Biomolecular analyses enable new insights into ancient Egyptian embalming

Abstract

The ability of the ancient Egyptians to preserve the human body through embalming has not only fascinated people since antiquity, but also has always raised the question of how this outstanding chemical and ritual process was practically achieved. Here we integrate archaeological, philological and organic residue analyses, shedding new light on the practice and economy of embalming in ancient Egypt. We analysed the organic contents of 31 ceramic vessels recovered from a 26th Dynasty embalming workshop at Saqqara1,2. These vessels were labelled according to their content and/or use, enabling us to correlate organic substances with their Egyptian names and specific embalming practices. We identified specific mixtures of fragrant or antiseptic oils, tars and resins that were used to embalm the head and treat the wrappings using gas chromatography–mass spectrometry analyses. Our study of the Saqqara workshop extends interpretations from a micro-level analysis highlighting the socio-economic status of a tomb owner3–7 to macro-level interpretations of the society. The identification of non-local organic substances enables the reconstruction of trade networks that provided ancient Egyptian embalmers with the substances required for mummification. This extensive demand for foreign products promoted trade both within the Mediterranean8–10 (for example, Pistacia and conifer by-products) and with tropical forest regions (for example, dammar and elemi). Additionally, we show that at Saqqara, antiu and sefet—well known from ancient texts and usually translated as ‘myrrh’ or ‘incense’11–13 and ‘a sacred oil’13,14—refer to a coniferous oils-or-tars-based mixture and an unguent with plant additives, respectively.

Full text

Nature | Vol 614 | 9 February 2023 | 287
Article
Biomolecular analyses enable new insights
into ancient Egyptian embalming
Maxime Rageot1,2 ✉, Ramadan B. Hussein3,11, Susanne Beck3, Victoria Altmann-Wendling1,4,
Mohammed I. M. Ibrahim5, Mahmoud M. Bahgat5, Ahmed M. Yousef6, Katja Mittelstaedt1,
Jean-Jacques Filippi7, Stephen Buckley2,8, Cynthianne Spiteri2,9 &
Philipp W. Stockhammer1,10 ✉
The ability of the ancient Egyptians to preserve the human body through embalming
has not only fascinated people since antiquity, but alsohas always raised the question
of how this outstandingchemical and ritual process was practically achieved.Here we
integrate archaeological, philological and organic residue analyses, shedding new
light on the practice and economy of embalming in ancient Egypt. We analysed the
organic contents of 31 ceramic vessels recovered from a 26th Dynasty embalming
workshop at Saqqara1,2. These vessels were labelled according to their content and/or
use, enabling us to correlate organic substances with their Egyptian names and specic
embalming practices. We identied specic mixtures of fragrant or antiseptic oils,
tars and resins that were used to embalm the head and treat the wrappings using gas
chromatography–mass spectrometry analyses. Our study of the Saqqara workshop
extends interpretations from a micro-level analysis highlighting the socio-
economic status of a tomb owner3–7 to macro-level interpretations of the society.
The identication of non-local organic substances enables the reconstruction of trade
networks that provided ancient Egyptian embalmers with the substances required for
mummication. This extensive demand for foreign products promoted trade both
within the Mediterranean8–10 (for example, Pistacia and conifer by-products) and with
tropical forest regions (for example, dammar and elemi). Additionally, we show that at
Saqqara, antiu and sefet—well known from ancient texts and usually translated as
‘myrrh’ or ‘incense’11–13 and ‘a sacred oil’13,14—refer to a coniferous oils-or-tars-based
mixture and an unguent with plant additives, respectively.
Ancient Egyptians developed an outstanding ability to protect the
human body from decomposition or destruction after death—instigated
by the belief that the decomposition of the corpse presented a physi-
cal obstacle toward attaining the afterlife15. Performed by specialized
and learned individuals (ritualist embalmers), embalming was both
a chemical and a ritual process
14
. From a chemical perspective, the
practice evolved from simple natural preservation (through desic-
cation), via a proto-embalming treatment during prehistoric times
16
(around 4,300–3,100 ), to the sophisticated pharaonic procedures
of anthropogenic desiccation (using natron), excerebration, eviscera-
tion and the use of antibacterials, antifungals, barrier materials and
fixatives3,15. This preservation procedure, which could take up to 70 days
to complete, ensured the transformation of a vulnerable body into a
durable mummy. Embalming also entailed sets of ritualized acts and
the recitation of liturgical texts, through which the chemically treated
body would be revived and acquired a new identity as a justified or
glorified deceased, worthy of living on in the netherworld17.
Our present-day knowledge of embalming substances is derived
from two main sources: ancient written sources such as embalming
papyri14,18, and organic residue analyses (ORA) of Egyptian mummies.
Substances used in embalming have been named in ancient Egyp-
tian texts and by Greek authors such as Herodotus and Diodorus.
However, debates have arisen concerning the specific substancesto
which these terms correspond11,15,19,20. In recent years, ORA has been
applied to study residues recovered from mummies and embalming
vessels in individual tombs (for example, in ref.
3
). Although these analy-
ses have successfully identified various substances used in embalming,
the roles of these balms in this process as well as the overall procedure
have so far remained unclear.
