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Organic Chemistry of Embalming

Organic Chemistry of Embalming

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NATURE
|
VOL 413
|
25 OCTOBER 2001
|
www.nature.com
837
letters to nature
.................................................................
OrganicchemistryofembalmingagentsinPharaonicandGraeco-Romanmummies
Stephen A. Buckley & Richard P. Evershed
Organic Geochemistry Unit, Biogeochemistry Research Centre, School of Chemistry, University of Bristol, Cantock's Close, Bristol BS8 1TS, UK 
..............................................................................................................................................
Chemical treatments were an essential element of ancient Egyp-tian mummi®cation. Although the inorganic salt natron is recog-nized as having a central role as a desiccant
1
, without theapplication of organic preservatives the bodies would havedecomposed in the humid environment of the tombs
2
. Thenature of the organic treatments remains obscure, because theancient Egyptians left nowritten recordof the process. Secondary textual evidence for mummi®cation is provided by Herodotus
3
,Diodorus Siculus
4
, Strabo
5
and Pliny 
6
. The most importantaccount is that of Herodotus
3
(about450yr
BC)
, although archaeo-logical evidence shows that by this time the process had declinedsigni®cantly and the best results had been achieved centuriesbefore
7
.Hisaccountmentionsmyrrh,cassia,palmwine,`cedaroil'(still widely disputed
8±10
) and `gum'; however, it is vague withrespect to the speci®c natural products used. Here we report theresults of chemical investigations of a substantial collection of samples of tissues, wrappings and `resinous/bituminous' materi-als from provenanced and dated Egyptian mummies. We focusedon examples of the `classic' mummy-making culture of thePharaonic or dynastic period, from which we can begin to track the development of mummi®cation chronologically.
Studies of the organic materials used in mummi®cation werecarried out earlier in the last century (for example, see refs 11±13),but the analytical techniques available limited their ability toidentify aged, complex organic mixtures (that is, the `resins',spices, and so on). To gain a proper understanding of the mummi-®cation process and its development, it is vital to study securely provenanced and dated mummies using modern investigativechemical techniques. Only four such studies (which includes oneperformed in our own laboratory) have been carried out in recent
Table 1 Provenance and date of mummies, origin of balm samples and their chemical composition
MummyDate/age Provenance Sample location and description
#
Inferred components of embalming `resin'
I
Relativeabundance (%)**
...................................................................................................................................................................................................................................................................................................................................................................
