Food Research International 38 (2005) 1125–1134

www.elsevier.com/locate/foodres

Characterization of the edible bird’s nest the “Caviar of the East”

¤
Massimo F. Marcone
Department of Food Science, Ontario Agricultural College, University of Guelph, Guelph, Ont., Canada N1G 2W1

Received 24 January 2005; accepted 18 February 2005

Abstract

A few species of swiXets (genus Aerodramus) build edible nests that are consumed by humans worldwide, as a delicacy known as
the “Caviar of the East” or as a medicinal food. This study reports on the compositional properties of two types of nest, the white
nest and the red “blood” nest. The order of composition (from lowest to highest) was found to be identical for both types of nests,
i.e., lipid (0.14–1.28%), ash (2.1%), carbohydrate (25.62–27.26%) and protein (62–63%). It was also found that both nests share a com-
mon 77 KDa protein that has properties similar to those of the ovotransferrin protein in eggs. This protein may be partially respon-
sible for the severe allergic reactions that sometimes occur among young children who consume edible bird’s nest products. It was
found that SDS–PAGE electrophoretic Wngerprinting might serve as a useful analytical technique for diVerentiating between white
and red nests and for determining if the more expensive “blood” nest was adulterated with the less expensive white nest. Also evalu-
ated were diVerent analytical methodologies for detecting adulterants. Three of the most common adulterants found in retail bird’s
nests are karaya gum, red seaweed, and tremella fungus, and they are routinely incorporated during commercial processing prior to
Wnal sale. Using crude protein determination, it was found that these adulterants (which typically accounted for 2–10% of the
Wnished nest), reduce the overall crude protein content of the genuine white bird’s nest by as much as 1.1–6.2%. A modiWed xantho-
proteic nitric acid test for proteins proved to be a rapid, and simple test to detect adulteration in both whole and Wnely ground nests,
and would be suitable in the Weld where analytical facilities are not readily available.
After simple nitric acid treatment, visual examination and comparison of whole nests adulterated with karaya gum, red seaweed,
and Tremella fungus against the authentic white nest revealed that levels of adulteration as low as 1.7%, 1.8%, and 3.5%, respectively,
could be identiWed visually. In the case of Wnely ground nests, the visual detection level was higher for all three adulterants: 1.1% for
karaya gum, 1.2% for red seaweed, and 2.0% for Tremella fungus. The use of a reXectance colourmeter rendered this test even more
sensitive, allowing detection at even lower levels.
 2005 Elsevier Ltd. All rights reserved.

1. Introduction
“Edible bird’s nest” refers to the nest produced by
several diVerent swiftlet species. Human consumption of
these nests has been a symbol of wealth, power, and
prestige, as well as being used medicinally in traditional
Chinese medicine dating as far back as the Tang (618–
907 AD) and Sung (960–1279 AD) dynasties (Koon &
Cranbrook, 2002).
More than 24 species of insectivorous, ecolocating
swiXets are distributed around the world, but only a few
¤
Tel.: +1 519 824 4120x58334; fax: +1 519 824 6631.
produce nests that are deemed ‘edible’ (Koon, 2000). The
majority of edible bird’s nests traded worldwide come
from two heavily exploited species, the White-nest
swiXet (Aerodramus fuciphogus) and the Black-nest
swiXet (Aerodramus maximus), whose habitats range
from the Nicobar Islands in the Indian Ocean to sea-
caves in the costal regions of Thailand, Vietnam,
Indonesia, Borneo and the Palawan Islands in the Phil-
ippines (Koon, 2000; Koon & Cranbrook, 2002).
The nests are built almost exclusively by the 7–20 g
male swiXet over a period of approximately 35 days. The
building material is composed almost entirely of a gluti-
nous material found in saliva secreted from the swiftlet’s