The discovery of embalming facilities at Saqqara presented here
reshapes our knowledge and understanding of ancient Egyptian mum-
mification. Dated to around 664–525  (26th Dynasty), the embalm-
ing workshop is located a few metres to the south of the pyramid of
King Unas. It includes a subterranean evisceration facility (the wabet),
https://doi.org/10.1038/s41586-022-05663-4
Received: 10 August 2022
Accepted: 15 December 2022
Published online: 1 February 2023
Open access
Check for updates
1Institute for Pre- and Protohistoric Archaeology and Archaeology of the Roman Provinces, Ludwig Maximilian University of Munich, Munich, Germany. 2Department of Pre- and Protohistory,
Eberhard Karls University of Tübingen, Tübingen, Germany. 3Department of Egyptology, Eberhard Karls University of Tübingen, Tübingen, Germany. 4Department of Egyptology,
Julius-Maximilians University, Würzburg, Würzburg, Germany. 5The Central Laboratories Network, the National Research Centre, Cairo, Egypt. 6Packaging Materials Department, the National
Research Centre, Cairo, Egypt. 7Analytical Research Department, Robertet S.A., Grasse, France. 8BioArCh, University of York, York, UK. 9Department of Life Sciences, University of Turin, Turin,
Italy. 10Max Planck Institute for Evolutionary Anthropology, Leipzig, Germany. 11Deceased: Ramadan B. Hussein. ✉e-mail: maxime.rageot@uni-tuebingen.de; philipp.stockhammer@lmu.de
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288 | Nature | Vol 614 | 9 February 2023
Article
a multifunctional aboveground structure (probably corresponding to
the ibu) and communal burial spaces
1,2
(Fig.1; for a detailed descrip-
tion of the archaeological evidence, see Supplementary Information,
section1). In addition to these structures, a cache of embalming pottery
vessels was uncovered in the wabet facility. This cache includes a large
corpus of potsherds and both broken and complete vessels, with some
showing traces of burning as well as drippings of boiled substances
on their outer surfaces. Among the finds are 121 beakers and bowls
(a total of 59 ‘marl clay beakers’ and 62 ‘red goldfish bowls’; for shapes,
refer to Fig.1) inscribed with Hieratic and Demotic texts providing
embalming instructions (for example, ‘to put on his head’ or ‘bandage or
embalm with it’) and/or names of embalming substances (for example,
‘sefet’ or ‘dry antiu’) and sometimes with the title of an administrator
of the embalming workshop or the necropolis (Extended Data Table1).
Out of this corpus, we selected 9 beakers and 22 red bowls with the
most clearly readable labels for ORA. To establish a possible link with
the vessels from the wabet facility, we included in our analyses four
samples from two burial chambers (locations 3 and 4) at the bottom
of the communal burial shaft: two red bowls, one faience cup and one
red cylindrical vessel.
Organic residue analyses
A wide range of products was identified, including plant oils and tars,
resins, and animal fats (details in Extended Data Table1, Extended Data
Fig.1 and Supplementary Information, section 2).
Among the group of conifer by-products, juniper or cypress (here-
after juniper/cypress)by-products in the form of essential or fragrant
oil or tar were identified in 21 vessels (60%). Their identification is sup-
ported by the association of totarol derivatives and cuparene-related
sesquiterpenes21,22 (Fig.2). The cedar oil or tar is the second most
commonly detected product in the Saqqara vessels (19 vessels (54%)).
Its presence is indicated by the predominance or the equivalence of low
molecular weight sesquiterpenoids of the himachalene series over the
characteristic diterpenes of the abietane family22–24 (Fig.2).
With regard to angiosperm resins, we identified elemi in at least 15
vessels (43%) (Extended Data Fig.1) on the basis of the combination
of lupeol and α- and β-amyrin derivatives (Fig.3). This assemblage is
commonly associated with resin of Burseraceae, particularly that of
Canarium25–29 (also known as elemi) (Extended Data Table2). Bursera
and Protium resins could be excluded as they occur mainly in Central
and South America
21,30
. α- and β-11-keto amyrins were identified in the
15 vessels (sometimes together with their acetate derivatives), in two
cases together with traces of brein (urs-12-ene-3,16-diol). These bio-
markers are documented in elemis from the Asian rainforest25,26,29 but
elemis from the African rainforest should not be excluded (Extended
Data Table2). Finally, the compounds, olean-9(11),12-dien-3-ol
and urs-9(11),12-dien-3-ol were detected in 14 samples (Fig.3 and
Extended Data Table2). These were previously identified in artificially
aged elemis from Manila and Mexican copal
28
. In addition, Pistacia resin
was detected in five vessels (14%). The identification was on the basis of
the presence of characteristic biomarkers5,8,31 (for example, moronic,
oleanonic, isomasticadienonic and masticadienonic acids) (Fig.3).
Triterpenic markers of heat treatment8 were also identified in four
Burial chambers
ShaftK24
(30mdepth)
The ibu structureof the
embalming workshop
Embalmingroom(wabet)
ShaftM23-II
(13mdepth)
3
4
5cm
Fig. 1 | The e mbalming fa cilities a nd burial ch ambers of th e Saqqara
complex. Orange arrows show t he location s of the investig ated vessels . The
background image is a digital documentation of the Saqqara complex (copyright
M. Lang , Universität B onn).The two labelle d vessels were un covered in the
embalming ro om and corresp ond toa ‘red goldfis h bowl’ (inscri ption: ‘sefet +
dry antiu’) anda ‘whi teclay beaker’ (ins cription: ‘t o be put on his hea d’).
The unlab elledred bowlwith bla ck surface re sidue was uncovere d in the burial
chamber lo c. 4.
Compound identication AIDB-5
a: Cuparene 1,460
b: α-Dehydro-ar-himachalene 1,470
c: γ-Dehydro-ar-himachalene 1,489
d: ar-Himachalene 1,494
e: Longifolone 1,546
f: α-Muurolol (δ-cadinol), TMS 1,592
g: Nor-ar-himachalone 1,617
h: Himachalol, TMS 1,638
i: Oxo-ar-himachalol, TMS 1,651
j: Oxo-cuparene 1,658
k: Oxo-γ-dehydro-ar-himachalene 1,678
l: Farnesol or cedrol deriv, TMS
(tent.)
