Journal of Archaeological Science (1999) 26, 8394 Article No. jasc.1998.0305, available online at http://www.idealibrary.

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The Fatty Acid Composition of Native Food Plants and Animals of Western Canada
M. E. Malainey
Department of Anthropology, University of Manitoba, Winnipeg, MB R3T 5V5, Canada

R. Przybylski
Department of Foods and Nutrition, University of Manitoba, Winnipeg, MB R3T 2N2, Canada

B. L. Sherri
Department of Geological Sciences, University of Manitoba, Winnipeg, MB R3T 2N2, Canada (Received 4 November 1997, revised manuscript accepted 10 April 1998)
In order to facilitate the identication of residues absorbed into the walls of Late Precontact vessels, a collection of more than 130 native food plants and animals from Western Canada was established. The fatty acid composition of uncooked plant and animal samples were determined using gas chromatography. Hierarchical cluster and principal component analyses show similarities in fatty acid composition of these foods which generally correspond to meaningful categories. Large mammal fat, large herbivore meat, sh, plant greens, roots and berries/seeds can be dierentiated. The fatty acid composition of small mammals was more similar to plants than to large mammals.
 1999 Academic Press

Keywords: NATIVE FOOD, FATTY ACIDS, COMPOSITION, WESTERN CANADA, GAS CHROMATOGRAPHY.

Introduction
ottery was commonly manufactured during the Late Precontact Period (2000 to European contact) by the hunting and gathering peoples of Western Canada. Although hundreds of vessels are found in sites dating to this period, the introduction of the European copper kettle in the 18th century led to the rapid disappearance of the Native pottery technology. Relatively little is known about how these conoidal and globular vessels were used, although the presence of carbonized residues suggest many functioned as cooking pots (Malainey, 1995). The identication of residues absorbed in the walls of these vessels may provide information about pottery use and expand our understanding of cooking practices, diet and general subsistence strategies beyond that derivable from traditional sources, such as faunal remains and tool assemblages. Certain aspects of the analysis of absorbed vessel residues remain unchanged since it was rst accomplished by Condamin et al. (1976); in particular, the
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use of gas chromatography to characterize the lipid component of the residue. In the past 10 years, vessel residue analysts have turned from GC alone to gas chromatography combined with mass spectroscopy (GC/MS) (Deal & Silk, 1988; Deal, 1990; Heron & Pollard, 1988; Heron, 1989; Evershed, Heron & Goad, 1990, 1991; Gerhardt, Searles & Biers, 1990; Heron, Evershed & Goad, 1991; Evershed et al., 1992; Skibo, 1992; Heron & Evershed, 1993; Charters et al., 1995). The higher cost of GC/MS and more limited access to these instruments are serious disadvantages. Therefore, a method of using GC alone to identify vessel residues would be benecial. Lipids used in residue characterization include sterols, waxes and fatty acids. Sterols are biological regulators which can be used to discriminate animal, plant and animal/plant combinations in the vessel residues (Evershed et al., 1992; Evershed, 1993). The presence of a particular wax, a long-chain alkyl compound, enabled the very precise identication of the former contents of a vessel as plants of the Brassica family (Evershed, Heron & Goad, 1991). One major
 1999 Academic Press

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advantage of employing GC/MS for vessel residue analysis is its capability to detect and identify traces of sterols and waxes. Fatty acids are the major constituents of fats and oils and occur in nature as triacylglycerols, consisting of three fatty acids attached to a glycerol molecule by ester-linkages. Unsaturated fatty acids decompose more readily than saturated fatty acids, sterols and waxes. In the course of decomposition, simple addition reactions may occur at points of unsaturation (Solomons, 1980) or peroxidation may lead to the formation of a variety of volatile and non-volatile end products which continue to degrade (Frankel, 1991). Determining the composition of uncooked plants and animals is an important rst step in the identication of archaeological residues; but because of fatty acid decomposition, direct comparisons between uncooked plants and animals and archaeological residues are not possible. In this study, food plants and animals native to Western Canada were collected and their fatty acid compositions were determined using GC.

Experimental Methods
Selection and processing of food samples The compilation of Manitoba food plants by Shay (1980) and ethnographic accounts of cooking practices determined which samples to include in this study. The edible plants identied are consistent with those recognized in other sources (Gilmore, 1919; Gaertner, 1967; Kindscher, 1987; Young, 1993). Over 130 plants, mammals, sh and birds used by natives as food were gathered, mostly from the plains, parkland and southern boreal forest of Manitoba. For each plant variety collected, one specimen was pressed and retained for identication; the others were cleaned with distilled water and then air dried at room temperature. In cases where there were seasonal dierences in the exploitation of the greens, seeds and roots of a plant, the fatty acid composition of each part was analysed separately. The collection includes berry samples from Wood Mountain, Saskatchewan. Native varieties of corn, squash, sunower and red bean were obtained from the University of North Dakota. Samples of black bear, deer, small mammals, grouse and catsh were donated to the study; whereas samples of bison meat, bone marrow and all other sh were purchased commercially. Researchers have found that the fatty acid composition of some domestic oilseeds are aected by variety, planting conditions and climatic conditions during the course of the growing season (Sheppard, Iverson & Weihrauch, 1978). Most wild plant specimens were collected in 1994; adequate levels of rain and sun produced favourable growing conditions. Plant specimens were collected as close as possible to their optimal time of utilization.