0963-9969/$ - see front matter  2005 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodres.2005.02.008
1126 M.F. Marcone / Food Research International 38 (2005) 1125–1134
two sublingual salivary glands (Goh et al., 2001). The
half-bowl, self-supporting shaped nests, weigh 1–2 times
the swiXet’s actual body weight and are usually attached
to the vertical concave face walls of inland or seaside
caves (Fig. 1A and B). Harvesting of the edible bird’s
nest for human consumption is a painstaking and often
times dangerous operation for local collectors. Most
nests are built hundreds of feet up on cave walls and
require the use of temporary scaVolding made of locally
collected bamboo or ironwood. After collection, the
tedious process of cleaning approximately 10 nests takes
a person approximately 8 h (Koon & Cranbrook, 2002).
The nests are cleaned by soaking them in water until the
nest cement is softened and the tightly bound laminae
partially loosen. Small feathers and Wne plumage are
then manually removed with tweezers with the cleaned
strands subsequently being re-arranged and molded into
chips of various shapes, air-dried, and packaged for sale
around the world (Koon & Cranbrook, 2002) (Fig. 1C
and D). Edible bird’s nest is not the only commercially
available food product highly esteemed/priced for
human consumption processed through an animal but
also includes argan oil made from the argan nut that has
passed through the digestive track of a goat and Kopi
Luwak [the most expensive and rarest beverage/coVee in
Fig. 1. (A) Edible red “blood” bird’s nest (unprocessed); (B) edible
white bird’s nest (unprocessed); (C) edible red “blood” bird’s nest
(processed); (D) edible white bird’s nest (processed).
the world which passes through the GI track of a civet
(Marcone, 2004)].
Hong Kong is considered the world’s largest importer
and consumer of the processed nests with North Amer-
ica being the second largest market (Goh et al., 2001).
World trade Wgures conservatively estimates that 17–20
million nests are harvested each year with the total
weight being estimated at approximately two (2) metric
tonnes (Goh, Chew, Shek, & Lee, 1999; Goh et al., 2001).
After personal observation of trade practices in both
Malaysia (Borneo) and Indonesia, the above stated val-
ues appear to be under estimates of actual world wide
production.
Often referred to as the “Caviar of the East”, the nest
retails for anywhere from $2000.00 (for white nests) to
$10,000.00 (for “red blood” nests) Canadian per Kilo-
gram (Koon & Cranbrook, 2002) depending upon their
grade. They are usually prepared for consumption by
cooking them in a double boiler with sugar producing a
gastronomic delicacy often known as ‘bird’s nest soup’
which is highly esteemed as a food tonic believed to have
medicinal properties (Koon, 2000; Koon & Cranbrook,
2002). The only current scientiWc knowledge about the
medicinal properties of these nest extracts is that they
have haemogglutination inhibiting action against the
inXuenza virus (Howe, 1961; Howe, Lee, & Rose, 1960).
A more recent discovery demonstrated that partially
puriWed swiftlet nest extracts possess the Wrst known
avian epidermal growth factor (EGF) (Kong et al., 1987;
Ng, Chan, & Kong, 1986).
Unfortunately, much is still unknown about the com-
positional properties of the edible bird’s nest. It is quite
clear, however, that the salivary nest cement is the main
ingredient of the edible bird’s nest and is undoubtedly
one of the most expensive food ingredients in the world
(Koon & Cranbrook, 2002). As a result, the number of
documented and suspected cases of ‘white’ bird’s nest
adulteration with less expensive materials has risen
sharply over the past several years (Goh et al., 2001; Law
& Melville, 1994). In an eVort to increase the net weight
of the nest prior to sale, a few re-occurring materials
have consistently been identiWed as common adulterants
including karaya gum, red seaweed, and Tremella fun-
gus. These materials are usually incorporated during the
processing stages at levels approximating 10% and are
extremely diYcult to detect due to their similar color,
appearance, taste and texture to the actual salivary nest
cement.
One of the adulterants often times incorporated during
processing is karaya gum which is a dried exudation of
the stem and branches of Sterculia urens (a member of the
cacao family) and is insoluble in water. Instead, it absorbs
water forming viscous colloidal sols with adhesive type
properties similar to the nest cement. The white jelly fun-
gus (Tremella fuciformisis) is another often used adulter-
ant which when introduced in the form of thin slivers
 M.F. Marcone / Food Research International 38 (2005) 1125–1134
looks very much like the laminae strands of the edible
bird’s nest. Perhaps the most used of adulterants are the
carrageenan-bearing red seaweeds like Kappaphycus
alvarezii which is cut into slivers and boiled therefore ren-
dering them very diYcult to detect in the Wnal product.
Since little is known nor is published about the com-
position of these nests, an investigation was conducted to
elucidate if there is any substantial chemical diVerence
among ‘lower’ grade white nests and premium red
‘blood’ nests. In addition, due to the recent raise in the
rate of adulteration during processing, relatively rapid,
simple, and eVective analytical methods were explored
that could be used in the Weld for detecting possible
anomalies at much earlier stages in the distribution chain.
2. Methods
2.1. Materials
Unprocessed and processed edible white and red
“blood” birds’ nests were collected from various loca-
tions from both Malaysia (Kuala Lumpur and the island
of Borneo) and from locations on the island of Sumatra
in Indonesia. These nests comprised those collected from
both inland and seaside caves as well as man made bird
houses in both countries.
2.2. Colorimetric determination
The color (L*, a*, and b* values) of the raw edible
bird nests was determined using a Minolta Chroma
Meter CR-200b (Minolta Camera Co., Ltd., Osaka,
Japan). All analyses were performed on three indivdu-
ally collected nests of each nest type (white or ‘blood’
nests) and duplicate tests performed on each nest. Ten
individual readings were taken by placing nests perpen-
dicular to the optical sensor. All readings were averaged.
2.3. Proximate analysis of bird’s nest and elemental
(P, K, MG, and Ca) analysis
Proximate analysis was performed as prescribed in
the oYcial standard methods of the American Associa-
tion of Cereal Chemists, Inc. (AACC, 1983) after the
nests were Wnely ground using a coVee grinder (Black &
Decker, Toronto, Ont.) and screened through a 125 mm
standard sieve. For elemental analysis, 0.250 g samples
of oven-dried nest material was wet digested and sub-
jected to atomic absorption analysis as described by
Marcone and Yada (1997).
2.4. X-ray microanalysis
Uncoated samples of edible bird’s nest were attached
to carbon stubs and were scanned at a high-voltage setting
1127
of 15 kV using a Hitachi S-4500 Field-Emmission Scan-
ning Electron Microscope equipped with an “Quartz
One” energy dispersive X-ray spectrometer. X-ray system
was calibrated to measure elements from Boron and
higher. Analyses were performed essentially as described