1,728
m: Oxo-hydroxy sesquiterpenoid,
TMS
1,969
n: Cuparenic acid, TMS 1,979
o: Retene 2,162
p: Methyl ether of totarol 2,186
q: Totarol, TMS 2,240
u: Dehydroabietic acid, TMS 2,329
v: Oxo-totarol, TMS (totarolone) 2,452
x: 7-Oxo-dehydroabietic acid,
TMS
2,529
10 14 18 22 26 30 34
SSTP017.
o
abcdeghk
l
m
n
C10:0
C18:0
C20:0
C22:0
MAG16
C18:1
C18:1
C17:0
Branched-C17:0
C16:0
C15:0
C14:0
C12:0
ij
f
SSTP024
Sefet
Dry antiu
p
quvx
Branched-C15:0
60 140 220
73
145 289
115
175
234
304
131
m/z
73
161
95 204
119
143 189
279
105
m/z
132
105 202
159 187
145
189
91
m/z
[M•+]
[M•+]
[M•+ -Me]
157
185
128
200
143
m/z
[M•+]
185
157
143
200
171
128
115
105
91
m/z
[M•+]
60 140 220
187
145
202
131
159
[M•+]
d
f
n
ab
c
219
l
ae
C12:0
C14:0
m
nC16:0
C17:0
C18:0
q
MAG16
A
(%)
tr (min)
TMSO
O
OTMS
Fig. 2 | Par tial gas chrom atograms o f organic res idue extract s from bowls
labelle d ‘dry antiu’ and ‘sefet’. Total ion chromato grams showing t he
molecular co nstituent s of the essen tial oil or tar of ced ar (brown) and junip er/
cypress ( purple), and animal fat (b lue). Sesquiterp enes andditerp enes are
labelled a–z. The p refix SST P is an identif ier for samples f rom the Saqqara Sa ite
Tombs Project . Right, elec tron ionizatio n mass spect ra (70 eV) of
characteristic corresponding sesquiterpenes from coniferous oils or tars.
MAG, monoacylglycerol. A, abundance;AIDB-5, ar ithmetic ret ention index; te nt.,
tentati ve assignment ;TMS, trimethyl silyl derivati ve; tr, retention time.
Content courtesy of Springer Nature, terms of use apply. Rights reserved

Nature | Vol 614 | 9 February 2023 | 289
100
100
23
27
28
47
21
50
52
51
55
59
24
25
1016
78
62 29
39 40
57
5
3411 18 32
34
33
37
38
54
Modern resin of
Canarium strictum
30-year-old resin of
Canarium luzonicum
56
47.5 48.0
64 66
67
42 43
SSTP011
23 27
28
50
52
55
3
4
7811
21
23
27
28 32
33
47 52
50 55
100
38
0
30-year-old resin of
P. lentiscus (mastic)
1
931
8
11
14
17 19
21
23
41
45
49
51 58
63
61
26
13
22
27 30
43
44
15
tr (min)
Previous content
12
1
1
9
9
11
11 31
46
51
51
51
21
19
17
17
14
14
23 45 58 63
W40
W42 W44 W46
W48
41 42 43 44 45 46
A (%)
Last deposit
SSTP001a
SSTP001b
0
40′46′56 6564
46
a
100
100
100
100
30-year-old resin of
Shorea selamica (dammar)
20 26
36 42 48 56 62
35
1
9
11
12
15
22
27 30
31
43 46
51′
46′
40′
Surface residue
Absorbed residue
60 c
218
408 664.7
189
203
232
135
407
273
422
175
678.7
422
407
273
232
135
175
678.7
m/z
m/z
m/z
[M•+]
[M•+]
[M•+]
64
66
67
100 200 300 400 500600
A (%)
b
t
r
(min)
OO
O
OO
O
O
O
Bandage/
wrap/
embalm
with it
Fig. 3 | Par tial gas chr omatogra ms showin g the molecu lar const ituents of
Pistacia resin, dammar and beeswax. a,b, Total ion chromato grams showi ng the
molecular constituents of Pistacia resin (visible surface residue), dammar and
beeswa x (absorbed re sidue) (a) and elemi (b). c, Electron ioniz ation mas s spectra
(70eV) of triterp enic palmit ates. Gre en circles indi cate markers of Pistacia resin,
pink circles i ndicate marker s of dammar res in and yellow circle s indicate marke rs of
elemi. Fille d circles are bio markers prese nt in fresh re sin, empty ci rcles are
degrada tion markers linke d to natural oxida tion and/or hea t treatmen t and
half-filled cir cles are biomar kers and/or degr adation marker s. Number s prefixed
with W are the nu mber of carb on atoms in the l ong-chain es ters asso ciated with
the corres ponding pea k. Triterpenes a re labelled num erically as follow s: 1,
28-norolean-12, 17-dien-3-one; 2, olean-9(11),12-dien-3-one; 3, 3-epi-β-amyrin; 4,
3-epi-α-amy rin; 5, 3-epi-lup eol; 6, olean-9(1 1)-en-3-one; 7, urs-9(11),12- dien-3-one; 8,
olean-9(11),12-dien-3-ol; 9, nor-β-amyrone (28-norolean-12-en-3-one); 10,
α-amyrenone isomer (urs-9(11)-en-3-one); 11, β-amyrenone; 12, dammaradien-3-one; 13,
olean-18-en-3-one; 14, 28-noroleandien-3-one or28-norursdien-3-one(tent.); 15,
nor-α-amyrenone (28-norurs-12-en-3-one); 16, urs-9(11),12-dien-3-ol; 17,
28-norolean-17-en-3-one; 18, olean-3,12-dien-16-ol (dehydroxymaniladiol); 19,
oleandienol; 20, nor-β-amyrin (28-norolean-12-ene-3-ol); 21, α-amyrenone; 22,
dammaradien-3-ol (3β-hydroxy-20,24-dammarediene); 23, β-amyrin; 24, lupenone;
25, olean-9 (11),12-dien-3-yl a cetate; 26, n or-α-amyrin (28-noru rs-12-ene-3-o l); 27,
α-amyrin; 28, lup eol; 29, ursa-9(11),12-di en-3-yl acetat e; 30, δ-amyrenon e
(olean-13(18)-en-3-one); 31, noroleanenol ornorursenol(tent.); 32, maniladiol
