Journal of Archaeological Science 32 (2005) 957e965 http://www.elsevier.

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Fatty acid analysis of prehistoric burned rocks: a case study from central California
Tammy Buonasera*
Archaeological Research Program, California State University, Chico, 25 Main Street, Suite 101, Chico, CA 95929-401, USA Received 19 November 2004; received in revised form 14 January 2005

Abstract Although lipid analysis of archaeological residues has been utilized for almost three decades, it has rarely been applied to archaeological materials other than pottery. A 2001 study used lipid analysis of burned rocks (sandstone) and ground stone tools to interpret subsistence change at a south Texas site (M.J. Quigg, M.E. Malainey, R. Przybylski, G. Monks, No bones about it: using lipid analysis of burned rock and groundstone residues to examine Late Archaic subsistence practices in South Texas, Plains Anthropologist 46 (2001) 283e303). If such applications are reliable, insight into past subsistence practices could be greatly enhanced. The following study tested whether measurable amounts of lipids could be extracted from burned rocks (andesite) from a central California site and the ability of fatty acid analysis to reliably interpret those extracts. While results indicate that some of the burned rocks may contain lipids absorbed from cooking activities, lipids were also recovered from o-site rocks. Before reliable interpretations of culturally introduced lipids can be made, more thorough study of the types and amounts of lipids present in rocks due to natural processes is necessary. 2005 Elsevier Ltd. All rights reserved.
Keywords: Gas chromatography; GC; Lipids; Fatty acids; Burned rocks; Fire aected rocks

1. Introduction Archaeology integrates a wide variety of information in its eorts to reconstruct and analyze past human behavior. Increasingly, biomolecules such as nucleic acids, proteins, and lipids are providing new sources of data for the study of past cultures [8,25,26,29,32]. Since 1976 [1], lipid analysis via gas chromatography (GC) [19e22,27,28] and gas chromatography-mass spectrometry [2,4e7] have been used to identify various plant and animal residues absorbed in pottery sherds. Although lipid analysis has typically focused on pottery, the archaeology of regions and time periods lacking pottery may also be able to benet from its application. Nonceramic materials that may be amenable to lipid analysis
* Tel./fax: C1 530 898 5554. E-mail address: tbuonasera@mail.csuchico.edu 0305-4403/$ - see front matter 2005 Elsevier Ltd. All rights reserved. doi:10.1016/j.jas.2005.01.012

include milling tools, stone bowls or other types of stone vessels, and cooking rocks. A 2001 study by Quigg et al. [28] used GC to analyze fatty acids (a type of lipid) from burned rocks and ground stone tools recovered from a Late Archaic Period site in south Texas. The study identied various plant and animal sources using a set of experimentally determined criteria based on the relative percentages of 10 fatty acids. Results were promising. Almost all rocks yielded measurable amounts of lipids, and provided subsistence information for a site lacking faunal or macrobotanical remains. However, the Quigg et al. [28] study made no mention of testing for amounts or types of lipids that may have been present in non-culturally associated rocks, and seemed to assume that any lipids recovered were the result of human use (i.e. cooking). Unlike the situation with ceramics, lipid analysis of stone materials has not

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been widely tested. It is not known what capacity various rocks have for absorbing lipids, or what quantities and types of lipids may be present in rocks due to natural processes (i.e. absorption from surrounding soil and decaying organic material). In order to accurately interpret results, it may be important to know what types and quantities of lipids are naturally present in rocks. The following study was performed as an initial step in exploring the potential of lipid analysis to provide subsistence related data from burned rocks. The rst question addressed was whether analyzable quantities of lipids could be extracted from re aected andesitic rocks recovered from a Late Prehistoric rockshelter in central California. The second question addressed was whether any such lipids may have been introduced through various cooking activities. These re aected rocks had previously been interpreted as cooking rocks and may have served a variety of functions involving hearths [33]. Prehistoric populations in central California did not use ceramic vessels for cooking. Early ethnographers recorded the use of a stone boiling technique for preparing acorn mush or soup [17,24]. In this method, heated rocks were added to a cooking basket containing water and leached acorn meal. Rocks were removed from the basket as they cooled, and new ones were added, until the mixture reached the desired degree of cooking. Pine-nut soup was prepared in a similar manner [9]. Heated rocks were also used to construct earthen ovens for baking acorn bread and a variety of other foods [17]. If the FMRS burned rocks were used in these types of cooking activities, lipids from foods they were in direct contact with might be absorbed and preserved within the rock matrix by mechanisms similar to those proposed for ceramics [8,31]. Because rocks extracted in the Quigg et al. [28] study were sandstone (presumably more porous than andesite) it was important to rst determine whether measurable quantities of lipids could be extracted from the andesitic rocks. Next, in order to assess the likelihood that any lipids present in the burned rocks were the result of cooking activities, as opposed to absorption from the depositional environment, seven cooking rocks and three o-site rocks of similar type were extracted and analyzed using GC. Fatty acid compositions (relative percentages) as well as total fatty acid concentrations (total standardized area counts) were compared.