by Houston, Moore, Favrin, and HoV (2004). All data
analysis was performed by software supplied by Hitachi.
2.5. SDS–PAGE
SDS–PAGE electrophoresis was performed on raw
nest material after normalization for diVerences in pro-
tein content. Approximately 50 mg of Wnely ground nest
material was dissolved in 1 ml of SDS–PAGE standard
derivitization and procedure run according to the
method outlined by Marcone and Yada (1997). Stan-
dard molecular weight proteins included,
-lactalbumin,
14,400 Da ovalbumin, 43,000 Da, bovine serum albumin
67,000 Da; phosphorylase b, 94,000 Da (Pharmacia,
LKB, Montreal, Canada).
2.6. Phenol–sulfuric acid reaction for carbohydrates
Total carbohydrate content was determined as
described by Dubois, Gilles, Hamilton, Rebers, and Smith,
1956. Glucose was used in preparing the standard curve
ranging from 2 to 10 g per sample. Samples were allowed
to stand until no further color change could be detected.
2.7. Amino acid analysis
Amino acid analysis was performed on the white and
red “blood” birds’ nests exactly as described by Marcone
and Yada (1997).
2.8. Fatty acid analysis
Fatty acid analysis was performed as described by
Christie (1982) with minor changes, was used to prepare
the fatty acid methyl-esters. The fatty acid derivatives
were analyzed using a Shimadzu Gas Chromatograph
(Model GC-14 A, Man-Tech. Associates Inc., Guelph,
Ont.), equipped with a split mode injection system, Xame
ionization detector (FID), and fused silica capillary col-
umn (SP-2330) with 30 £ 0.25 mm ID and 0.25 M Wlm
thickness (Supelco, Oakville, Ont.). The initial oven tem-
perature was 145 °C and temperature programmed at a
rate of 6 °C/min to 235 °C. The intial time was 2.0 min,
and the Wnal hold time was 10 min. The injection port
temperature was 260 °C, and the detector port tempera-
ture was 260 °C. The hydrogen gas and air Xow rate were
adjusted at 30 and 300 ml/min, respectively, and the car-
rier hydrogen gas rate was 1.0 ml/min. The sample
volume was 0.1 l and the data were integrated with a
Shimadzu Model C-R4A Chromatopac (Man-Tech
Associates Inc., Guelph, Ont.). The fatty acid
1128 M.F. Marcone / Food Research International 38 (2005) 1125–1134
composition was determined using standard fatty acid
methyl esters (Sigma–Aldrich, Oakville, Ont.).
2.9. Triacylglyceride analysis
The AOCS oYcial method (Ce 5b-89, 1995) after
slight modiWcation to the mobile phase ratio was used to
determine the TAG composition of extracted lipid from
bird’s nests. The separation was performed on two
Econosil C18 columns (5 m, 4.6 £ 250 mm, Alltech,
DeerWeld, IL) placed in series. The analysis was carried
out isocratically with a mobile phase consisting of
60:40% (v/v) acetone:acetonitrile (v/v). Sampes (5%)
were dissolved in HPLC grade acetone and 20 l ali-
quots were automatically injected onto the column
(Waters 700 Satellite WISP, Millipore, Milford, MA)
and eluted at a Xow rate of 1 ml/min. The column was
equilibrated at 30 °C with a sensitivity scale at 64. The
TAG were identiWed by comparing retention times to
pure standards purchased from Sigma–Aldrich
(Oakville, Ont.).
2.10. Scanning electron microscopy
Nest fragments were Wxed at 23 °C with 5% gluteral-
dehyde in 0.05 M phosphate buVer pH 7.0 for 2 h. After
10 washes in phosphate buVer, they were post-Wxed in
1.0% osmium tetraoxide in the same buVer for 1 h at
23 °C. The Wxed fragments were rinsed with phosphate
buVer and dehydration was achieved using an ethanol
series. Samples were then critical point dried using car-
bondioxide. Nest fragments were then mounted on alu-
minum stubs and coated with gold/palladium (60/40) to
a thickness of 25–30 nm using an Anatech Hummer VII
sputter coater (Alexandria, VA). Following coating, the
nest fragments were viewed under a Hitachi S-570 Scan-
ning Electron Microscope with a high voltage setting of
10–20 kV.
2.11. Xanthoproteic acid test for proteins
Whole and ground white edible birds’ nests were
tested for protein by application of three drops (approx-
imately 100 l) of concentrated nitric acid to their
respective nest material and comparison of color devel-
opment performed exactly 20 s after application. Con-
centrated nitric acid reacts with tyrosine, phenylalanine,
and tryptophan yielding red-yellow color products.
2.12. Determination of visual sensitivity in adulteration
detection
In order to determine the level of visual sensitivity for
the detection of adulterated nests using the modiWed
xanthoproteic acid test for proteins, panelists were asked
to evaluate adulterated nests in the range of 0.5–10%.
A DUO-Trio test diVerence analyses was conducted
by presenting 24 panelists (12 male and 12 females) an
unadulterated control and a pair of coded samples, one
of which was identical to the control and one adulter-
ated to a known level. The panelists were asked which
sample was diVerent. Progressingly, lower levels of each
of the three adulterants were tested to determine at
which level 95% conWdence interval of the panelists
could still pick out the adulterated sample. All samples
were treated with nitric acid just prior to presentation to
the panelist. Tests were performed in triplicate.
2.13. Statistical analysis
All analyses were performed on three individual sam-
ples unless otherwise indicated. Statistical analysis was
performed using a SAS Statistical Analysis System pack-
age. SigniWcant diVerences among samples were deter-
mined by Duncan’s multiple range test (p 6 0.05) (SAS,
1990).
2.14. Results and discussion
A structural examination of both raw and processed
white and red “blood” birds’ nests indicated that they
were composed of repeatedly interwoven strands of sali-
vary laminae cement (Fig. 1A–D). Fig. 1A and B shows
the appearance of unprocessed nests where as Fig. 1C
and D shows nests that have been processed/cleaned and
ready for commercial sale to the consumer. Overall, the
laminae strands composing both white nests and red
“blood” nests were highly variable ranging from 2 to 4
mm in diameter. The strands from each nest can be eas-
ily distinguished from one another by a visual observa-
tion of their overall color (Fig. 1). The lower grade
“white” nests had an oV-white hue while the red ”blood”
nests exhibited an overall reddish “terracotta” color. The
red color associated with the latter is neither water-solu-
ble nor lipid/solvent extractable. Unfortunately, it has
been known that on occasion white nests have been
treated with red pigments which are either partially or
wholly water-soluble so as to give the false appearance
of the nest being of higher grade (i.e., red “blood” nest),
and hence command a higher price from unexpectant
consumers.
Various tests were conducted on both types of nests
to determine their respective composition. The order of
composition (from lowest to highest) was found to be
identical for both types of nests, i.e., lipid, ash, carbohy-
drate, and protein (Table 1).
Test results from both the white and red nests indi-
cated that lipids constituted the smallest measured frac-
tion but also that there were diVerences between the
white and red nests. The white nest contained only min-
ute trace amounts of lipid, i.e., approximately 0.14%
whereas the red nest contained approximately 1.28% or 9
 M.F. Marcone / Food Research International 38 (2005) 1125–1134 1129