(olean-12-ene -3,16-diol); 33, 11- oxo-β-amyrin ep i-isomer (tent.); 3 4, 11-oxo-α-amyrin
epi-isome r (tent.); 35, dammare nolic acid; 36, sho reic acid; 37, lupeol isome r; 38,
brein (urs-12-ene -3,16-diol); 39, β-amyrin ace tate; 40, α-amyrin a cetate; 40 ,
20,24 -epox y-25-hydrox ydammar en-3-on e; 41, hydroxyd ammara dienon e (tent.);
42, oleandien-28-ol (tent. erythro-3-en-28-ol); 43, hydroxydammarenone; 44,
oleanonic a ldehyde; 45, moronic a cid; 46, oleano nic acid; 46, 20,24-ep oxy-
25-hyd roxydamm aren-3- ol; 47, 11-oxo-β-amy renone; 48, hydrox ydammar enol; 49,
oleanol der ivative orursol d erivative; 50, 11- oxo-α-amyrenone; 5 1, oleanolic a cid;
51,ursonic acid (3-oxours-1 2-en-28-oic acid );52, 11-oxo-β-amyri n; 53, ursolic
aldehyde; 54, ole anolic aldehyde; 5 5, 11-oxo-α-amyrin; 5 6, ursolic aci d; 57, lupane
derivati ve (tent. canari c acid); 58, isoma sticadie nonic acid; 59, 11-oxo-β-amy rin
acetate; 6 0, 11-oxo-α-amyrin ac etate; 61, 11- oxo-oleanoni c acid; 62, hydroxy
oleanolic a cid; 63, mastic adienonic a cid; 64, α-amyrin palm itate (urs-12-en-3-yl
palmitate); 65, oxo-oleanene palmitate; 66, 11-oxo-β-amyrin palmitate; 67,
11-oxo-α-amyrin palmitate.
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290 | Nature | Vol 614 | 9 February 2023
Article
vessels (Extended Data Fig.1). A Dipterocarpaceae resin, commonly
known as dammar, was detected in one red bowl from burial chamber,
location 4. This resin is characterized by a broad assemblage of triter-
penic markers from dammarane, nor-ursane and oleanane families.
Although some of these biomarkers are ubiquitous, the co-occurrence
of dammaradien-3-ol, nor-α-amyrone, δ-amyrone and oxidation prod-
ucts such as 20,24-epoxy-25-hydroxydammaren-3-ol is a convincing
argument for the identification of dammar. To our knowledge, these
compounds have not been found together in any other resin31–34 (Fig.3).
Animal fat was detected in 18 vessels (51% of vessels). Its presence was
indicated by a narrow distribution of saturated triacylglycerols (TAGs)
(46:0 to 54:0 (carbon atoms:unsaturated carbon–carbon bonds)) and
diacylglycerols
35
(32:0 to 36:0). Furthermore, traces of saturated TAGs
with an odd number of carbon atoms (53:0, 51:0 and 49:0), which are
characteristic of ruminant animal fats
36
, were identified in 7 vessels
(Extended Data Figs.1 and 2). Plant oils were detected in 5 vessels (14%).
In 4 of them, a plant oil, type olive (although degraded argan or hazel-
nut oils cannot be excluded) was indicated by the specific distribu-
tion of unsaturated TAGs (54:3, 52:2, 50:1) and diacylglycerols37 (34:1,
36:2 and 32:0) (Extended Data Fig.3). The detection of ricinoleic acid
together with a substantialamount ofoleic acid and its mono- and
dihydroxylated degraded markers in one beaker suggests that it may
have contained castor oil, possibly mixed with other oils
38–40
(Extended
Data Fig.4). Although ricinoleic acid has also been associated with
the activity of ergot fungi on Gramineae
41
, the castor oil hypothesis
remains the most plausible in the Saqqara context, where embalming
vessels were dedicated to the preparation of antiseptic and antifungal
substances for mummification. Beeswax was identified in 5 vessels
(14%) by the presence of its characteristic even-numbered fatty acids
(22:0 to 28:0, with 24:0 being the most important) and long-chain
(C40 to C48) palmitic esters5,42 (Fig.3).
Bitumen was found in two vessels recovered from the burial cham-
bers at locations 3 and 4, based on the characteristic hopanes and
steranes
3,23,43,44
(Extended Data Fig.5). Its chemical composition sug
-
gests that it originated from the Dead Sea23 (Supplementary Informa-
tion, section2).
Finally, we identified molecular markers of recipes that involve the
mixing and heating of resinous substances with fat or oil in three vessels
(triterpenic palmitates; Supplementary information2 and ref.
45
). Elemi
was prepared together with animal fat and/or plant oil in two beakers
and dammar was prepared with beeswax and/or animal fat in a bowl
(Fig.3 and Extended Data Fig.1).
Treatment of the body in the workshop
The inscriptions on the vessels of the Saqqara workshop contain
instructions for the treatment of specific body parts, especially the
head, and for the preparation of linen bandages. Some of these treat-
ments involved the preparation and application of several mixtures.