Lipid extraction of food samples Immediately prior to lipid extraction, dried plant samples were ground with an electric coee grinder while esh samples were nely chopped. Lipids were extracted by homogenization (2 2 min) of 10 g of the sample in a 2:1 v/v mixture of chloroform and methanol (2 50 ml) (Folch, Lees & Stanley, 1957). Solids and the non-lipid component were removed by ltering the solvent/lipid mixture into a separatory funnel followed by a distilled water wash (33 ml). After separation, the lower lipid-chloroform layer was collected and solvents were removed by evaporation. After transferring the sample to a small, pre-weighed vial, the lipid was dried with benzene under nitrogen until its weight remained constant. Lipid samples were stored in chloroform-methanol (2:1 v/v), ushed with nitrogen in a vial sealed with a teon-lined cap and placed in a 20 C freezer. Fatty acids were transesteried by treating 200 mg of the total lipid extract dissolved in 1 ml of petroleum ether with 12 ml of 05 N anhydrous hydrochloric acid in methanol (6570 C, 60 min). Fatty acids which occur in the sample as di- or triglycerides are detached from the glycerol molecule and converted to methyl esters. After cooling to room temperature, 5 ml of iso-octane was added, followed by 6 ml of distilled water and the sample was mixed. When the upper layer had clearly separated, 25 ml of the fatty acid methyl ester solution was placed in a vial and crimped with an aluminum cap with a teon lining. Gas chromatography analysis parameters The separation was performed on a Hewlett-Packard 5890 gas chromatograph tted with a ame ionization detector connected to a Hewlett-Packard 3390 computing integrator. Samples were separated using a Supelcowax 10 fused silica capillary column (30 m 025 mm I.D., Supelco, Oakville, ON). An autosampler injected a 1 l sample using a split injection system with ratio set at 1:80. Hydrogen was used as the carrier gas at a linear velocity of 40 cm/s. The column temperature was programmed from 190 to 235 C at 2 C per min; lower and upper temperatures were held for 3 and 10 min, respectively. Peaks were identied through comparisons with several external qualitative standards (NuCheck Prep, Elysian, MN). Using this procedure, fatty acids are detectable to the nanogram level. Statistical analysis In this study, the total fatty acid compositions of modern food samples were characterized by hierarchical cluster and principal component analyses. These methods are able to process the high number of variables resulting from fatty acid analysis. Furthermore, the results of the hierarchical cluster analysis can be independently veried by the principal component

Native Food Plants and Animals of Western Canada

Average root-mean-square distance 0.9 0.8 0.7 0.6 0.5 0.4

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1.3

1.2

1.1

1.0

0.3

0.2

0.1

0 I II Deer fat III IV

Large mammal A V VI VII B VIII IX X Mixed Fish Berries + nuts Mixed Seeds + berries

Squirrel Roots

XI Greens

XII XIII

Berries

Roots

XIV

Greens XVRoots Grouse Cattail seed

Figure 1. Dendrogram of the hierarchical cluster analysis of the fatty acid compositions with subclusters identied.

analysis. The SAS hierarchical cluster analysis program, PROC CLUSTER, set to determine average linkage, was used to identify relationships among the fatty acid compositions of the modern food samples. The clusters generated provided a framework for presenting the compositional data. The data was also analysed with principal component analysis, a multivariate technique, using the SAS program, PROC PRIN.

Results
Hierarchical cluster analysis Hierarchical cluster analysis of the fatty acid composition of food samples results in the formation of three major clusters, A, B and C, each consisting of a number of subclusters (Figure 1). The three large clusters reect broad similarities in the fatty acid composition of its individual members. Cluster A contains all the large mammal and sh samples. Cluster B contains primarily plant seeds and berries, but some plant roots and three small mammals are also

included. The samples in cluster C are mainly plant greens and roots, but rosehips and bearberries appear as well. Four samples, deer fat, cattail seeds, grouse and squirrel, did not readily cluster with any other samples. Tight clusters indicate strong similarities in the fatty acid composition of the samples; loose linkages signify lower degrees of correspondence. The discriminating characteristics of the food sample clusters are summarized in Table 1. The fatty acid composition of the members of the major subclusters, I to XV, and the four samples which did not readily link with other samples, deer fat, squirrel, grouse and cattail seed, are described below. The fatty acid compositions of the food plant and animals samples are presented in Tables 39. In cases where more than one specimen was collected at the same stage of maturation, an average composition is given. Cluster A, the large mammal and sh cluster, includes 11 samples. It consists of four subclusters, I to IV, and the deer fat sample which does not readily cluster. The samples in subcluster I, bear fat and cow bone marrow, have levels of C18:1 isomers averaging 57%. The dierent location of the single point of

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Table 1. Summary of average fatty acid compositions of food sample clusters Cluster Subcluster A I II III IV V VI B VII VIII IX X XI C XII XIII XIV XV