Table 1
Composition analysis of edible birds’ nests and common adulterants
White bird’s Red “blood” Karaya gum Red seaweed Tremella fungus
nest bird’s nest
Proximate analysis (%)
Moisture 7.50a 8.00b 17.52c 44.63d 4.50a
Ash 2.10a 2.10a 16.11b 33.94d 7.64c
Fat 0.14a 1.28b 1.20b 2.32c 2.22c
Protein 62.0d 63.0d 0.70b 0.40a 8.60c
Carbohydrate 27.26c 25.62b 74.47d 18.71a 77.04e
Elemental analysis (ppm)
Sodium (Na) 650c 700a 70a 50,350d 180b
Potassium (K) 110a 165b 6900b 31,640e 26,440d
Calcium (Ca) 1298c 798b 1642d 1840d 190a
Magnesium (Mg) 330b 500a 3040c 6100d 520a
Phosphorus (P) 40b 45b 10a 90c 4060d
Iron (Fe) 30c 60d 10a 20b 20b
Fatty acid analysis (%)
(P) Palmitric C16:0 23a 26a
(O) Steric C18:0 29a 26a
(L) Linoleic C18:1 22a 22a
(Ln) Linolenic C18:2 26a 26a
Triacylglycerol (%)
PPO 16a 14a
OOL 13a 15a
PLnLn 19a 18c
Monoglycerides 31b 27a
Diglycerides 21a 26a
Values in category row with the same letter are not signiWcantly diVerent (p 7 0.05).