Eight vessels are inscribed with instructions for the treatment of the
head. Our samples show that the embalmers used three different mix-
tures (mixtures A, B and C in Fig.4 and Extended Data Fig.1), which can
include elemi, Pistacia resin an oil or tar of juniper/cypress and cedar,
animal fat, beeswax, probably castor oil, and a plant oil (type olive).
To our knowledge, the use of elemi and oil or tar of juniper/cypress
for embalming the head has not previouslybeen reported. However,
previous ORA studies of early mummies from the first millennium
suggest, in accordance with our results, that castor oil and Pistacia
resin were used specifically for the treatment of the head
6,40,46
. Beeswax,
Pinaceae by-product, and fat or oil were additionally used for different
parts of the body3–6.
We extracted samples from eight vessels (four beakers and four red
bowls) with labels indicating for ‘wrapping or embalming with it’, which
were probably used for preparing mummy linen bandages. The organic
contents of seven vessels were mixtures (mixtures D and E; Fig.4 and
Extended Data Fig.1), and one bowl contained only animal fat. Mixture E
was the most frequently detected (five vessels) and consisted of oil or
tar of juniper/cypress and cedar, animal fat and/or plant oil and elemi.
In two of these vessels, we additionally identified heating markers of
elemi resin together with fat or oil. Previous studies of mummy band-
ages from the 4th millennium  and later provide evidence for the use
of fat or oil and conifer by-product in most of the balms, but none for the
use of elemi
3–6,16
. However, one sample from the 1st millennium  was
treated with a mixture including fat or oil, a conifer by-product and a
triterpenic resin resembling mixture E
5
. Previous studies have provided
evidence that bitumen and beeswax were regularly incorporated into
Mix A
Elemi, Pistacia resin, oil or tar
of juniper/cypress and cedar
(can be augmented with castor oil,
animal fat and/or beeswax)
Mix D
Oil or tar of juniper/cypress and/or
elemi, animal fat and/or plant oil
Mix E
Oil or tar of juniper/cypress
and cedar, animal fat and/or plant
oil, elemi (sometimes heated)
“To wash”
Oil or tar of conifer
"To make his odour pleasant"
Ruminant animal fat (adipose or
dairy), degraded Burseraceae resin
Oil or tar of juniper/cypress
and elemi
Heated beeswax
Treatment of bandage, wrap or embalming
Treatment of the head
Imseti (protects liver)
Mix B
Elemi, oil or tar of cedar,
(plant oil type olive)
Mix C
Elemi, animal fat, plant
oil type olive
Skin treatment (3rd day of treatment)
Ruminant animal fat (adipose or
dairy), heated beeswax
Animal fat
Duamutef (protects stomach) []
Fig. 4 | Organic contents of vessels providing embalming instructions. Organi c substance s and/or mixtures i dentifie d in the potter y and the inscr iptions
associa ted with thes e vessels. Mu mmy drawing copyr ight S. Luca s.
Content courtesy of Springer Nature, terms of use apply. Rights reserved

Nature | Vol 614 | 9 February 2023 | 291
balms for bandages during this period
3–5,7
. However, neither of these
substances were detected in vessels used for the mummy bandages at
Saqqara (although the limited amount of residue absorbed prevented
the application of targeted methods6,47). Instead, we found two new
substances—elemi and juniper/cypress.
Six other sherds provided information on substances used for wash-
ing the body, reducing bodily odour and softening the skin, as well as
a recipe for the treatment of the liver and another for the stomach.
The bowl labelled with ‘to wash’, contained markers of oil or tar of
conifer, and the bowl inscribed with ‘to make his odour pleasant’
showed evidence of ruminant animal fat (adipose or dairy) and
degraded Burseraceae resin (Extended Data Table2). In the vessel
with inscriptions related to the treatment of the skin, which may have
occurred on the third day of embalming (Extended Data Table1), we
identified a mixture of animal ruminant fat (adipose or dairy) com-
bined with heated beeswax.
Two of the sampled vessels were inscribed: one with the name of the
god Imseti, who protects the liver, and the other with the god Duamutef,
who protects the stomach. One of these vessels (Imseti/liver) contained
a mixture of oil or tar of juniper/cypress and elemi, whereas the other
(Duamutef/stomach) contained only heated beeswax (potentially
similar content of two 26th Dynasty canopic jars is described in ref. 7).
Another bowl was inscribed with the title of an administrator of the
embalming workshop and the necropolis—the seal bearer—who carried
out specific embalming procedures, related mainly to the treatment of
the head
14
. This vessel yielded fat or oil and oil or tar of juniper/cypress,
which is identical to mixture D, for treating linen bandages and which
could have been used to wrap the head.
Embalming vessels in the burial chambers
The embalmers of the workshop also provided additional services,
including the burial of the deceased in communal burial spaces
1
. We
analysed four vessels from two communal burial chambers (locations
3 and 4) to evaluate similarities and differences among the substances
used during burial.
One bowl from location 4 was used multiple times and for different
substances. A visible black residue lining its surface was identified as a
pure heated Pistacia resin. However, the ceramic sample taken from its
inner wall showed markers of oils or tars of cedar and juniper/cypress,
bitumen and dammar mixed with beeswax and/or animal fat. This points
to the complex and extended usage of the vessel, used first to prepare
the different substances (ceramic impregnation) and subsequently to
contain a heated Pistacia resin (last deposit).
From burial chamber location 3, we analysed a small faience cup
and a red cylindrical pottery vessel. The cup still contained a cake-like
substance, consisting of oil or tar of cedar, animal fat, heated Pistacia
resin and heated beeswax. The cylindrical vessel contained oil or tar of
cedar and possibly of juniper/cypress as well as bitumen and a fat or oil.
With the exception of the dammar and bitumen, all the substances
detected in the vessels recovered from the burial chambers matched
those identified in the embalming workshop.