2. Lipids, fatty acid analysis, and GC Lipids are a broad class of molecules dened by their solubility. Lipids are insoluble in water but soluble in organic solvents; they include fats, waxes, oils, terpenes,

and steroids [23]. The lipids this study focused on are fatty acids. Fatty acids are long chain carboxylic acids that are an essential component of many lipids [11,15]. Structurally, fatty acids are often divided into saturated and unsaturated forms. The carbon chains in saturated fatty acids are fully substituted with hydrogen, while unsaturated fatty acids have one or more carbone carbon double bonds. The shorthand designation for fatty acids shows the number of carbons, followed by the number of carbone carbon double bonds. For example, oleic acid, a fatty acid 18 carbons long with one double bond, is designated as C18:1. This notation can be expanded to indicate positions of double bonds [11,15]. Lipids are useful biomarkers for archaeology because they possess a greater potential for preservation than most other classes of biomolecules [8,14]. In addition, many organic materials have characteristic fatty acid proles or other diagnostic lipid components [5,7,8,12]. Thus, fatty acid analysis is used in food science to determine food purity, as well as in biology to address taxonomic issues [3,12]. Unfortunately, direct comparisons between the relative percentages of fatty acids in modern foods and ancient residues cannot be made. This is because degradation alters the prole of fatty acids in a manner that changes their relative amounts. Certain fatty acids, typically unsaturated fatty acids, are more susceptible to processes of degradation than others [8,11,31]. Over time, this will tend to increase the proportion of saturated to unsaturated fatty acids. Several studies have been conducted on the degradation of fatty acids by simulating long-term decomposition as well as eects of cooking [4,19,21,27,28,30]. The proles of both modern sources, and of experimentally aged and thermally altered residues have been assessed in order to develop various identication methods [4,19,21,27,28]. In addition to interpretive complications arising from degradation, residues extracted from archaeological materials may also have more than one source. The issue of potential food mixtures is sometimes addressed by empirically determining fatty acid proles of likely food combinations [19,20,28]. Gas chromatography is useful for analyzing archaeological residues because it can separate and detect very small quantities of material [8,20]. A mixture is injected into a heated column and molecules are separated based on their partitioning between a stationary phase (the column) and a mobile phase (the carrier gas) [11,15]. Identications are made by comparing retention times to one or more external standards. This study used GC because it is the primary method used in lipid analyses, such as the Quigg et al. study [28], that rely solely on relative amounts of fatty acids to make source identications [19e22,27]. An alternative

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and more powerful way of identifying a wider array compounds is to combine GC with mass spectrometry [4e8,31].

4. Laboratory methods Rock samples were powdered using a clean, diamond embedded steel disc tted to an angle grinder. To prevent contamination with modern lipids, the rst 1e2 mm of the rock exterior was removed prior to collecting samples. Sample sizes, descriptions, and provenience are given in Table 1. Samples were extracted using a modied Bligh and Dyer method. Although most archaeological applications of lipid analyses have used a Folch extraction method (but see Patrick et al. [27]), a modied Bligh and Dyer method has been very eective for extracting soil lipids in the laboratory where this study was performed. For a more thorough discussion of these methods see Kates [15]. Powdered rock samples were extracted by rocking (2 h) in 60 ml chloroform, methanol and citrate buer (CMC), pH 4.0 (1:2:0.8 v/v/v). The intact surfaces of two additional cooking rocks were also extracted by submerging the rocks in 100 ml CMC and sonicating, 10 min (2!). Phases were broken with one volume each of chloroform and water. The chloroform phase was collected and evaporated under a stream of nitrogen. Lipids were ushed with nitrogen and stored at 20  C until the production of fatty acid methyl esters (FAMES). Production of FAMES was accomplished by adding 3 ml of 4% sulfuric acid in methanol to the dry lipids and heating (85e90  C, 60 min). FAMES were neutralized, then extracted with 2 ml hexane (2!), dried under nitrogen and re-dissolved in 50ml chloroform. Samples were ushed with nitrogen, and stored at 20  C. To minimize contamination with modern fatty acids, all glassware was cleaned with a laboratory grade cleaner prior to use, and gloves were worn when handling samples. Samples (1 ml) were injected with a split injection ratio of 1:5 on a Varian 3800 GC with a HP-5 column. Helium was the carrier gas at a ow rate of 0.8 ml/min.