Type C16:0 C18:0 C18:1 C18:2 C18:3 VLCS VLCP

Mammal Large fat and herbivore marrow meat 1990 706 5677 701 068 016 018 1939 2035 3579 893 261 032 413

Fish

Fish

Berries Seeds and and nuts Mixed berries Roots Seeds Mixed Greens Berries Roots Greens Roots 373 173 5400 3785 093 071 002 1206 236 3529 3583 366 446 064 748 258 2912 5469 151 298 042 1998 259 655 4874 724 850 039 752 355 1002 6414 549 519 054 1033 243 1562 3924 1977 373 088 1871 248 503 1882 3508 677 022 347 134 1495 2908 3975 910 014 2268 315 1212 2624 964 1532 030 2419 366 405 1615 1788 1868 018 1871 594 334 1561 342 4336 040

1607 1410 387 278 1828 3196 291 404 439 383 023 015 3927 2324

VLCS Very Long Chain (C20, C22 and C24) Saturated Fatty Acids. VLCP Very Long Chain (C20, C22 and C24) Polyunsaturated Fatty Acids. Table 2. Variation in modern references explained by principal components 1 to 11 Principal component PRIN1 PRIN2 PRIN3 PRIN4 PRIN5 PRIN6 PRIN7 PRIN8 PRIN9 PRIN10 PRIN11 Eigenvalue 801 425 312 250 187 172 145 144 115 106 103 Fatty acids with high component loadings Polyunsaturates Medium chain; C18:1; C18:2 Medium chain C17:0; C19:0 Unsaturates C16:0; C17:0; C18:0 C18:3 6, C18:4 monounsaturates C17:1; C20:2; C20:4 6 C20:1; C22:1 C14:1; C18:1 11; C20:4 6 Percentage variation 2224 1180 866 693 519 478 403 399 319 293 286 Cumulative variation 2224 3404 4271 4964 5484 5961 6364 6763 7082 7375 7661

unsaturation in the two C18:1 fatty acid isomers, C18:1 9 and C18:1 11, results in slightly dierent retention times and peak separation; the value presented represents the sum of the two peaks. The average levels of C16:0 (20%), as well as, C18:0 and C18:2 (both 7%) are also signicant. Subcluster II includes the meat of two herbivores: bison and deer. The average fatty acid composition of these samples is characterized by high levels of C18:1 isomers (36%), C18:0 (20%), C16:0 (19%) and C18:2 (9%). Subclusters I and II link and are then joined by deer fat. Deer fat diers from the other samples in terms of its higher level of C18:0 (39%) and lower level of C18:1 isomers (24%). While diet can alter the fatty acid composition of animal tissue, these changes tend to impact polyunsaturated fatty acids and absolute levels are aected more than relative values. There are broad similarities in the composition of large herbivore products. The fatty acid composition of bison and deer meat are similar to values reported for the meat of moose, caribou, domestic oxen, Cape bualo and several

dierent species of African antelope (Crawford et al., 1970; Appavoo, Kubow & Kuhnlein, 1991). The fatty acid composition of the subcutaneous fat of white tail deer from southeast Manitoba is very similar to that of the wild African eland (Crawford et al., 1970). All of the sh samples appear in subclusters III and IV. Subcluster III includes pickerel, perch, catsh and smoked trout; subcluster IV contains whitesh and smoked goldeye. The division between the two subclusters reects higher levels of C18:1 isomers and lower levels of C22:6 in whitesh and smoked goldeye, as compared to the other sh samples. The average fatty acid composition of the sh samples is characterized by high levels of both very long chain (20 or more carbons in length) polyunsaturated fatty acids (VLCP) 39% for subcluster III and 23% for subcluster IV and C18:1 isomers 18% for subcluster III and 32% for subcluster IV. The average level of C16:0 in the subclusters is about 15%; the average level of C16:1 isomers is 8% for subcluster III and 12% for subcluster IV. Smoking is a method of preservation which does not aect the fatty acid composition of the sh

Native Food Plants and Animals of Western Canada

Table 3. The fatty acid composition of mammal and bird samples Sample Type Subcluster C14:0 C16:0 C16:1 C17:0 C18:0 C18:1 C18:2 C18:3 3 C20:4 6 C20:5 Others bear fat I 112 1816 220 035 589 6057 854 089 004 000 224 cow marrow I 377 2165 489 051 822 5298 547 048 005 000 198 bison meat II 264 1880 490 120 2234 3807 650 060 165 000 329 deer meat II 141 1998 182 107 1835 3351 1137 463 351 175 261 deer fat N/A 232 2260 134 193 3878 2370 346 264 010 000 088 beaver meat VI 083 1743 457 026 641 3831 2407 287 078 118 329 muskrat meat X 087 1686 148 104 608 1716 3483 1824 051 011 228 squirrel meat N/A 022 977 067 032 798 1871 4366 075 826 040 170 grouse meat N/A 055 1577 311 021 1325 1928 1795 686 992 659 307