times more than the white nest (Table 1). This prelimi-
nary result could possibly be another deWnitive test to
diVerentiate between the two nest types. Further exami-
nation of this lipid component revealed that each nest
type had similar mole fractions (i.e., percentages) espe-
cially those fatty acids of nutritional signiWcance, namely
palmitic C16:0 (P), stearic C18:0 (O), linoleic C18:1 (L),
and C18:2 (Ln). Interestingly, triacylglycerol analysis
revealed the presence of over 52% mono- and diglcerides
with equimolar ratios of PPO, OOL, PLn Ln composing
the rest of the TAG present. The origin or function of
such high content of mono- and diglycerides is not yet
understood. It may be that the extremely high relative
humidity of the caves (>80%) together with surface con-
densation may have lead to hydrolytic cleavage of the
triacylglyerols. The action of enzymes found in the bird’s
saliva needs also to be taken into consideration. Further
investigation into the area will be required especially on
freshly made nests before any deteriation can occur to
them.
The amount of inorganic ash in both nests was virtu-
ally identical at 2.1% (Table 1). Although the total
amount of minerals from each nest were identical,
atomic absorption analysis indicated signiWcant diVer-
ences in the total amounts of various nutritionally
important minerals, namely calcium, potassium, magne-
sium and iron. White nests were found to be much
higher in calcium as were the red nests which were found
to be much higher in potassium, magnesium and iron.
All red nests tested were found to have typically higher
levels of iron.
The results of the carbohydrate analysis of both white
and red nests are presented in Table 1. The test results
indicate that this was the second highest occurring compo-
nent in all of the nests with some diVerences, notably that
the lower-grade white nests had slightly more total carbo-
hydrate content than the higher-grade red “blood” nests.
By far the most abundant component in both nest
types was crude protein with the red “blood” nests hav-
ing a slightly higher but not signiWcantly higher
(p 7 0.05) level (63%) compared to the white nests (62%).
Amino acid analysis also revealed that both nest types
had very similar amino acid proWles that were rich in cer-
tain, but not all essential amino acids (Table 2).
To learn more about the types of proteins that consti-
tute this particular dominant fraction within the respec-
tive nests, SDS–PAGE electrophoresis was performed.
The results demonstrate that each nest type possesses its
own unique protein Wngerprint, indicating that each is
composed of diVerent protein species (Fig. 2), suggesting
that not only could this analytical procedure potentially
serve as a method to diVerentiate between these two
diVerent nests, but also as a method of determining the
identity and/or purity of a nest for possible regulatory
purposes. The staining intensity of their constituent
protein bands might also be useful in assessing if an
1130 M.F. Marcone / Food Research International 38 (2005) 1125–1134