3. Site information and selection of samples Material for the current analysis came from the Fort Mountain Rockshelter (FMRS) (CA-CAL-91). This rockshelter is located at 3300 feet elevation on the western slope of the Sierra Nevada, and was excavated in 1987 [33]. The FMRS contains three temporal components. The oldest component is only present on the debris apron and was formed pre AD 1500; this component is thought to represent one or more logistically organized hunting episodes [33]. Midden from the two younger components is located on the shelf as well as the upper 20 cm of the debris apron. Many charred pine cone scales identied as Gray Pine (Pinus sabiniana), charred acorn hulls, and acorns (species unknown) were recovered from the shelf. The shelf deposits represent longer episodes of occupation, and have been dated to 250G60 years BP [33]. Several factors made materials recovered from this site desirable for fatty acid analysis. First, it is a dry, sheltered deposit, facilitating organic preservation. Second, a cache of unburned acorns was discovered on the shelf, providing an opportunity to see how the fatty acid prole of acorns might change over time (results from this comparison will be reported in a future study). Third, numerous re aected rocks, thought to be associated with cooking activities, were recovered from the site. Rock samples for the following analysis were selected from both the debris apron and the shelf midden. Osite rocks, similar in size and shape to the FMRS samples, were collected from a forested location near Strawberry Valley, California. Like the FMRS samples, these were andesitic rocks from the Mehrten formation.

Table 1 Sampling data Sample FMRS 3 FMRS 6 FMRS 7 FMRS 8 FMRS11 FMRS12 FMRS24 FMRS25 FMRS26 Rock 1 Rock 2 Rock 3 Provenience S0.3/E4.5 (0e10 cm) shelf S5/W0.75 (0e10 cm) apron S5/W0.75 (10e20 cm) apron S5/W0.75 (20e30 cm) apron S3/W3.5 (0ebdr) shelf S1.5/E2.5 (0e10 cm) shelf N1/E4.5 (10 cmebdr) shelf S0.3/E4.5 (0e10 cm) shelf E2/N0 (20e30 cm) shelf Mehrten formation, Strawberry Valley, CA Mehrten formation, Strawberry Valley, CA Mehrten formation, Strawberry Valley, CA Rock dimensions (cm) 6.0!4.5!5.0 Not recorded Not recorded 8.0!3.8!3.5 Not recorded Not recorded 7.5!5.8!3.4 8.0!7.0!5.5 6.0!8.5!3.8 6.0!5.8!3.4 7.0!6.6!6.4 11.5!8.0!3.4 Sample wt. (g) e 37.6 40.3 e 41.4 46.9 29.5 26.0 21.1 53.7 47.1 42.3 Extraction method Surface Powdered Powdered Surface Powdered Powdered Powdered Powdered Powdered Powdered Powdered Powdered interior interior interior interior interior interior interior interior interior interior

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T. Buonasera / Journal of Archaeological Science 32 (2005) 957e965 Table 2a Criteria for the identication of archaeological residues based on the decomposition patterns of experimental cooking residues prepared in pottery vessels (from Quigg et al. [28]) Identication Large herbivore Large herbivore with plant or bone marrow Plant with large herbivore Beaver Fish or corn Fish or corn with plant Plant (except corn)
a