87

Table 4. The fatty acid composition of sh samples Sample Type Subcluster C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 3 C18:4 C20:1 C20:4 6 C20:4 3 C20:5 C22:3 C22:4 C22:5 C22:6 Others catsh sh III 229 1600 812 452 2027 345 639 133 079 601 000 930 136 000 350 830 838 pickerel sh III 140 1712 763 367 1710 202 419 119 054 364 151 869 159 164 303 2086 419 perch sh III 102 1838 734 370 1533 209 300 067 035 539 090 808 157 173 315 2372 357 trout smk sh III 339 1277 695 358 2042 407 398 318 081 312 393 556 154 247 412 1510 503 whitesh sh IV 319 1386 1396 295 2858 226 198 157 122 195 097 938 054 069 352 918 421 goldeye smk sh IV 251 1434 1040 260 3534 581 568 171 028 223 296 616 042 012 278 324 341

samples. For a complete review of the factors which inuence the lipid composition of freshwater sh, readers should consult Henderson & Tocher (1987). Cluster B consists of 46 samples divided into six subclusters, V-X, of seeds, berries, roots and small mammals. Squirrel meat also appears in cluster B, but it is only loosely linked with the other samples. Subcluster V is a small cluster (N =6) of chokecherry, pincherry and wild hazelnuts in two minor clusters. The average fatty acid composition of these samples features very high levels of C18:1 isomers (54%) and C18:2 (38%). Subcluster VI contains samples of dock (N =3) and knotweed (N =1) seeds, two varieties of mushroom,

acorn and beaver meat in two minor clusters. The average fatty acid composition of the samples in subcluster VI contain high levels of C18:2 (36%) and C18:1 isomers (35%). The average level of C16:0 is 12%. The compositions of the mushrooms and beaver dier from other samples with respect to their higher levels of C16:1 isomers. The alpha linolenic fatty acid, C18:3 3 is virtually absent from mushrooms. The level of C18:2 in beaver meat is more than 10% lower than the mean for this subcluster. Subcluster VII consists of a tight cluster of eight seeds and berries, including most of the Native cultigens: Mandan Sweet Corn, Assiniboin Flint Corn, Arikara winter squash and Mandan sunower. Wild

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Table 5. The fatty acid composition of berry samples chokecherry V (N =4) 405 200 5293 3825 105 024 018 045 029 056 050 071 395 488 008 004 002 058 010 017 pincherry V&VII (N =2) 215 083 4310 5219 065 013 018 019 004 056 022 016 746 805 031 004 003 026 001 009 gooseberry X (N =2) 842 179 1289 3644 2241 126 017 068 197 1399 saskatoon VII (N =2) rosehips XII (N =4) 173 064 236 463 112 091 009 103 087 125 bearberry XII (N =2) 232 098 1252 2099 4060 040 047 664 1385 124 269 020 199 633 640 011 002 372 758 042 haw red blueberry juniper silverberry thorn osier X 688 171 1719 4170 2876 132 025 046 056 105 X 868 339 1484 3643 2066 057 114 203 095 9992 XIV 2088 324 781 1581 1838 408 012 1770 753 446 VII 716 196 2345 5687 213 163 069 065 100 445 XII 2660 136 1647 3809 1476 016 024 014 044 174

Sample Subcluster C16:0 C18:0 C18:1 C18:2 C18:3 3 C20:0 C20:1 C22:0 C24:0 Others
1 2

023 761 033 192 221 2543 110 5236 528 264 040 269 001 103 096 357 009 130 2381 148

023 405 018 152 192 1616 034 3313 052 3933 033 132 008 066 069 107 024 084 005 194

C18:3 6=851 168%; C18:4=343 033%. C12:0=386%; C20:2=335%; C22:0=203%. Table 6. The fatty acid composition of nut, bean and mushroom samples acorn nut VI 913 038 030 182 3774 4110 579 059 074 085 154 hazelnut nut V 387 037 007 146 6523 2762 053 024 008 017 036 Native redbean bean XI 1371 008 019 232 1088 3474 3535 045 074 093 061 vetchling pods VIII 1469 271 042 223 948 5409 954 107 134 218 225 Armillaria mushroom VI 1001 661 003 231 4550 3071 000 030 026 057 360 Leccinum mushroom VI 1285 180 011 090 3976 4065 014 014 021 018 324

Sample Type Subcluster C16:0 C16:1 C17:0 C18:0 C18:1 C18:2 C18:3 3 C20:0 C22:0 C24:0 Others

varieties in this subcluster include bulrush, saskatoon and hawthorn, which later links with a pincherry sample. The average fatty acid composition of these samples is characterized by very high levels of C18:2 (55%) and C18:1 isomers (29%), which is typical for many vegetable oils. The pincherry sample diers from the others in terms of its very low level of C16:0 and C18:1 isomer level, almost 9% higher than the subcluster mean. Most berries and the bulrush seed have higher levels of C18:3 3 and C24:0 than other samples in this cluster, saskatoon and bulrush seed have higher levels of C22:0. The level of C18:0 in sunower and winter squash is somewhat higher than in the other samples. The fatty acid composition of Native corn diered from commercial corn oil (Sheppard, Iverson & Weihrauch, 1978) in that the average level of C18:1 isomers was 5% higher and C18:2 was 5% lower in the Native cultigens. The values obtained for Mandan sunower corresponded well to average values for