Table 2
Amino acid determination of white and red “blood” edible birds’ nests
Amino acidA White edible Red “blood”
bird’s nest edible bird’s nest
Aspartic + asparagines 9.5 10.0a
Threonine 4.4b 5.3b
Serine 15.4a 15.9a
Glutamic + glutamine 7.0a 7.0a
Glycine 5.9a 4.8b
Alanine 4.0a 4.7b
Valine 10.7a 11.1a
MethionineB 0.8a 0.0b
Isoleucine 10.1a 10.7a
Leucine 3.0a 3.4b
Tyrosine 10.1a 9.0b
Phenylalanine 6.8a 6.7a
Lysine 3.5a 2.8b
Histidine 3.3a 2.6b
Arginine 5.4a 6.1b
Values in category row with the same letter are not signiWcantly diVer-
ent (p 7 0.05).
A
Molar percent basis.
B
By oxidation.

adulteration of the premium red “blood” nest with the

lower grade “white” nest has occurred. In addition, areas
around the protein bands were observed to be sur-
rounded by substantial streaking, which is commonly
observed for highly polymerized protein complexes
(Fig. 2). Furthermore, both nest types share one band
with a molecular weight of approximately 77 kDa, the
protein being glycosylated to approximately the same
extent for each type (i.e., 2.7%).
The molecular weight of the common protein in both
nest types had a strong band at 77 KDa that is very sim-
ilar in molecular weight to the highly allergenic ovo-
transferrin protein in eggs. It has been demonstrated that
ovotransferrin is glycosylated (Williams, Elleman,
Kingston, Wilkins, & Kuhn, 1982) and, interestingly, it is
glycosylated to the same level as the 77 kDa protein
band in the nests (2.8%). It is still unclear, however, if an
ovotransferrin-like protein is synthesized elsewhere
other than in eggs but these tentative results demon-
strate that there is some semblance of similarity between
the protein in eggs and that found in the birds’ nests.
Further amino acid sequence studies will help to further
elucidate how these proteins are inter-related. Interest-
ingly, the red (terracotta) color of the “blood” nest is
very similar to the color of puriWed ovotransferrin in its
iron complexed state whereas the “white” colored nest is
similar in color to ovotransferrin in its non-iron com-
plexed form. This also supports the proposed theory that
the protein in both nest types is at least ovotransferrin-
like, very similar to ovotransferrin in eggs. Further sup-
porting this theory is the diVerence in the iron contents
of both nest types as indicated by atomic absorption
analysis. Scanning electron microscopy combined with
X-ray microanalysis performed at various locations on
Fig. 2. (I) SDS–PAGE (reducing 1 l of 1 mg¡1 protein solutions)
applied to Homogenous 20 Phast gels (Pharmacia LKB). (lanes S)
standard [
-lactalbumin, 14,400 Da; soybean trypsin inhibitor, 20,100
Da; carbonic anhydrase, 30,000 Da; ovalbumin, 43,000 Da; bovine
serum albumin, 67,000 Da; phosphorylase b, 94,0000 Da] (1), ovo-
transferrin; (2), total protein – red ‘blood’ bird’s nest; (3), total protein
– white bird’s nest. (II) Densitometric scan of gel (all labels are the
same as in (I).
the surface of the red nests indicate relatively higher lev-
els of iron compared to that of the white nests. These
analyses also indicated that the distribution of various
elements in the nests are highly variable indicating possi-
ble compositional diVerences in the secreted saliva dur-
ing the 35-day period when the nests are being built
(Fig. 3).
Since proteins constitute the major compositional
fraction of both studied nest types, it is not uncommon
that several medical studies have shown the occurrence
of a severe allergic reaction among some young children
who have consumed ‘bird’s nest soup’ (Goh et al., 2001).
The symptoms of these allergic reactions were similar to
those induced by egg-like proteins. Interestingly, these
studies also noted that bird’s nests-induced allergies
were the most common cause of anaphylaxis reported at
the National Hospital in Singapore even surpassing
other well-deWned food allergens such as cow’s milk or
egg in younger children and peanut or crustacean sea-
food in older children (Goh, Chew, Chua, Chay, & Lee,
2000; Goh et al., 1999). In North America as well as the
 M.F. Marcone / Food Research International 38 (2005) 1125–1134 1131

