Mal J Nutr 5:55-59, 1999
The potential of the coelomic fluid in sea cucumber as an antioxidant
Hawa I, Zulaikah M, Jamaludin M, Zainal Abidin AA, Kaswandi MA and Ridzwan BH
Department of Biomedical Science, Faculty of Allied Health Sciences,
Universiti Kebangsaan Malaysia, Jalan Raja Muda A. Aziz, 50300 K. Lumpur, Malaysia.
In a biological system the product of free-radical induced-lipid peroxidation are
conjugated dienes (CDs), lipid peroxides and malondialdehyde (MDA). Its control
depends on antioxidant activities. Antioxidant defend the body system by controlling
the damage caused by the free radicals. This paper describes lipid peroxidation process
and antioxidant activities present in three species of holothuroid obtained from the
coastal waters of Terengganu, Malaysia. Among the three species studied, the
coelomic fluid of Bohadschia mamorata vitiensis contained the highest level of
protein (7.24 + 0.04)g/L. The coelomic fluid of Stichopus badionotus Selenka
contained the highest level of superoxide dismutase (SOD) (9.46 + 0.99)X 105 IU/g
protein, MDA (16.46 + 1.28)nmol/g protein and total antioxidant activities (AOA)
(58.81 + 5.70) %. In Stichopus variegatus Semper the coelomic fluid only
demonstrated the highest level of CDs (94.19 + 11.34) RU/g protein compared with
the other two species. This preliminary study indicated that some form of antioxidant
activities are present in the coelomic fluid of holothuroids.
In the south-east Asia regions sea
cucumbers are taken as a food supplement
(Ridzwan and Che Bashaah 1985; Ridzwan
1993). Holothuroid and its product could cure
certain ailments (Shimada 1969; Sit 1998).
Fishermen often consume the liquid portion
or the coelomic fluid of sea cucumbers in an
attempt to remain healthy for days while out
at sea. It is believed that if consumed
regularly sea cucumbers could reduce
hypertension, asthma, heal internal wounds
and cancer. This bentoic organism has a
vascular system filled with fluid believed to
contain bioactive substances important for
wound healing (Perchenik 1969).
56 Hawa I, Jamaludin M, Zainal A, et. al.
Certain extracts of sea cucumbers had
also been described to contain vitamin E, an
antioxidant (Madhavan 1998). Therefore, the
purpose of this study is to further investigate
whether other antioxidant activities are
present in the coelomic fluid of three species
of holothuroids that inhabited the Malaysian
coastal waters. The species investigated are
Bohadschia mamorata vitiensis, Stichopus
badionotus Selenka and Stichopus variegatus
MATERIALS AND METHODS
Samples were collected from the coastal
waters of Terengganu, Malaysia. An incision
measuring 3-5 cm were made on the ventral
side taking care not to injure the internal
organs. The coelomic fluid were collected in
separate beakers, later transferred into test
tubes, frozen until analysed. The protein
component was measured by using the Biuret
calorimetric method (Wooton and Freeman
1982), while the MDA levels determined
using the standard procedures (Hunter and
Jamaludin 1986). The activity of SOD was
determined by a modified Flohe and Otting
(1984) method, whereas total antioxidant
activity was measured according to Stock et
al. (19874). Conjugated dienes was
determined following the Lunec and
Dormandy (1978) method.
The results of the antioxidant activities
in the coelomic fluid of the three species of
holothuroids is shown in Table 1. The
coelomic fluid of Bohadschia mamorata
vitiensis contained the highest concentration
of crude protein (7.24 + 0.04 g/L) followed
by Stichopus variegatus Semper (2.05 + 0.40
g/L) and Stichopus badionotus Selenka (1.80
+ 0.24 g/L).
Among the three species studied the
highest level of SOD activity was detected in
the coelomic fluid of Stichopus badionotus
Selenka (9.40 + 0.99)X 105 IU/g protein
followed by Stichopus variegatus Semper
(8.20 + 0.91) X 105 IU/g protein and
Bohadschia mamorata vitiensis (5.30 + 0.18)
X 105 IU/g protein.