commercial sunower oil (Sheppard, Iverson & Weihrauch, 1978). Subcluster VIII consists of 10 plant roots, including false solomons seal, cow parsnip, Jerusalem artichoke, wild onion, prairie turnip, tiger lily, water parsnip and wild calla, as well as a sample of vetchling pods in two minor clusters. The average fatty acid composition of the samples is characterized by very high levels of C18:2 (49%) and C16:0 (20%). Some roots contain levels of medium chain or very long chain saturated (VLCS) fatty acids which are signicantly higher than the mean for the entire cluster. The level of C12:0 in Jerusalem artichoke is almost six times higher than the mean; C15:0 levels are signicantly higher in both Jerusalem artichoke and cow parsnip. The level of C22:0 is 6% in prairie turnip, double the mean of 3%. The level of C24:0 is over 12% in tiger lily root and approaches 8% in prairie turnip, whereas the subcluster mean is 4%.

Native Food Plants and Animals of Western Canada

Table 7. The fatty acid composition of plant seed samples marshelder IX 901 132 029 629 595 5970 012 513 342 047 000 236 106 095 392 arrowgrass IX (N =2) 678 094 113 053 032 016 218 008 1206 153 6636 608 021 027 567 349 166 065 017 002 000 168 081 000 107 035 074 035 dock1 VI(N =3), X, XIII 1256 214 125 099 022 006 182 033 2431 587 3485 688 000 1030 758 164 062 094 038 124 112 219 075 002 004 613 420 211 081 knotweed IX 1156 081 034 161 3557 3670 000 588 070 141 092 134 000 122 195 goosefoot2 XIII(N =2), X 1337 194 026 177 1537 3872 002 1613 139 107 109 251 010 225 323 476 173 006 026 169 1266 003 528 048 045 075 093 009 189 059 Native corn3 VII (N =2) 1061 016 004 001 009 001 217 030 3216 049 5240 025 000 116 004 054 005 032 001 000 018 001 000 023 002 013 001 Native sunower VII 572 002 009 446 3160 5571 000 013 041 021 000 124 000 030 012 Native squash VII 1015 011 015 741 2897 5181 000 018 053 008 000 014 000 012 034 bulrush VII 588 020 020 054 2501 6040 000 316 071 049 007 152 000 105 083

89

Sample Subcluster C16:0 C16:1 C17:0 C18:0 C18:1 C18:2 C19:0 C18:3 3 C20:0 C20:1 C22:1 C22:0 C22:2 C24:0 Others
1 2 3

cattail N/A 916 044 2025 334 458 1113 2221 311 296 027 063 1355 028 611 199

N =5. N =3. Mandan int corn and Assiniboin sweet corn.

Subcluster IX is a small cluster of marsh-elder (N =1) and arrow-grass (N =2) seeds. The seeds have an average fatty acid composition which is extremely high in C18:2 (64%), with lower levels of C18:1 isomers (10%) and C16:0 (8%). The levels of other fatty acids vary widely between these three samples. Subcluster X contains mainly plant material, dock and lambs-quarters seeds and gooseberry, blueberry and juniper berries, but muskrat meat also joined this subcluster. This subcluster contains a number of minor clusters and the average fatty acid compositions are quite variable. In general, the average level of C18:2 is high (39%) and signicant amounts of C18:3 3 (20%), C18:1 isomers (16%) and C16:0 (10%) are present. Juniper berries dier from the other samples on the basis of its higher levels of C12:0 and C14:0. The level of C18:0 is higher in muskrat while the two gooseberry samples have high levels of C18:3 6 and C18:4. The levels of very long chain saturated and mono-unsaturated fatty acids vary widely between samples. Squirrel meat likely appears in cluster B because of its high level of C18:2 (44%), but it does not closely resemble any of the samples. Cluster C consists of ve subclusters of plants, XI to XV, and two samples that did not readily cluster (grouse esh and cattail seed). Subcluster XI is dominated by plant greens, specically false solomons seal, reweed, golden rod, violet, dock and goosefoot which form two minor clusters. In total, there are 24 plant samples, 22 of which are plant greens, one plant seed and one plant root. The average fatty acid compositions are characterized by high levels of C18:3 3 (35%) and C16:0 (19%). The two minor clusters within subcluster XI may reect dierences in the levels of

branched C16:0 (brC16:0), C16:1 isomers and C18:2 in the samples. False solomons seal greens, reweed root and the Native domesticate, Hidatsa red beans, all have relatively low levels of C16:1 isomers; false solomons seal and violet greens both have relatively high levels of C14:1. Stinging nettle and one sample of sarsaparilla greens were the last to join established clusters. The high C24:0 level in the stinging nettle and the levels of C18:2, C18:3 3 and C24:1 in the sarsaparilla greens distinguish them from the other samples. Subcluster XII consists exclusively of rosehips and bearberries. The average fatty acid composition of these samples is characterized by high levels of C18:3 3 (40%), C18:2 (29%) and C18:1 isomers (15%). Both bearberry samples have higher levels of C22:0 and C24:0. One of these has lower levels of C18:1 isomers and C18:2 than the other samples and is the last to join the cluster. Subcluster XIII is dominated by plant root samples, including bulrush, cattail, water parsnip, giant reed, reweed, sarsaparilla, water-hore hound, wound wort, ostrich fern and Jerusalem artichoke. Nineteen of the 24 samples in subcluster XIII are roots; red osier berries, mares tail and chickweed greens, and lambsquarter and dock seeds also appear. The members of this subcluster are rather loosely linked, consisting of a number of minor clusters and the fatty acid composition of samples is variable. Average levels of C18:2 (26%) and C16:0 (23%) are quite high, but there are signicant amounts of C18:1 (12%) and C18:3 3 (10%), as well. The levels of medium chain fatty acids (C12:0, C14:0 and C15:0) are much higher than average in water parsnip and Jerusalem artichoke. Levels of VLCS (C20:0, C22:0 and C24:0) are very high in some samples, but there is considerable variability.