Male adult* (`Khnumnakht')214711985±1795 yr
BC
(XII dynasty)Rifeh Resin/tissue/bandaging Fat/oil
e
Proteinaceous material90%10%Female adult²1909.5271650±1550 yr
BC
(XVII dynasty)Qurna, Thebes Resin/tissue from head of right tibia Fat/oil
c
Coniferous pitchBalsam (?)99%1% TraceChild (sex ?)²1909.5271650±1550 yr
BC
(XVII dynasty)Qurna, Thebes Unspeci®ed bone and cartilage Fat/oil
b
Coniferous pitchBalsam (?)82%1%16%Head (sex ?)³1976.159.2671550±1069 yr
BC
(XVIII±XX dynasties) Thebes Skin/resin from back of cranium Fat/oil
g
 A sugar/gumConiferous resinBalsam (?)95%0.3%2%2%Male adult§ (`Horemkenesi')Ha 73861069±945 yr
BC
(XXI dynasty)Deir el Bahri, Thebes Resinous material from left side of top of spineFat/oil
l
100%Female(?) adult
k
EA743031069±664 yr
BC
(XXI±XXV dynasties) Thebes? Resin from chest cavity Fat/oil
Coniferous resin/pitchBalsam (?)Beeswax
n,s
61%6%1.5%31%Female adult§ (`Neskhons')H 5062945±715 yr
BC
(XXII dynasty) Thebes Resin-soaked outer wrapping fromleft side of neck Plant oil
d
Coniferous resin Triterpenoid resin ?Balsam (?)Wax82%0.5%4%0.9%11%Male adult³ (`Pedeamun')1953.72664±404 yr
BC
(XXVI±XXVII dynasties) Thebes Resin from top of cranium Fat/oil
m
Coniferous resinBalsam (?)Beeswax
o,t
61%0.2%1.5%38%Female adult²1956.352332±30 yr
BC
(Ptolemaic) Thebes Resin attached to linen thread onright ankleFat/oil
Pistacia resinBalsam (?)Beeswax
p,u
6%6%0.4%87%Male adult (1)²1911.210130 yr
BC
to
AD
395(Roman)Hawara Resin-soaked outer wrapping belowright scapulaFat/oil
i
Coniferous resinBalsam (?)Beeswax
q,v
78%16% Trace6%Male adult (2)²1911.210230 yr
BC
to
AD
395(Roman)Hawara Resin from side/base of left foot Fat/oil
a
Coniferous resinBalsam (?)Beeswax
r,w
35%37%0.2%27%Male child (wrapped)²1956.357b30 yr
BC
to
AD
395(Roman) Thebes Resin on outer wrapping from areabetween calvesFat/oil
h
Coniferous resin90%9%Male child (unwrapped)²1956.357c30 yr
BC
to
AD
395(Roman) Thebes Resin from abdominal cavity (kidneyarea)Fat/oil
 j
Coniferous resinBalsam (?)87%13%0.1%
...................................................................................................................................................................................................................................................................................................................................................................
Several samples were taken from all but one mummy (unwrapped male child). For example, 15 different samples were analysed from various parts of the body of Horemkenesi.* Manchester Museum; ² National Museum of Scotland; ³ Liverpool Museum; § Bristol Museum;
k
British Museum.Museum number.
#
The term `resin' denotes physical appearance and does not presuppose any chemical composition or biological origin.
I
Superscript letters (a±w) refer to the histograms shown in Fig. 1.** Percentage relative abundance based on absolute concentrations, calculated on the basis of internal standards added at the extraction stage. Compositions do not imply that they were the originalformulations of the embalmers, owing to possible chemical changes over time.
 © 
 