Temperature was programmed from 80e140  C at 20  C/min, 140e224  C at 4  C/min, held for 5 min at 224  C, and ramped to a nal temperature of 265  C at 4  C/min, while the injection port was maintained at 250  C. With the exception the two rock surfaces, all samples were analyzed on the same column, under the same parameters. Peaks were identied by comparison to appropriate external standards (Supelco). An internal standard was not used, as previous studies using GC only [19e22,27,28] have relied on peak areas to calculate relative amounts of fatty acids, and have not reported absolute amounts of fatty acids. 5. Analysis Cooking rocks from the FMRS were compared with o-site rocks. Dierences among the seven powdered FMRS samples and the three powdered o-site samples were assessed by comparing the total concentration, as well as the relative percentages of 10 fatty acids (C12:0, C14:1, C14:0, C15:0, C16:1, C16:0, C17:0, C18:2, C18:1 isomers, and C18:0). These are the same fatty acids that were used by the Quigg et al. study [28] to determine the source(s) of lipid residues extracted from archaeological materials. Results were compared by the ManneWhitney U test. Because sample size was small and normality could not be assumed, the ManneWhitney U-test was used as a non-parametric substitute for the independentsample t test. According to Zar [34], the ManneWhitney is about 95% as powerful as the independent-sample t test. Dierences were considered signicant at p!0.05. Statistical analyses were performed using SPSS 12.0. Two dierent sets of criteria were applied to identify sources of fatty acids extracted from samples. One method, used in the Quigg et al. study [28], is based on a set of criteria developed by Malainey [18e20]. These criteria were determined for plant and animal taxa important to prehistoric populations in western Canada, and were based on the percent composition of 10 fatty acids recovered from experimental sherds that had been exposed to heating and simulated aging. The other method is a percent saturation value (%S ) determined by Marchbanks [21]. This is an earlier and more generalized method for identifying broad resource categories (i.e. plant, sh, land animal). Marchbanks %S was determined for modern residue samples, with the expectation that %S values will tend to increase with age. Identication criteria for both methods are listed in Tables 2a and 2b. 6. Results Relative percentages of fatty acids, along with the standardized total peak area (total peak area/sample

C12:0CC14:0 CC15:0 15% Lowa

C18:0 R27.5% R25%

C18:1 15% 15%x25%

R15% Low Low R15% R10%

R25% Low 25% 25% 27.5%

No data R25% 15%x27.5% 15%x27.5% 15%

Low was interpreted as lower than the highest value giving a positive identication in Quigg et al. [28].

weight), are shown in Table 3. All samples (FMRS rocks and o-site rocks) yielded measurable amounts of fatty acids. Reagent blanks contained very low levels of fatty acids. The two intact surface extractions (FMRS 3, and FMRS 8) were analyzed on a dierent column than all the other samples, and had much lower total area counts than the powdered samples. Because these factors were likely to have aected the relative percentages of the ten fatty acids, FMRS 3 and FMRS 8 were not included in subsequent analysis. When compared as two groups, the FMRS samples were found to not dier signicantly from the o-site samples in total standardized fatty acid concentration ( p-values are reported in Table 4). However, three of the FMRS samples (7, 12, and 26) had lipid concentrations two to six times greater than that of the highest o-site rock sample (Table 3 and Fig. 1). Dierences in lipid composition (relative percentages) were signicant for three of the 10 fatty acids; the FMRS samples had a signicantly higher proportion of both C15:0, and C16:1, and a signicantly lower proportion of C18:2 than the o-site rocks (Table 4 and Fig. 2). The gas chromatogram of FMRS7 is shown in Fig. 3. Identications made using Malaineys criteria and Marchbanks %S are provided in Table 5. One of the FMRS samples and one of the o-site samples fell outside the range of identiable fatty acid combinations specied by the Malainey criteria. Of the remaining six
Table 2b Percent saturation (%S=C12:0CC14:0/C12:0CC14:0CC18:2CC18:3) for modern plant and animal sources (adapted from Marchbanks [21]) Source Plants Fish Land animals %S 0e18 22e40 47e98

T. Buonasera / Journal of Archaeological Science 32 (2005) 957e965 1.15 0.30 4.55 1.03 0.36 21.32 0.98 14.25 34.20 21.87 6669 Rock 3 Table 4 Mean ranks and p-values for FMRS samples vs. o-site rocks Group Area counts per g C12:0 C14:1 0.89 0.26 3.85 1.17 0.30 32.61 0.87 11.05 34.16 14.93 4134 Rock 1 C14:0 C15:0 C16:1 4.75 0 0 1.95 26.20 2.17 0 0 58.20 6.74 e C16:0 C17:0 FMRS 3a C18:2 0.03 0.23 7.69 3.83 3.55 4.63 0 20.74 21.41 37.89 e C18:1 C18:0 FMRS 26 Table 3 Relative percentages of 10 fatty acids and standardized total area counts for FMRS samples and o-site rock samples 3.55 0.12 5.33 1.50 0.58 22.96 4.06 6.44 37.38 18.07 42293 Off-site rocks FMRS samples Off-site rocks FMRS samples Off-site rocks FMRS samples Off-site rocks FMRS samples Off-site rocks FMRS samples Off-site rocks FMRS samples Off-site rocks FMRS samples Off-site rocks FMRS samples Off-site rocks FMRS samples Off-site rocks FMRS samples Off-site rocks FMRS samples N 3 7 3 7 3 7 3 7 3 7 3 7 3 7 3 7 3 7 3 7 3 7 Mean rank 3.33 6.43 4.67 5.86 3.33 6.43 5.00 5.71 2.33 6.86 2.33 6.86 4.00 6.14 5.67 5.43 9.00 4.00 5.67 5.43 4.67 5.86