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Table 8. The fatty acid composition of plant root samples false solomons seal VIII (N =2) 023 014 069 007 2324 115 152 100 000 279 032 516 220 4741 679 602 170 164 050 347 245 261 161 301 218 025 025 197 054 reweed1 water parsnip2 cow parsnip VIII (N =2) 035 120 1850 029 004 184 489 5636 614 191 092 209 268 051 232 Jerusalem artichoke3 VIII, XIII 110 227 354 007 276 042 088 1274 131 028 006 173 033 052 206 cattail4 XV (N =2), XIII 013 004 097 014 1815 394 184 090 000 491 179 397 264 2151 1026 465 226 1003 259 074 021 1130 263 1681 477 005 008 478 211 bulrush XIII (N =4) 011 109 2579 233 015 190 1364 2126 821 204 063 676 1299 044 266 giant reed XIII (N =2)

Sample Subcluster C12:0 C15:0 C16:0 C16:1 C16:2 C18:0 C18:1 C18:2 C18:3 3 C20:0 C20:1 C22:0 C24:0 C24:1 Others

VIII (N =3), XI XIII(N =2), VIII 002 036 2687 145 040 361 1475 2137 1670 342 064 197 157 085 600 004 010 585 074 044 068 1040 073 854 231 042 039 060 123 324 360 295 449 428 1922 309 117 154 110 150 437 060 696 251 3707 931 572 085 308 058 084 044 427 171 229 145 000 581 371

001 311 015 403 300 1845 001 069 006 315 013 172 155 496 021 4177 015 826 092 102 023 062 041 382 123 220 071 166 076 436

013 000 082 034 025 327 2019 457 151 040 012 029 014 019 143 268 025 099 786 318 723 3157 640 196 682 091 053 1283 221 047 708 277 198 336 062 478 439 027 042 010 013 076 228 033

Sample Subcluster C12:0 C15:0 C16:0 C16:1 C16:2 C18:0 C18:1 C18:2 C18:3 3 C20:0 C20:1 C22:0 C24:0 C24:1 Others
1 2 3 4

sarsaparilla XIII, XIV (N =2) 041 031 037 009 2240 109 311 228 193 010 264 081 710 606 2642 484 1368 494 285 008 035 009 573 156 752 396 000 553 086

wild onion VIII 058 044 1975 290 017 403 1127 4004 533 283 058 356 354 159 339

tiger lily VIII 000 012 2033 139 028 246 776 4526 308 138 048 346 1262 000 139

wild calla VIII 000 034 2510 253 169 119 944 4246 1285 030 055 097 090 028 141

prairie tumip VIII 000 000 1837 000 000 383 426 4813 881 203 000 604 762 000 091

burreed XIII 027 069 1974 368 000 303 1998 2011 373 433 111 523 1464 000 348

arrow head XIV 012 007 2030 366 048 717 299 1841 1322 135 017 259 2459 027 275

wound wort XIII 000 020 2120 632 000 281 1063 2316 1217 273 037 744 883 085 328

ostrich fern XIII 010 087 3226 232 381 207 1000 3479 650 081 018 165 280 000 175

water-hore hound XIII 000 041 1909 087 614 474 1198 2639 1125 306 035 655 517 123 276

N =4. N =3. N =2. N =3.

Subcluster XIV is dominated by plant greens, including cow parsnip, cattail, bulrush and golden rod. Ten of the 13 samples in this cluster are plant greens; the other samples are sarsaparilla and arrowhead roots and silverberry. The average fatty acid composition is characterized by a high level of C16:0 (25%) with moderate levels of VLCS (19%), C18:3 3 (18%) and C18:2 (16%). Branched C16:0 is common in this subcluster, appearing in all samples except silverberry. Subcluster XV contains two samples of cattail root. The average fatty acid composition of these samples is characterized by very high levels of VLCS; together, C24:0 (19%), C22:0 (13%) and C20:0 (12%) account for

almost 45% of all fatty acids. Signicant levels of C16:0 (19%) and C18:2 (16%) are also present. Grouse is likely included in cluster C because its fatty acid composition is characterized by a number of fatty acids (C16:0, C18:0, C18:1 isomers and C18:2) occurring at levels between 13 and 18%, rather than one or two dominant fatty acids. Cattail seed is only loosely linked to cluster C; it diers from the other samples on the basis of its high level of 17:0, 20%. Principal component analysis When all 37 fatty acids present in the data set are included in the analysis, each principal component