2001 Macmillan Magazines Ltd
 
times
14±17
.Thisstudy involvesthe®rstsystematicchemical investigationof acollectionofprovenancedEgyptianmummies datingfromthemid-dynastic period (
c.
1,900yr
BC)
to the late Roman period (
AD
395),which encompasses the period at which the mummi®cation processwas at its peak (1,350±1,000yr
BC
). We have used diagnostic markercompounds that are present in the original `balms', are resistantto degradation and can be related to speci®c embalming agents.Gas chromatography with mass spectrometry (GC-MS) and ther-mal desorption (TD)- or pyrolysis (Py)-GC-MS facilitate themolecular separation and identi®cation of the marker compounds.Because of the probable nature, possible processing and burialhistory of embalming materials, both free and polymerized com-ponents are likely to be present. Therefore, this `dual' approachallows the characterization and identi®cation of both the free(solvent-extractable) marker compounds and the recognizablesubunits of polymeric materials not amenable to the more conven-tional GC-MS approach
17
. Given the understandable sensitivity surrounding the studyof mummies, the amount of sample requiredis an important consideration. Sequential TD-GC-MS and Py-GC-MS require very small sample sizes (
,
0.1mg), facilitating thevirtually non-destructive analysis of valuable museum specimens.A summary of the results of this study is given in Table 1. Themain products seen in all mummies are degraded acyl lipids, whichare derived from plant oils and animal fats (Fig. 1a±m). Wheresamples were taken directly from the bodies, these may well derivefrom endogenous body lipids, such as triacylglycerols from fats orcell-membrane phospholipids. But even after accounting for decay,for the most part these display non-human fatty-acid distributions.In cases where degraded fats or oils are, for example, seen in thebandaging and are not directly in contact with the body, they mustderive from their intentional application as part of the embalmingprocess.Inmostcases,thefatty-aciddistributions (Fig.1)areindicativeof plant origins; that is, they have a high abundance of C
16:0
comparedwith their abundance of C
18:0
. But it is not possible to assign aprecise plant origin on the basis of fatty acids alone, owing todegradation of the major unsaturated components; such degrada-tionisevidentfromthepresenceofmono-anddihydroxycarboxylicacids, which are indicative of autoxidation. The presence of plantoils (and to a lesser extent animal fats) suggests that they were key ingredients in mummi®cation, and were probably used as a less-costly base with which to mix and apply more exotic embalmingagents to the bodies and/or wrappings.Their widespread use indicates that the embalmers were aware of the special properties of unsaturated oils and fats that allow them to`dry', or rather, to polymerize spontaneously 
18
(later also appre-ciated by the oil painters of western Europe). This polymerizationwould have produced a highly crosslinked aliphatic network, whichwould have stabilized otherwise fragile tissues and/or textile
letters to nature
838
NATURE
|
VOL 413
|
25 OCTOBER 2001
|
www.nature.com
100 1001000
a
1000
b
1000
c
×
100
d
1000
e
1000
f
×
101000
161616161616161616161616162020202020202020202020202024242424242424242424242424282828282828282828282828283212121212121212121212121212
1000
l
×
101000
m
1000
 j
×
100
i
1000
h
1000
g
0100
312923 332527
n
0100
313131312929292923232323333333332525252527272727
o
0100
p
0100
q
0100
1000
5050505050484848484842424242424040404040525252525244444444444646464646
s
0100
t
0100
u
0100
 v 
0100
 w 
   R  e   l  a   t   i  v  e  a   b  u  n   d  a  n  c  e   (   %   )
Number of carbon atoms
Figure 1
Chemical analyses of samples taken from the mummies in this study.Histograms show the distribution of fatty acids of equal carbon number in acid fractions
a
±
m
; see Table 1), hydrocarbons of odd carbon number in neutral fractions fromsamplescontainingbeeswax
n
±
r
 ), waxestersof even carbonnumber inneutralfractionsfrom samples containing beeswax ( 
s
±
w
 ).
 © 
 