961

p-value 0.183 0.667 0.183 0.833 0.033 0.033 0.383 1.000 0.017 1.000 0.667

FMRS 8a

0.51 0.15 1.50 0.29 0.87 13.38 0.23 17.37 58.38 7.30 4710

Rock 2

FMRS 12

FMRS 11

1.79 0.29 4.03 2.64 4.28 28.01 0.44 3.18 38.13 17.22 26984

FMRS samples, ve were identied as beaver and one as sh or corn. Two o-site rocks were identied as beaver by the Malainey criteria. Because Malaineys criteria were determined for resources important to other regions, they are not representative of some important pre-contact food sources for central California. For example, they do not include acorn, but do include corn. Yet, it was anticipated that categories such as plant or large herbivore would be inclusive enough to be applicable. Marchbanks %S identied three of the FMRS samples as plant, three as sh, and one as land animal. Of the three o-site rocks, two were identied as plant and one as sh.

FMRS 25 FMRS 24

0.54 0.37 0.36 1.26 1.24 22.64 0.35 9.37 45.99 17.87 3855

2.67 0.92 9.54 2.22 1.73 15.40 0.34 6.76 51.16 9.27 6521

3.05 0.53 5.81 3.41 1.31 35.32 1.06 3.72 28.78 17.01 10630

7. Discussion In answer to the rst question posed by this study, it was found that analyzable quantities of lipids could be extracted from andesitic rocks. Furthermore, 20 to 50 g of powdered rock seems to be a greater sample size than necessary. Because samples were analyzed on the GC with a split ratio of 1:5, it seems reasonable to expect that 5 g samples would provide adequate lipids for future analyses. The second question addressed was whether the lipids present in the burned rocks may have been introduced through cooking activities. If so, it was expected that the burned FMRS rocks would contain signicantly greater amounts of lipids and dierent fatty acid compositions than the o-site rocks. When compared as two groups, the FMRS rocks were not found to be signicantly

FMRS 7

0.35 0.48 2.89 1.07 2.58 27.89 0.10 5.58 39.29 19.79 14315 C12:0 C14:1 C14:0 C15:0 C16:1 C16:0 C17:0 C18:2 C18:1 C18:0 Total area/ sample wt.
a

Fatty acid

Intact surface extractions.

FMRS 6

0.23 1.35 0.09 3.65 5.93 37.10 0.27 4.79 27.04 19.55 3849

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Fig. 1. Fatty acid quantities in FMRS samples and o-site rock samples.

dierent from the o-site rocks in total fatty acid concentrations (Table 4). However, the FMRS samples had a high degree of variability, and it is interesting to note that three of the FMRS rocks (7, 12, and 26) had quantities of lipids more than two to six times greater than the o-site samples (Table 3 and Fig. 1). It could be the case that only a few (or none) of the selected burned rocks were used in a way that placed them in direct contact with an abundant source of lipids. Between the two groups, three of the 10 fatty acids were found to have signicantly dierent relative percentages (Table 4). Relative percentages of C15:0 and C16:1 were higher in the FMRS samples than the o-site samples. This could be due to dierences in soil

Fig. 2. Relative percentages of three fatty acids in FMRS samples and o-site rock samples.