Native Food Plants and Animals of Western Canada

Table 9. The fatty acid composition of plant greens samples Sample Subcluster C14:0 C14:1 brC16:0 C16:0 C16:1 C16:2 C17:1 C18:0 C18:1 C18:2 C18:3 3 C20:0 C22:0 C24:0 C24:1 Others false solomons seal XI (N =5) 053 089 082 2055 422 087 036 236 338 2523 3248 112 221 248 007 243 021 092 043 284 176 023 023 042 165 393 514 022 028 048 007 051 golden rod1 XI (N =3), XIV 023 009 000 151 064 2115 498 852 258 200 032 000 269 055 202 068 1430 381 3630 625 242 073 185 034 290 169 097 021 189 114 reweed XI (N =4) 181 084 000 103 040 1823 189 779 210 225 018 006 013 286 072 295 079 1384 362 3810 217 299 091 219 077 313 098 056 055 221 060 cow parsnip XIV (N =3) 154 120 144 1825 549 097 332 279 370 2417 1951 221 336 720 007 484 137 081 099 397 359 092 252 047 182 340 345 059 066 140 012 412 bulrush XIV (N =3) 097 040 000 114 022 3051 299 844 097 110 012 012 021 281 029 507 039 1224 297 1613 455 279 029 527 131 974 181 004 008 367 079 cattail XIV (N =3) 120 079 000 101 033 2496 335 735 199 106 042 018 031 513 133 426 405 1191 342 1494 593 487 166 820 206 1148 356 000 345 087 violet

91

XI (N =2) 062 027 254 014 053 075 1884 151 624 107 197 021 009 013 213 030 664 005 2331 438 3119 078 058 040 130 097 284 339 000 119 042

Sample Subcluster C14:0 C14:1 brC16:0 C16:0 C16:1 C16:2 C17:1 C18:0 C18:1 C18:2 C18:3 3 C20:0 C22:0 C24:0 C24:1 Others
1

sarsaparilla XI (N =2) 113 090 000 178 014 1813 286 929 241 185 014 006 008 240 092 472 045 1134 496 4157 842 138 103 190 140 245 148 148 180 076 023

goosefoot XI (N =2) 085 050 000 190 039 1850 075 1072 253 227 061 189 045 141 037 643 084 1335 054 3489 083 190 037 262 016 240 030 000 091 080

dock XI (N =2) 050 021 002 002 181 033 1815 156 815 093 182 088 017 023 177 035 988 127 1408 245 3761 632 085 022 119 012 264 040 000 143 050

water-hore hound XI 064 000 000 1853 840 165 014 511 558 1035 3071 361 424 350 107 647

stinging nettle XI 041 000 245 1413 1014 171 000 214 297 1874 3085 298 319 951 000 079

chick weed XIII 152 000 000 1913 383 086 000 187 1102 3196 1505 102 192 418 011 123

mares tail XIII 085 000 174 2979 1003 107 000 439 1090 2126 929 250 328 328 000 034

N =4.

explains a relatively small part of the variation. The rst 11 principal components have eigenvalues greater than one and are considered signicant. Together they account for more than 76% of the total variation in the modern food plants and animals (Table 2). Principal component 1, which has high positive component loadings on polyunsaturated fatty acids with chainlengths of 20 or more carbons, accounts for 22% of the variation. Principal component 2, with high positive loadings on fatty acids with chain-lengths of 16 or fewer carbons and negative loadings on oleic and linoleic acids, explains almost 12% of the variation. Each of principal components 3 to 11 explain between 9 and 3% of the variation. The high loadings on polyunsaturated fatty acids in the rst principal component results in the separation of sh and grouse in

the plot of PRIN2 versus PRIN1 (Figure 2). Greens, roots, as well as berries, seeds and nuts, each form fairly tight clusters in the plot of PRIN2 versus PRIN3 (Figure 3). Reducing the number of variables under consideration would simplify the principal component analysis, but this action may introduce an unacceptable level of bias.

Discussion and Conclusions

When the fatty acid compositions of modern food plants and animals are subject to principal component and hierarchical cluster analysis, the resulting groupings generally correspond to divisions which exist in nature. Clear dierences in the fatty acid composition

92

M. E. Malainey et al.
6

PRIN2

Fat Large herbivore meat Fish Berry/seed/nut Greens Roots Small mammal meat Mushroom Bird

6 PRIN1

10

12

14

16

18

Figure 2. Plot of PRIN2 scores versus PRIN1 scores for the modern references.

of large mammal fat, large herbivore meat, sh, plant roots, greens and berries/seeds/nuts can be detected, but the fatty acid composition of medium-sized mammals meat closely resembles berries/seeds/nuts; their main dietary components. Samples in cluster A, the large mammal and sh cluster, have elevated levels of C16:0 and C18:1. Fat samples are high in C18:1 isomers, bison and deer meat have higher levels of C18:0 while sh have high levels of VLCP. The fatty acid composition of bison and deer products resemble those of other large herbivores from

North America (Appavoo, Kubow & Kuhnlein, 1991) and Africa (Crawford et al., 1970). Statistically signicant dierences in the fatty acid composition of plant roots, greens and berries/seeds/ nuts were based on dierences in the amounts of C18:2 and C18:3 3 in the samples. The berry, seed, nut and small mammal meat samples appearing in cluster B have high to extremely high levels of C18:2. Samples in subclusters VIII, IX and X have high to extremely high levels of C18:1 isomers, as well; squirrel meat has higher levels of VLCP than other samples in cluster B.

