the chronology and geographic patterning of tobacco’s adoption. Ethnohistoric evidence indicates that the use of tobacco was Fig. 5. Images of smoking pipes analyzed. 1956 S.M. Rafferty et al. / Journal of Archaeological Science 39 (2012) 1951e1959 Fig. 6. GCeMS spectrum of Mathies Mine pipe. ubiquitous to Native American cultures at the time of contact (Springer, 1981). A practice with such a wide geographic distribution should have deep prehistoric roots, but the chronology of its origin and spread is still poorly understood due to a lack of botanical data. The effects of a particular intoxicant compound have a major effect on the sociocultural aspects of its use (Dobkin de Rios, 1975; Dobkin de Rios, 1984). The effects of high doses of nicotine, such as “out of body” or flying sensations, presumably affected ritual practices in prehistoric Eastern North America, especially bird representations exhibited found I a range of artistic media (Dobkin de Rios, 1976). Our research into the prehistory of tobacco use through residue analysis evidence shows that the early history of tobacco may be longer than previously supposed. Some researchers have proposed that tobacco was recently introduced into Eastern North America (Ford, 1981; Knight, 1975; Watson, 1989; Yarnell, 1964). Were this the case, it would imply that smoking pipes predating predate the second century A.D. would have been used to smoke something other than tobacco. The results of our research show that such assumptions need further evaluation. Limitations in the research program remain, and will provide directions for future investigations. We have not yet tackled the problem of correlating abundance of nicotine in experimental or archaeological residues, with the concentration of nicotine in the original plant smoked. While it may be possible to make this correlation for experimental residues, where the starting concentration of nicotine in the tobacco plant tissue can be S.M. Rafferty et al. / Journal of Archaeological Science 39 (2012) 1951e1959 1957 Fig. 7. GCeMS spectrum of Quaker State Rockshelter pipe. known in advance, the lack of this starting point datum, as well as a wide variety of taphonomic processes that would degrade the abundance of nicotine over time, may make it very difficult if not impossible to determine this relationship for archaeological samples. This is significant, in that it will likely limit our ability to determine empirically the actual species smoked in prehistoric pipes based on residue analysis. While ethnobotanical evidence indicates N. rustica was used in the Eastern Woodlands in the early First Millennium A.D., we will likely have to wait for additional botanical samples to extend this date earlier to meet the date for a general species of tobacco based on residue analysis. In short, we know prehistoric Native Americans were smoking tobacco, we just can’t yet confirm that the species was N. rustica. We have also not determined what plants other than tobacco, if any, may have been smoked in prehistoric Native American 1958 S.M. Rafferty et al. / Journal of Archaeological Science 39 (2012) 1951e1959 Fig. 8. GCeMS spectrum of Glenshaw Rockshelter pipe. pipes. Ethnohistoric accounts indicate a variety of plants that were smoked in addition to tobacco, including Cornus sp. (Dogwood), Juniperus species (Juniper), Rhus glabra (Sumac) and Arctostaphylus uva-ursi (Bearberry) (Brown, 1989; Hall, 1977; Springer, 1981; Yarnell, 1964). The limitation is that the use of select ion monitoring for GCeMS requires the researcher to look for the presence/absence of compounds, rather than scan for all compounds. Active compounds must be determined in advance, and then compared to compounds in a sample, to make comprehensive determinations of all plants smoked. Research to identify biomarkers of the above listed tobacco alternatives is currently ongoing. 5. Conclusions Residue analysis of intoxicants such as alkaloids is a profitable, if under-investigated, topic of research (Wink, 1998). There have been recent examples of intoxicant research, on substances as varied as S.M. Rafferty et al. / Journal of Archaeological Science 39 (2012) 1951e1959 cacao (Hurst et al., 2002, 1989), opium (Bisset et al., 1996; Koschel, 1996), as well as other researchers looking at tobacco (Tushingham et al., 2010). Intoxicants play roles in a variety of significant cultural functions, such as religious rituals or social feasting (Bourguignon, 1973; Merlin, 2003; Rudgley, 1993; Winter, 2000b). Residue analysis can provide data on such practices (Rafferty, 2007). Additional research will seek to further corroborate and add detail to these results. Acknowledgements This research was funded in part by the National Science Foundation (BCS-0822493). Radiocarbon assays were performed at the University of Arizona AMS Radiocarbon lab. Special thanks to William Tippens and the curatorial staff of the Carnegie Museum of Natural History for access to collections and photographs of specimens. Thanks to Colin Henck of the University at Albany’s department of chemistry for assistance in modifying the GCeMS protocols. References Asch, D.L., 1991. Tobacco Seeds in the Archaeobotanical Collections of the Center for American Archaeology. Center for American Archaeology, Kampsville. Asch, D.L., 1994. 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