microora at the two sampling locations, because both C15:0 and C16:1 are common bacterial fatty acids [11,15] (although C15:0 is also a component of ruminant lipids [4] and C16:1 is found in plants [11,15]). The polyunsaturated fatty acid C18:2 (found in high concentrations among many plants [11,15]) was present in a lower proportion in the FMRS rocks than in the osite rocks. Again, this may be due to dierences in the biotic communities at the two sample locations. This would not be surprising, given that the FMRS rocks were recovered from a rockshelter, with very little in the way of green plants, and the o-site rocks were recovered from a more open location where there was an over-story of conifers and a sparse under-story of grasses. Unfortunately, soil samples for the FMRS, which could have provided insight into these dierences, could not be located and were presumably discarded. Most signicantly, the present study illustrates that rocks may contain naturally absorbed lipids in combinations that fall within pre-determined identication criteria (Table 5). The Quigg et al. [28] study which analyzed cooking rocks from a Late Archaic site, made no mention of testing for lipids that might be present in the rocks due to natural processes. This could be problematic because the o-site rocks analyzed in the present study contained measurable quantities of fatty acids, and two of them were identied as beaver using the same identication criteria that were used in the Quigg et al. study (Table 5). Likewise, Marchbanks %S identied residues from the o-site rocks as either plant, or sh (Table 5). The fatty acids used to make source identications in both of these methods are found ubiquitously in nature, what makes them diagnostic is their relative distribution. However, this distribution can be altered not only by degradation and mixing of food types, but potentially by mixing with lipids absorbed from the depositional environment. One way to strengthen source assignments would be to use GC/MS to identify a wider range of lipids [4e8,31], or other organic molecules [10], that may have more limited natural distributions. In addition, to be more certain that fatty acids recovered from burned rocks are the result of cooking activities, it would be benecial to compare samples to a pre-determined base-line for natural lipid concentration. If it could be demonstrated that burned rocks, or other culturally associated rock materials (e.g. milling tools, stone vessels) contained lipids in much higher quantities than non-culturally associated rocks, source determinations could be assigned with greater condence. However, to reliably determine such a base-line would require a much more exhaustive study than has yet been done. Two prior studies have attempted to assess levels of naturally introduced lipids in rock [21,22]. Unfortunately, both studies, like the present one, analyzed only three rocks and peak areas, not

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mVolts

50
16:0

18:1s

40

18:0

30

20
18:2

10
12:0

14:0

18:3 16:1

0 15 20 25 30

Minutes

Fig. 3. Partial gas chromatogram of FMRS 7.

absolute quantities, were reported. Because peak areas can vary from one column run to the next (especially when dierent columns, equipment and parameters are used) they are not comparable between dierent analyses. In pottery, the presence of lipids prior to vessel use (i.e. from the organic material present in the clay) as well as compounds that may have been introduced after use (i.e. absorbed from the post depositional environment) has been assessed. One study [16] showed that ring pottery at 400e600  C for 30 min resulted in the loss of all oxygenated organic compounds, and most hydrocarbons from the claydin essence, giving pottery a clean slate for lipids prior to use. Another study showed that lipids extracted from soil in direct contact with pottery sherds (the dirt was actually scraped from the surface of the sherd) diered signicantly both qualitatively and quantitatively from those extracted from the interior of the sherd [13]. Also, in an early application of GC to archaeological materials, Condamin et al. [1] were able to show a signicant decrease in lipid concentration from the interior to the exterior portion of an amphora fragment, demonstrating that the former contents of the amphora, rather than the depositional environment, were likely to be the primary source of those lipids.
Table 5 Identications using Malaineys criteria and Marchbanks %S Method Malaineys criteria Marchbanks %S FMRS 6 Fish or corn Plant FMRS 7 None Plant FMRS 11 Beaver Fish FMRS 12 Beaver Fish

Similar tests need to be applied to rock materials. Whether or not a clean slate may be formed in rock materials has yet to be experimentally determined. It seems likely that as rocks are pre-heated for cooking some of the natural lipids would be destroyed, although the degree to which this would occur would probably depend on the type of rock and the cooking method employed. Furthermore, it is unclear how lipids are absorbed post-depositionally in rock. Future testing is needed to determine how heating aects the lipid content of rocks, and also what eects dierent rock porosities, and depositional environments have on lipid absorption and preservation.

8. Conclusions Although culturally introduced residues may be present in some of the FMRS samples, this study failed to demonstrate a signicant dierence between a group of cooking rocks and a group of o-site rocks. As a preliminary test of the application of fatty acid analysis to cooking rocks, this has been successful in demonstrating several things.