Fig. 3. Scanning electron microscopy combined with X-ray microanal-

ysis of red “blood” edible bird’s nest. (A) X-ray microanalysis of “red”
nest portion. (B) X-ray microanalysis of “white” nest portion.

rest of the world, however, there is no or very little infor-

mation as to how many children have experienced bird’s
nest-induced anaphylaxis. The medical community may
not even be aware of its existence or properly trained to
diagnose it.
Practitioners of traditional Chinese medicine have
consistently indicated that the consumption of birds’
nest soup is beneWcial for the treatment of a variety of
health problems (Koon & Cranbrook, 2002). It is often
administered to the elderly and very young alike who are
recovering from various types of infections. Studies have
indicated that ovotransferrin is both bacteriostatic and
bacteriocidal in nature (Ibrahim, Sugimoto, & Aoki,
2000). If the protein identiWed in these nests is in fact an
ovotransferrin-like protein, then it could be reasonably
involved in or responsible for Wghting infection just like
the ovotransferrin in eggs. This may be a possible reason
as to why edible bird’s nest soup has been consistently
consumed throughout the centuries as prescribed tradi-
tional Chinese medicine.
The salivary nest cement is the most important ingre-
dient in the edible bird’s nest and is undoubtedly one of Fig. 4. (A) Red seaweed; (B) karaya gum (plant resin); (C) Tremella
the most expensive food ingredients in the world. Often fungus.
times, however, it has been found adulterated with
Tremella fungus, karaya gum or red seaweed (Figs. 4 and weed were easily distinguishable from strands of birds’
5). These three materials are typically only incorporated nest cement. Cross-sections of seaweed strands con-
to a maximum of 10% to increase the overall net weight tained very detailed striations on raised surfaces which
of the Wnished nest. These compounds are ideal adulter- were distinguishable from the high amophorous strands
ants since their color, taste, and texture are similar to of bird’s cement (see inset of micrograph; Fig. 6).
that of the genuine birds nests salivary cement and are Compositional analysis of these common adulterants
therefore often diYcult to detect. revealed that they contain minerals in diVering amounts
Scanning electron microscopy did not prove to be an and ratios rendering each with a unique and distinguish-
eVective technique for determining the existence nor the able mineral proWle (Table 1). Since iodine and salt
type of adulteration in edible bird’s nest except for those (NaCl) are present in red seaweed, but are not typically
containing added red seaweed. Unlike these adulterated found in genuine birds nest, presumptive tests for iodine
with tremella fungus or plant resin, particles of red sea (iodine spot test) and salt (silver nitrate spot test) could
1132 M.F. Marcone / Food Research International 38 (2005) 1125–1134
Fig. 5. (A, C, and E) Edible white bird’s nest (processed); (B) edible red
seaweed adulterated white bird’s nest (processed); (D) edible karaya
gum (plant resin) adulterated white bird’s nest (processed); (F) edible
tremella fungus adulterated white bird’s nest (processed).
quickly and easily be used to detect non-heat treated sea-
weed adulterants (results not shown). Unfortunately,
most violators heat treat their seaweed prior to adultera-
tion, decreasing both the iodine and NaCl (to the same
extent) levels and thereby limiting detection test usage.
Nests containing abnormally high potassium compared
to pure birds nest material could indicate adulteration
with red seaweed and/or Tremella fungus, which are
both relatively high in this mineral. On the other hand,
nests with abnormally high levels of phosphorous could
indicate adulteration with Tremella fungus, whereas high
amounts of sodium could indicate adulteration with red
seaweed.
Fig. 6. Scanning electron micrograph of seaweed cross-section found
in adulterate nest. Not striations in box.
Closer compositional examination of the three adul-
terants – namely karaya gum, red seaweed and Tremella
fungus – revealed that they were relatively low in overall
crude protein, i.e., 0.7%, 0.4% and 8.6%, respectively,
compared to the 62% of pure white birds nest material
(Table 1). Intentional additions of as little as 2% to as
high as 10% (with 10% being the most commonly found
level of adulteration found) substantially decreased the
overall total protein content of the nest ranging from
1.1% to 6.2%, as measured by total protein analysis.
Therefore, comparison of the total protein contents of
suspected adulterated birds nest versus the pure edible
bird’s nest could potentially be used to determine nest
authenticity.
Amino acid analysis revealed that white nest protein
was substantially high in two speciWc aromatic amino
acids, namely phenylalanine and tyrosine (Table 2). Con-
centrated nitric acid reacts with aromatic amino acids in
proteins to produce a yellow-red color in proportion to
the amount of protein present. This xanthoproteic acid
test for proteins could therefore potentially be used to
detect adulterated nests for sale on the market. All whole
adulterated nests produced a visibly less intense yellow-
red reaction when treated with nitric acid when com-
pared to the same test performed on the genuine white
nests.
When pieces (1.0 £ 1.5 cm) of nests that were artiW-
cally adulternated with karaya gum, red seaweed and
 M.F. Marcone / Food Research International 38 (2005) 1125–1134 1133