Properties and management of substances
These results suggest that the embalmers used the substances for their
specific biochemical properties, as Pistacia resin, elemi, dammar, oils,
bitumen and beeswax have antibacterial or antifungal and odorifer-
ous properties, and thus help to preserve human tissue and reduce
unpleasant smells
4,33,42,44
. Animal fat, plant oil and beeswax were also
essential ingredients in recipes for the treatment of different body
parts, as well as in ointments used to moisturize the skin48. Finally,
the hydrophobic and adhesive properties of tars, resins, bitumen and
beeswax were useful to seal skin pores, exclude moisture and to treat
linen wrappings. The colour or appearance of these products may also
have been desirable4.
The embalming substances identified point to the existence of a
management system of bio-products, from harvest, transportation,
Saqqara
Saqqara
Saqqara Saqqara
Saqqara
Pistacia resin
Elemi
Dammar
Cedar
Juniper or cypress
Bitumen
Fig. 5 | Poten tial origi ns of impor ted bioreso urces at Saq qara complex. C oloured areas in dicates the po tential orig ins of the raw mater ials that were use d for
the preparat ion of balms and th e mummifica tion process es at Saqqara . Map copyright S . Lucas.
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292 | Nature | Vol 614 | 9 February 2023
Article
transformation and application. For example, obtaining plant oil and
animal fat necessitate an extraction system, and the production of
wood tar (pyrolysis) or oil (steam distillation) involves thermal process-
ing and the specific controlled management of the raw material
49
. In
addition, the thermal treatment of substances (such as Pistacia resin
and beeswax) and the subsequent production of recipes (for exam-
ple, those based on elemi and dammar resins) required specialized
knowledge, technical skills and tools to obtain balms with the desired
properties. Our results demonstrate that the embalmers indeed car-
ried out activities that require specific know-how and benefited from
institutional organization.
Antiu and sefet
An important challenge for understanding Egyptian embalming prac-
tices on the basis of textual sources has always been the translation of
substance-related terms
20
. Lexicographically, antiu has tentatively
been associated with myrrh on the basis of philological conjectures
11–13
.
However, five vessels from the embalmers’ workshop that carry the
label antiu yield a mixture of oil or tar of cedar and juniper/cypress
together with animal fat (Extended Data Fig.1; the use of cedar and/
or juniper/cypress oil in ancient Egypt is described in refs.
22,23,46,50,51
).
The labels indicate that antiu could have been used alone in dry form
or mixed with sefet. However, in all cases we find markers of a mixture
of coniferous volatile products with animal fat. This strongly suggests
that antiu is a product that was purposefully manufactured by the
embalmers and whose preparation entails the transformation of at least
two different coniferous oils or tars and then mixing them with animal
fat. In the Saqqara context, translations of antiu as a raw material such
as myrrh can be excluded.
In Egyptology, sefet is usually described as an unidentified oil12,13,48.
It was one of the ‘7 sacred oils’ that were used in embalming and the
‘opening of the mouth’ ritual
13,14
. In three vessels from the embalm-
ers’ workshop with the label ‘sefet’, we identified markers of animal
fats, which were mixed in two of these vessels with oil or tar of juni-
per/cypress. The third vessel contained the markers of ruminant fat
(adipose or dairy) with elemi. This indicates that, at least at Saqqara,
sefet was a scented unguent (fat-based formula) with plant additives,
particularly Cupressaceae or Burseraceae by-products. It is possible
that the scented sefet unguent was also prepared with other plant oils.
Moreover, its composition may have evolved over time14,46,51.
Egyptian mummification and the world economy
The majority of the substances used at the Saqqara workshop were
imported—many of them from a considerable distance. The Saqqara
context (Extended Data Figs.6, 7 and 8) provides only a glimpse into
the trade and exchange systems required to run a comprehensive
embalming industry
3,15,52
. These findings confirm the known pattern
of the diversification and complexification of embalming practices
after around 1000 3,5. The origin of the different substances provides
evidence for an almost global network (Fig.5). The bitumen identified
in Saqqara most probably originated from the Dead Sea, confirming
previous findings that the asphalt from this region was exported to
Egypt in the first millennium  specifically for mummification
4,53
.
Pistacia trees producing high yields of resin (Pistacia lentiscus or
Pistacia terebinthus), olive trees, cedar, juniper and cypress are absent
in Egypt
8,11,21,30
, but grow in different locations in the Mediterranean
basin (Fig.5). The related by-products were also imported, most prob-
ably from the Levant (for example, Cedrus libani), which had important
trade networks with Egypt8–10.
Although intensified trade networks and cross-cultural exchanges
are well-documented for the regions of the Mediterranean basin, the
Saqqara workshop provides additional evidence for long-distance trade
networks via the vivid Indo-Mediterranean trade routes, which seem
to have existed since the 2nd millennium 54. This is particularly true
for resins, which are endemic to rainforests. Canarium species, which
produce elemi, are distributed in both Asian and African rainforests
21,30
,
whereas dammars are harvested from Dipterocarpaceae trees that
grow exclusively in Asian tropical forests
21,30
. Thus, it is possible that
elemi reached Egypt by the same route as dammar
55
. Consequently,
the embalming and funerary services of the 7th century  Saqqara
workshop kept the demand for such biomaterials from distant lands
active and supported the flourishing of international trade networks
connecting Egypt with the eastern Mediterranean in addition to Asian
and possibly African rainforests.
Conclusion
We have identified several specific mixtures used for embalming the
head or wrapping the body. The mummification specialists seem to
have been aware of both the chemical properties and the bioactivity of
the substances used and to have obtained complex knowledge about
the preparation of different balms of particular ingredients. We identi-
fied antiu and sefet as mixtures of different fragrant oils or tars and fats.