FMRS 24 Beaver Plant

FMRS 25 Beaver Land animal

FMRS 26 Beaver Fish

Rock 1 Beaver Fish

Rock 2 Beaver Plant

Rock 3 None Plant

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First, lipids can be extracted from the surface as well as the interior of andesitic rocks. The interior may be preferred if attempting to avoid some issues of contamination. In addition, it seems reasonable to decrease powdered rock samples to about 5 g. Second, because lipids are present in o-site rocks, and because rocks may contain naturally absorbed lipids in combinations that could lead to erroneous source identications, a reliable comparison of natural lipid concentrations and types should be made prior to interpreting lipids extracted from artifacts. Determining the concentration of lipids may be important for setting a baseline that could separate culturally introduced lipids from those present due to natural processes. In order to allow fatty acid concentrations to be compared between studies, absolute quantities rather than peak areas should be reported. Third, dierent identication criteria should also be assessed with regard to any naturally absorbed lipids. Research aimed at developing and rening methods for identifying absorbed lipid residues will continue to expand the utility of lipid analysis. Because they can avoid many of the problems associated with mixing and unpredictable degradation patterns, methods incorporating the detection of more specic biomarkers [4e 8,31] may have greater potential to supply meaningful data to archaeological analyses than broad based approaches using only GC of fatty acids. Because cooking rocks and ground stone tools have been used in food preparation for millennia, the amount of information that could potentially be gained from lipid analyses of these materials is great. Thus, continued eorts aimed at developing more informative and reliable techniques of lipid analysis of stone materials are worthwhile.

reviewers and were very helpful in improving the quality of this paper.

References
[1] J. Condamin, F. Formenti, M.O. Metais, M. Michel, P. Blond, The application of gas chromatography to the tracing of oil in ancient amphorae, Archaeometry 18 (1976) 195e201. [2] M.S. Copley, P.J. Rose, A. Clapham, D.N. Edwards, M.C. Horton, R.P. Evershed, Detection of palm fruit lipids in archaeological pottery from Qasr Ibrim, Egyptian Nubia, Proceedings of the Royal Society of London B 268 (2001) 593e597. [3] R.S. Dodd, Z.A. Rai, E. Zavarin, Eugene, Chemosystematic variation in acorn fatty acids of California Live Oaks (Quercus agrifolia and Q. wislizenii), Biochemical Systematics and Ecology 21 (1993) 279e285. [4] J.W. Eerkens, The origins of pottery among Late Prehistoric hunter-gatherers in California and the Western Great Basin, PhD dissertation, University of California, Santa Barbara, 2001. [5] J.W. Eerkens, The preservation and identication of Pinon resins by GC-MS in pottery from the western Great Basin, Archaeometry 44 (2002) 95e105. [6] R.P. Evershed, C. Heron, J. Goad, Analysis of organic residues of archaeological origin by high-temperature gas chromatography and gas chromatography-mass spectrometry, Analyst 115 (1990) 1339e1342. [7] R.P. Evershed, C. Heron, J.L. Goad, Epicuticular wax components preserved in potsherds as chemical indicators of leafy vegetables in ancient diets, Antiquity 65 (1991) 540e544. [8] R.P. Evershed, Biomolecular archaeology and lipids, World Archaeology 25 (1993) 74e93. [9] G.J. Farris, Aboriginal use of pine nuts in California: an ethnological, nutritional, and archaeological investigation into the uses of the seeds of Pinus lambertiana Dougl. and Pinus sabiniana Dougl. by the Indians of Northern California, PhD dissertation, University of California, Davis, 1982. [10] N. Garnier, P. Richardin, V. Cheynier, M. Regert, Characterization of thermally assisted hydrolysis and methylation products of polyphenols from modern and archaeological vine derivatives using gas chromatography-mass spectrometry, Analytica Chimica Acta 493 (2003) 137e157. [11] F. Gunstone, Fatty acid and lipid chemistry, Aspen Publishers Inc, Maryland, 1999. [12] J.L. Harwood, N.J. Russell, Lipids in plants and microbes, Allen and Unwin Inc, Mass, 1984. [13] C. Heron, R.P. Evershed, J.L. Goad, Eects of migration of soil lipids on organic residues associated with buried potsherds, Journal of Archaeological Science 18 (1991) 641e659. [14] G. Hillman, S. Wales, F. McLaren, J. Evans, A. Butler, Identifying problematic remains of ancient plant foods: a comparison of the role of chemical, histological, and morphological criteria, World Archaeology 25 (1993) 94e121. [15] M. Kates, Techniques of lipidology: isolation, analysis and identication of lipids, second ed., Elsevier, New York, 1986. [16] J.S. Johnson, J. Clark, S. Miller-Antonio, D. Robins, M.B. Schier, J.M. Skibo, Eects of ring temperature on the fate of naturally occurring organic matter in clays, Journal of Archaeological Science 15 (1988) 403e414. [17] R. Levy, Eastern Miwok, in: R.F. Heizer (Ed.), Handbook of North American Indians, vol. 8, Smithsonian Institution, Washington, 1978, pp. 398e413. [18] M.E. Malainey, R. Przybylski, B.L. Sherri, The fatty acid composition of native food plants and animals of Western Canada, Journal of Archaeological Science 26 (1999) 83e94.