Native Food Plants and Animals of Western Canada

6

93

PRIN2

Fat Large herbivore meat Fish Berry/seed/nut Greens Roots Small mammal meat Mushroom Bird

2 PRIN3

10

Figure 3. Plot of PRIN2 scores versus PRIN3 scores for the modern references.

Modern food samples in cluster C are mainly plant roots and greens, but include rosehips and bearberries. These samples all have higher levels of C18:2 except for the berries, all samples have elevated levels of C16:0. Higher levels of C18:3 3 and/or VLCS are also common. Grouse meat diers from the other samples in cluster C because of its higher levels of C18:1 isomers and VLCP; cattail seeds have high levels of C17:0 and VLCS. The determination of the fatty acid composition of uncooked food plant and animal samples is an important rst step in the identication of vessel residues because it enables cooking experiments to be conducted on a representative sample of foods of similar

fatty acid compositions, rather than on all samples in the collection. The eects of thermal and oxidative degradation on meat, sh and plants, prepared alone or in combination, are reported in Malainey, Przybylski & Sherri (1999).

Acknowledgements
This paper is extracted from Malainey (1997). The research was carried out while MEM held a Social Science and Humanities Research Council of Canada doctoral fellowship. This research was supported by a Natural Science and Engineering Research Council

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spectrometry. In (W. R. Biers & P. E. McGovern, Eds) Organic Contents of Ancient Vessels: Materials Analysis and Archaeological Investigation Vol. 7. Philadelphia: MASCA, The University Museum of Archaeology and Anthropology, University of Pennsylvania, pp. 4150. Gilmore, M. (1919) Uses of plants by the Indians of the Missouri River region. Thirty-third Annual Report of the Bureau of American Ethnology. Washington: Government Printing Oce; reprinted 1991, University of Nebraska Press, Lincoln. Henderson, R. J. & Tocher, D. R. (1987). The lipid composition and biochemistry of freshwater sh. Progress in Lipid Research 26, 281347. Heron, C. P. (1989). The analysis of organic residues from archaeological ceramics. Ph.D. Dissertation, University of Wales. Heron, C., Evershed, R. P. & Goad, L.J. (1991). Eects of migration of soil lipids on organic residues associated with buried potsherds. Journal of Archaeological Science 18, 641659. Heron, C. & Evershed, R. P. (1993). The analysis of organic residues and the study of pottery use. In (M. B. Schier, Ed.) Archaeological Method and Theory 5. Tucson: University of Arizona Press, pp. 247284. Heron, C. & Pollard, A. M. (1988). The analysis of natural resinous materials from Roman amphoras. In (E. A. Slater & J. O. Tate, Eds) Science and Archaeology Glasgow 1987. Proceedings of a Conference on the Application of Scientic Techniques to Archaeology, Glasgow, 1987. Oxford: BAR British Series 196 (ii), pp. 429447. Kindscher, K. (1987) Edible Plants of the Prairie. Lawrence: University Press of Kansas. Malainey, M. E. (1995). Functional analysis of precontact pottery from West-central Canada. Manitoba Archaeological Journal 5, 6086. Malainey, M. E. (1997). The reconstruction and testing of subsistence and settlement strategies for the plains, parkland and southern boreal forest. Ph.D. Thesis, University of Manitoba. Malainey, M. E., Przybylski, R. & Sherri, B. L. (1999). The eects of thermal and oxidative degradation 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 (this issue). Shay, C. T. (1980). Food plants of Manitoba. In (L. Pettipas, Ed.) Directions in Manitoba Prehistory: Papers in Honour of Chris Vickers. Winipeg: Association of Manitoba Archaeologists and Manitoba Archaeological Society, pp. 233290. Sheppard, A. J., Iverson, J. L. & Weihrauch, J. L. (1978). Composition of selected dietary fats, oils, margarines, and butter. In (A. Kuksis, Ed.) Handbook of Lipid Research 1: Fatty Acids and Glycerides. New York: Plenum Press, pp. 341379. Skibo, J. M. (1992). Pottery Function: A Use-Alteration Perspective. New York: Plenum Press. Solomons, T. W. G. (1980). Organic Chemistry. Toronto: John Wiley & Sons. Young, K. (1993). Wild Seasons: Gathering and Cooking Wild Plants of the Great Plains. Lincoln: University of Nebraska Press.

University Research Fellowship and research grant to BLS and a Manitoba Heritage Grant to MEM. This project beneted from the advice and support of M. Latta, R. Zambiazi, G. Monks, B. Watts, E.L. Syms, M. Deal and two anonymous reviewers. The eorts of all those who aided in the collection of food plant and animal samples are gratefully acknowledged.

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