Fig. 7. (A) Whole edible red seaweed adulterated white bird’s nest xanthoproteic nitric acid test); (B) whole pure edible white bird’s nest (xanthopro-
teic nitric acid test).

Tremella fungus were presented to panelists, their detec- Table 3

tion down to 1.7%, 1.8% and 3.5%, respectively, could be Minolta colorimetric reading of ground bird’s nest with variable
amounts of adulterants treated with nitric acid (xanthoproteic acid
made. Fig. 7 shows two such nest pieces (one pure and test)
one adulterated with 5% red seaweed) treated directly
Typical adulteration level
with nitric acid. The unadulterated (pure nest) was much
darker in color than the adulterated one due to the 2% 5% 10%
higher content of protein. In the Wnely ground nests, the L* a* b* L* a* b* L* a* b*
use of this test permitted the vision detection of adultera- Seaweed 56a 48c 32a 60b 40b 37 65c 37a 42c
tion to levels of 1.1% (karaya) gum, 1.2% (red seaweed), Karaya gum 55a 50c 30a 62b 40b 35 65c 35a 40c
2.0% (Tremella fungus). Fig. 8 shows the color diVerence Tremella fungus 50a 50c 25a 55b 45b 27‘ 60c 40a 30c
Pure Nest 43 58 25
that could be expected with adulteration of 10% typi-
cally found in the market place. This may be due to the Values in each row of the same descriptor, i.e., all L*, a* or b* (sepa-
rately) with the same letter are not signiWcantly diVerent (p 7 0.05).
more uniform distribution of protein and therefore more
uniform color development in the test material. The use
of a reXectance colourmeter made this test even more cal L*, a*, b* values for adulteration reading between
sensitive down to 0.2% for karaya gum, 0.3% for red sea- 2% and 10% which are typical ranges found in the mar-
weed, and 0.5% for Tremella fungus. Table 3 shows typi- ket place.

Fig. 8. (A) Crushed edible karaya gum adulterated white bird’s nest (10% adulteration); (B) crushed edible white bird’s nest (10% adulteration) (all
treated with nitric acid – xanthoproteic nitric acid spot test).
1134 M.F. Marcone / Food Research International 38 (2005) 1125–1134
In conclusion, this study reports on the actual compo-
sitional properties of both white and red “blood” nests
revealing that they both contain predominately protein-
ous material followed by carbohydrate, ash and lipid (in
descending order). SDS–PAGE was found to be an ade-
quate analytical method to diVerentiate between these
two nest types whereas a modiWed xanthoproteic acid
test was found to facilitate the diVerentiation detection
of “whote” nests that were adulterated with commonly
used materials.
When pieces (1.0 £ 1.5 cm) of nests artiWcally adulter-
ated with karaya gum, red seaweed and Tremella fungus
were tested, panelist could detect their presence down to
1.7%, 1.8% and 3.5% inclusion, respectively. Fig. 7 shows
two nest pieces directly treated with nitric acid and
shows the diVerence in color between adulterated and
pure nests.
Acknowledgements
The author thank Dr. Anthony Clarke of the Depart-
ment of Microbiology for performing amino acid analy-
sis and Dr. Alexandra Smith of the Department of Food
Science for her assistance in X-ray microanalysis.
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