Acknowledgements This research was supported by a Fall 2002, Graduate Research and Creativity Grant from the CSU, Chico Research Foundation. Thanks to Dr Sam Beattie (California State University, Chico) for providing access to the soil lipid labs, assistance with instrumental analysis, and support during the early phase of this study. Thanks also to Dr Jelmer Eerkens (University of California, Davis) for providing valuable advice and critiques on two earlier drafts. Thanks to Dr Greg White (Director of the Archaeological Research Program at CSU, Chico) for having the forethought to save the Fort Mountian Rockshelter cooking rocks for future analysis, as well as for his advice and availability throughout the research process. Thanks to Dr Frank Bayham (California State University, Chico) for providing comments and suggestions on an earlier draft. Useful critiques were provided by three anonymous

T. Buonasera / Journal of Archaeological Science 32 (2005) 957e965 [19] M.E. Malainey, R. Przybylski, B.L. Sherri, The eects of thermal and oxidative decomposition on the fatty acid composition of food plants and animals of Western Canada: implications for the identication of archaeological vessel residues, Journal of Archaeological Science 26 (1999) 95e103. [20] M.E. Malainey, R. Przybylski, B.L. Sherri, Identifying the former contents of late precontact period pottery vessels from Western Canada using gas chromatography, Journal of Archaeological Science 26 (1999) 425e438. [21] M.L. Marchbanks, Lipid analysis in archaeology: an initial study of ceramics and subsistence at the George C. Davis site, MA thesis, University of Texas, Austin, 1989. [22] M. Marchbanks, Organic residue analysis, in: M.B. Collins, B. Ellis, C. Dodt-Ellis (Eds.), Excavations at the Camp-Pearl Wheat Site (41KR243): An Early Archaic Campsite on Town Creek, Kerr, County, Texas, Studies in archaeology 6, University of Texas, Austin, 1990. [23] J. McMurry, Organic chemistry, third ed., Brooks/Cole Publishing Company, California, 1992. [24] C.H. Merriam, The acorn, a possibly neglected source of food, National Geographic Magazine 34 (1918) 129e137. [25] K. ODonoghue, A. Clapham, R.P. Evershed, T.A. Brown, Remarkable preservation of biomolecules in ancient radish seeds, Proceedings of the Royal Society of London B 263 (1996) 541e547. [26] D.H. ORourke, M.G. Hayes, S.W. Carlyle, Ancient DNA studies in physical anthropology, Annual Review of Anthropology 29 (2000) 217e242.

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[27] M. Patrick, A.J. de Koning, A.B. Smith, Gas liquid chromatographic analysis of fatty acids in food residues from ceramics found in the Southwestern Cape, South Africa, Archaeometry 27 (1985) 231e236. [28] M.J. Quigg, M.E. Malainey, R. Przybylski, G. Monks, No bones about it: using lipid analysis of burned rock and groundstone residues to examine Late Archaic subsistence practices in South Texas, Plains Anthropologist 46 (2001) 283e303. [29] O.C. Shanks, R. Bonnichsen, A.T. Vella, W. Ream, Recovery of protein and DNA trapped in stone tool microcracks, Journal of Archaeological Science 28 (2001) 965e972. [30] J.M. Skibo, Pottery function: a use alteration perspective, Plenum Press, New York, 1992. [31] J.M. Skibo, M. Deal, Pottery function and organic residue: an appraisal, in: C. Yeung, W.B. Li (Eds.), Conference Papers on Archaeology in South-east Asia, University Museum and Art Gallery, University of Hong Kong, 1995, pp. 319e329. [32] K.D. Thomas, Molecular biology and archaeology: a prospectus for inter-disciplinary research, World Archaeology 25 (1993) 1e 17. [33] G. White, Archaeological investigations at Fort Mountain Rockshelter (Ca-Cal-991): a Late Prehistoric habitation site in central Calaveras County, California, report on le at the Northcentral Information Center of the California Archaeological Inventory, Sacramento State University, 1988. [34] J.H. Zar, Biostatistical analysis, fourth ed., Prentice Hall, New Jersey, 1999, pp. 146e150.