Antiu should be less restrictively designated—that is, not exclusively as
myrrh or incense. Egyptian mummification was built upon and fostered
long-distance exchange and routes, including imports from the Medi-
terranean basin as well as Asian and possibly African rainforest regions.
Online content
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ries, source data, extended data, supplementary information, acknowl-
edgements, peer review information; details of author contributions
and competing interests; and statements of data and code availability
are available at https://doi.org/10.1038/s41586-022-05663-4.
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Article
Methods
Upon the discovery of the embalming vessels of the Saqqara workshop,
a multinational team of researchers from the Universities of Tübingen
and the Ludwig Maximilian University of Munich (Germany), and the
National Research Centre (NRC) of Cairo (Egypt) was formed. Vessels
were sampled on site at Saqqara and samples were delivered to the
NRC laboratories for extraction and analyses.
Sample treatment before GC–MS analyses
ORA was carried out at the NRC, Chromatographic Laboratories
Network, Giza, Egypt. One gram of pottery powder was drilled out
from the inner walls of the vessel (layer 2), following cleaning of its sur-
faces in order to remove any exogenous lipids. The characterization
of the lipid constituents present was based on the analytical results
obtained from layer 2. The ceramic powder collected during sur-
face cleaning (layer 1) was retained for potential additional analysis.
Powdered sherds were solvent-extracted (dichloromethane:methanol,
2:1 by volume) by ultrasonication to target lipid and resin compounds
following established protocols56. 50% of the total lipid extract were
trimethylsilylated (40°C for 20min) using N,O-bis(trimethylsilyl)
trifluoroacetamide (BSTFA) (50µl) and a catalytic reagent (pyridine)
(4µl) before analysis by gas chromatography–mass spectrometry
(GC–MS).
Modern and aged (30-year-old) angiosperm resins, which included
Pistacia, dammars, frankincense, elemi and myrrh (Extended Data
Table3) were ground, then extracted by ultrasonication in dichlo-
romethane (1mgml
−1
) and trimethylsilylated following established
protocols49.
Gas chromatography and GC–MS analyses
The analysis of trimethylsilylated samples was performed by GC–MS
using an Agilent 7890B GC system and Agilent 5977 MSD.
The analyses were carried out using helium as a carrier gas, with a
split/splitless injection system (SSL), operating in the splitless mode
with a flow rate of 3.0mlmin
–1
of helium and a constant pressure at
the head of the column of 8.6667psi. Samples were analysed using an
Agilent J&W DB-5HT-column (15m × 0.32mm internal diameter; 0.1µm
film thickness). The temperature of the oven was set at 50°C for 1min
then ramped to 100°C at 15°C min
–1
, then to 240°C at 4°C min
–1
and
to 380°C at 20°C min
–1
(held isothermally for 7min). The inlet tem-
perature was set at 300°C. Mass spectra were acquired using electron
ionization at 70eV and obtained by scanning between m/z values 50
and 950. The interface and the ion source temperatures were 300°C
and 230°C, respectively.
Some samples composed of triterpenoid markers and determined to
be free of high molecular weight components (absence of wax esters,
TAGs, triterpene palmitate) by the conditions described above, were
analysed using an Agilent J&W DB-5MS column (30m × 0.25mm internal
diameter; 0.25µm film thickness). The inlet temperature was fixed at
300°C. The oven temperature was ramped from 50°C (held isother-
mally for 1min) to 150°C at 10°C min
–1
, and then increased to 320°C
at 4°C min–1 (held isothermally for 15min). The analyses were carried
out using helium as a carrier gas, with a flow rate at 2.0mlmin
–1
and the
operating in the splitless mode with a purge flow of 3.0mlmin–1 and a
split ratio of 3:1. Mass spectra were acquired using electron ionization
at 70eV. The mass range was scanned for m/z 50–950. The ion source
temperature was set at 230°C and the transfer line at 250°C.
Chromatograms and mass spectra were matched against authentic
standards (lupeol, lupenone, α- and β-amyrin, saturated and unsatu-
rated triglycerides, fatty acids, n-alkanes)8,22,28,31,57–59 and the National
Institute of Standards and Technology (NIST) library60.
Retention indices were calculated based on a series of straight chain
hydrocarbons from 7 to 40 carbons and were also used to confirm
the identification of sesquiterpenes and diterpenes. The arithmetic
retention indexes (AI) used in ref. 59 were computed as: AI(x)=100z+
100[(RT(x)−RT(Pz))/(RT(Pz+1)−RT(Pz))], according to Van den Dool
and Kratz61;x, analyte; RT, retention time; Pz are paraffins (n-alkanes)
with z carbon atoms.
Reporting summary
Further information on research design is available in theNature Port-
folio Reporting Summary linked to this article.
Data availability
All information on the samples and the data generated and analysed in
this study is included in the manuscript, supplementary information
files and Extended Data files.
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chromatographic, mass spectrometric and stable isotope approaches to the classiication
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Acknowledgements This study was funded by the European Research Council (ERC)
under the European Union’s Horizon 2020 research innovation programme (ERC-2015-
StG 678901-Food-Transforms) as part of ERC Starting Grant project ‘FoodTransforms:
Transformations of Food in the Eastern Mediterranean Late Bronze Age’ (P.W.S.) and the DFG
Project Saqqara Saite Tombs Project (directed by R.B.H.; project number 288139336). The
authors thank M. Regert for providing some reference resins and K. Ryholt for helping with the
Demotic inscriptions.
Author contributions M.R. and R.B.H. designed and performed the research. M.R. and A.M.Y.
sampled the pottery at Saqqara. M.R., K.M. and M.I.M.I. processed the ORA of ceramic sherds.