2001 Macmillan Magazines Ltd
 
wrappings against degradation by producing a physico-chemicalbarrier that impedes the activities of microorganisms.Theresultsfromthewidedaterangeofthemummiesinvestigatedreveal notable trends in the evolution of the use of other commod-ities (Fig. 2). For example, beeswax and coniferous resin clearly increase in their prominence through time, and are found inmaterial taken both directly from the bodies and from the wrap-pings. Coniferous resin is identi®ed by the presence of bothfunctionalized and defunctionalized diterpenoid components. Forexample, 7-oxodehydroabietic acid and 15-hydroxy-7-oxodehydro-abietic acid were usually the dominant diterpenoid components,and the normally abundant dehydroabietic acid was virtually absent.Although coniferous resins were clearly used in the embalmingprocess at least as early as 2,200yr
BC
(VI dynasty)
16
, their usebecomes most apparent in later periods; both the tissues and thewrappings of mummies from the Roman period (30yr
BC
to
AD
395)contain appreciable quantities (up to 37%) of coniferous diter-penoids. The increasing use of coniferous resin suggests that theembalmers may have become aware of the ability of speci®c naturalproducts to inhibit microbial degradation by means of mechanisms(physico-chemical barriers and antimicrobial action) analogous totheir protective roles in the plants from which they derived.A signi®cant trend is also seen in the use of beeswax, which ischaracterized chemically by alkanes (C
25
±C
33
) (Fig. 1n±r), wax esters (C
40
±C
50
) (Fig. 1s±w) and hydroxy wax esters (C
42
±C
54
). It®rst appears notably later than coniferous resin, with its positiveidenti®cation in a resinous coating taken from the chest cavity of afemale mummy of the Third Intermediate Period (XXI to XXVdynasty; 1,069±664yr
BC
). In a sample taken from `Pedeamun', aXXVIdynasty(664±525yr
BC
)mummy,thesolvent-soluble extractscomprise 38% beeswax (Fig. 3). But an even greater amount wasfound in a very extensive black `resinous' layer from a femalemummyofPtolemaicdate(332±30yr
BC
)(Fig.4),inwhichbeeswamade up 87% of the solvent extractable components; in otherwords, this treatment was essentially beeswax mixed with a smallamountof 
Pistacia
resin(seealsobelow).Thechoiceofbeeswaxwasclearly motivated by its hydrophobic/sealant and antibacterialproperties, notwithstanding its symbolic signi®cance
19
.Othercomponents of the embalming `resin' are triterpenoids andhydroxyaromatic acids, which are diagnostic of plant products. The
letters to nature
NATURE
|
VOL 413
|
25 OCTOBER 2001
|
www.nature.com
839
2500yr BC1500yr BC500yr BC500yr AD02000yr BC1000yr BCDate
   C  o  m  m  o   d   i   t  y
BeeswaxConiferresinPistacia/ triterpenoidresinPlant oilBitumenBalsamic (?)resin*1 1111 111,21,41 1111,21,4241113112114
Figure 2
The use of organic commodities (preservatives/unguents) identi®ed in chemicalinvestigations of the `balms' of provenanced and dated ancient Egyptian mummies. Greyscale re¯ects the relative abundance of a particular commodity found in the balmsamples, with darkershades representing a higher relative abundance. Asterisk, tentativeidenti®cation of balsamic resins; 1, data from this study; 2, data from ref. 14; 3, data fromref. 16; 4, data from ref. 15.
251051520
   C
   4   0
   C
   2   5
   C
   3   1
   C
   2   9
   C
   3   3
   C
   3   0
   C
   2   8
   C
   2   7
   C
   4   2
   C
   4   4
   C
   4   6
   C
   4   8
   C
   4   4
   O   H   C
   4   8
   O   H   C
   5   0
   O   H   C
   4   6
   O   H   C
   5   0
IS
   C
   1   6  :   0
   C
   1   6  :   0
        δ
   C
   1   6  :   0
      γ
   C
   1   8  :   0
Retention time (min)
   R  e   l  a   t   i  v  e   i  n   t  e  n  s   i   t  y   C
   1   6  :   0
   C
   1   7  :   0
   C
   1   5  :   0
   C
   1   4  :   0
   C
   2   0  :   0
   C
   2   2  :   0
   C
   2   4  :   0
   C
   2   6  :   0
   C
   2   8  :   0
   C
   3   0  :   0
   C
   1   8  :   0
IS
   C
   7
   C
   8
   C
   1   0
   C
   9
   C
   6
510 15 20
O
 
CO
2
HCO
2
H
Figure 3
Reconstructed total-ion chromatogram of the trimethylsilylated neutral fraction(after solvent extraction and fractionation) of `resin' from the top of the cranium of the Lateperiod male adult `Pedeamun', XXVI to XXVII dynasty (664±404 yr
BC
 ). Peak identities
indicates carbon chain length): ®lled triangles, C
:0
indicates saturated fatty acid methylesters; ®lled squares, C
:0
indicates saturated lactones (carbon chain length
is followedby the position of the lactone ring); ®lled circles, C
indicates alkanes; open circles, C
indicates alkanols; ®lled inverted triangles, C
indicates wax esters; open invertedtriangles, C
OH indicates hydroxy wax esters; IS, internal standard ( 
-tetratriacontane).Inset displays a reconstructed total-ion chromatogram of the trimethylsilylated acidfraction (after solvent extraction and fractionation) of this sample. Peak identities: opentriangles,C
:0
indicatessaturatedfattyacids;opensquares,C
indicates
a
,
v
-dicarboxylicacids; open diamond, vanillic acid. Also shown are the structures of two diterpenoidacids identi®ed: dehydroabietic acid and 7-oxodehydroabietic acid. IS, internal standard
-heneicosanoic acid).
 © 
 
2001 Macmillan Magazines Ltd

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