M.R. interpreted the organic residue data with the help of J.-J.F. and S. Buckley. R.B.H. directed
the excavation of the site of Saqqara. R.B.H., S. Beck and V.A.-W. performed the study of
Hieratic and Demotic texts and provided historical background. M.R., R.B.H. and P.W.S. wrote
the paper. M.M.B. and C.S. proofread the manuscript.
Competing interests The authors declare no competing interests.
Additional information
Supplementary information The online version contains supplementary material available at
https://doi.org/10.1038/s41586-022-05663-4.
Correspondence and requests for materials should be addressed to Maxime Rageot or
Philipp W. Stockhammer.
Peer review information Nature thanks Carl Heron and the other, anonymous, reviewer(s) for
their contribution to the peer review of this work.
Reprints and permissions information is available at http://www.nature.com/reprints.
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Extende d Data Fig. 1 | Or ganic sub stances i dentif ied in the pot tery fro m the Saqqara e mbalming wor kshop and co mmunal buri al spaces . Flame = molecular
markers ass ociated wit h heat treatm ent of the subst ance. Dot ted squares = m arkers present i n the substan ces, but the as semblage is not s pecific e nough.
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Article
Extended Data Fig. 2 | Characteristic biomarkers of ruminant fat. Partial Total Ion Ch romatograms a nd mass spec tra (EI, 70eV) showing even a nd odd
saturated triacylglycerols (TAGs).
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Extended Data Fig. 3 | Characteristic biomarkers of plant oil. Partial Total Ion Chro matograms and m ass spect ra (EI, 70eV) showing uns aturated di- and
triacylg lycerols (DAGs and TAGs).
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Article
Extende d Data Fig. 4 | TI C showing the mo lecular con stituents o f castor oil
and poss ibly other pla nt oils (blue), oil /tar of cedar oi l/tar (brown) an d
juniper/cypre ss (purple) in a w hite beaker la belled “he ad”. Cx:y = fatty a cid
with x carb on atoms and y repre senting the n umber of unsat uration;
OH-C = hydroxy fa tty acid; diOH -C = dihydroxy fatt y acid; Ric-OH = r icinoleic
acid. AID B-5 = arithme tic retentio n index.
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Extended Data Fig. 5 | Bitumen biomarkers. Ion extract chromatogram
(m/z 191 and 217) show ing a the hopane s and b the sterane s. Hopanes = T M:
17α(H), 22,29,30 -trisnorhop ane; H29: 17α(H),21β(H)- norhopane (C29);
H30: 17 α (H),21β (H)-hopane (C30); H31S: 2 2S-30-homoho pane (C31);
H31R: 22 R-30-homohopane (C31); GCR : gammacera ne; H32S: 22S-30, 31-
bishomoho pane (C32); H32R: 22R-30,31-b ishomohopane (C32); H33S:
22S-30,31,32-trishomohopane (C33); H33R: 22R-30,31,32-trishomohopane
(C33); H34S: 22 S-30,31,32 ,33-tet rakishomo hopane (C34); H34R : 22R-30, 31,3 2,
33-tetrakishomohopane (C34); H35S: 22S-30,31,32,33,34-pentakishomohopane
(C35); H35R: 22R-30,31,32,33,3 4-pentakishomohopane (C35). Steranes =
C27Sαα α: 20S-5α(H),14α(H),1 7α(H)-cholesta ne (C27); C27Rαβ β: 20R-5α(H),
14β(H),17β(H)-ch olestane (C27 ); C27Sαββ: 20 S-5α(H),14β(H),17β(H)-
cholest ane (C27); C27Rα αα: 20R-5α(H),14α(H),17α(H)-c holestane (C27);
C28S ααα: 20S-5 α(H),14α(H),17α(H)-ergosta ne (C28); C28Rαβ β: 20R-5α(H),
14β(H),17β(H)-er gostane (C28); C2 8Sαββ: 20 S-5α(H),14β(H),17β(H)-er gostane
(C28); C28Rα αα: 20R-5α(H),14α(H),17α(H)- ergostan e (C28); C29Sαα α: 20S-5α(H),
14α(H),17α(H)-stig mastane (C29); C29Rα ββ: 20R-5α(H),14β(H),17β(H)
-stigmastane (C29); C29Sαββ: 20S-5α(H),14β(H),17β(H)-stigmastane (C29);
C29Rααα : 20R-5α(H),14α(H),17α(H)-stig mastane (C2 9).
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Article
Extende d Data Fig. 6 | Saq qara complex a nd location o f the investigat ed vessels i n the embalmi ng facilitie s and burial c hambers . Background = di gital
document ation of Saqqa ra complex. Copyr ight M. Lang , Universität B onn.
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Extende d Data Fig. 7 | Pho tograph of Sha ft M23-I I (left side). Em balming worksh op/cachette roo m with ledge-like be d and drainage cha nnel, looking e ast.
Copyright SSTP.
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Article
Extende d Data Fig. 8 | Pho tograph of Sha ft M23-I I (right side). Emba lming workshop/cac hette room wi th the ledge-lin e bed and the large fu migation vess el on
the right si de, looking ea st. Copyrig ht SSTP.
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Extended Data Table 1 | List of samples including the vessels’ inscription, ORA results and quantiication
Vessels are currently stored in the Deposit room-magazine at the site of Saqqara, Egypt. TLE = Total lipid extract/g of ceramic.
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Article
Extended Data Table 2 | Synthesis of the triterpenoid components of Burseraceae resins and comparison with the products
identiied in Saqqara
Numbers correspond to references listed in theSupplementary information. BioM = biomarkers and OxM = oxidation markers.
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Extended Data Table 3 | Modern and aged angiosperm resins used as references
List of modern and aged (30 years old) angiosperm resins which have been analysed under similar conditions for comparison with the archaeological samples.
Content courtesy of Springer Nature, terms of use apply. Rights reserved

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