The coelomic fluid also contained CDs,
the product of primary lipid peroxidation. It
was the highest in Stichopus variegatus
(94.19 + 11.34) RU/g protein followed by
Bohadschia mamorata vitiensis (78.71 + 1.56)
RU/g protein and Stichopus badionotus
Selenka (70.00 + 4.55) RU/g protein.
Stichopus badionatotus contained the highest
level of (16.46 +1.29) nmol/g protein,
followed by Stichopus variegatus (9.89 +
0.04) nmol/g protein and Bohadschia
mamorata vitiensis (7.41 + 0.86) nmol/g
protein. Total antioxidant activity was found
to be the highest in Stichopus badionotus
Selenka (58.81 + 5.70) %, followed by
Bohadschia mamorata vitiensis (52.20 + 5.70)
% and Stichopus variegatus Semper (47.71 +
The potential of coelomic fluid in sea cucumber 57
TABLE 1. The Composition of Sea cucumber Coelomic Fluid
Species Product Amount
Stichopus badionotus Selenka Crude protein 1.80 + 0.24 g/L
Superoxide dismutase 9.40 + 0.99 X 105 IU/g protein
Malondialdehyde 16.46 + 1.29 nmol/g protein
Conjugated dienes 70.00 + 4.55 RU/g protein
Total antioxidant activity 8.81 + 5.7 %
Stichopus variegatus Semper Crude protein 2.05 + 0.40 g/L
Superoxide dismutase 8.20 + 0.91 X 105IU/g protein
Malondialdehyde 9.89 + 0.04 nmol/g protein
Conjugated dienes 94.19 + 1134 RU/g protein
Total antioxidant activity 47.71 + 8.95 %
Bohadschia mamorata vitiensis Crude protein 7.24 + 0.04 g/L
Superoxide dismutase 5.30 + 0.18 X 105IU/g protein
Malondialdehyde 7.41 + 0.86 nmol/g protein
Conjugated dienes 78.71 + 1.56 RU/g protein
Total antioxidant activity 52.20 + 5.70 %
Sea cucumbers, as an alternative source
of natural dietary antioxidants probably plays
an important role to counter the detrimental
effects of oxygen free radicals formed from
normal metabolism and external factors
(pollution, radiation). This is implicated in the
development of today’s common illness such
as cancer (gene damage), premature ageing,
cardiovascular and other degenerative
diseases. Indeed, our study revealed the
presence of a mild concentration of total
protein in the coelomic fluid of all the three
species of holothuroid. The findings,
however, are similar to the results reported by
Canicatti (1989, 1990) but on Holothuria
polii and Marthasterias glacialis.
Furthermore, our study also showed a
remarkable increase in the activity of
antioxidant enzymes, namely, SOD. This is in
agreement with our study on lipid
peroxidation of polyunsaturated fatty acids of
ox brain homogenates as a bioassay system
for auto oxidation. A similar result was
obtained when using plasma serum incubated
in tissue homogenate inhibit lipid auto
oxidation process (Barber 1961). According
to Dormandy (1969) all animal tissues have a
natural defence system to inhibit the process
of oxidation towards polyunsaturated fatty
acid either by enzymatic or non-enzymatic
reaction. The main role of antioxidants invivo
is in the prevention of the peroxidation of
PUFAs and the stabilization of the biological
However, the role of this antioxidant in
holothuroids remain to be elucidated. Most
probably, the increased production of oxygen
derived free radicals during hypoxic alteration
due to external changes
58 Hawa I, Jamaludin M, Zainal A. et. al.
occurred at a highly accelerated rate, that is,
when molecular oxygen was reintroduced to
the previously hypoxic tissues of
holothuroids. Thus, hypoxia enhanced the
damages induced by oxygen metabolites
complicated with the reintroduction of
molecular oxygen when the antioxidant
defense system of holothuroid was not
effective or overloaded. Thus, our findings
are similar to previous reports on human
plasma (Barber 1961; Vidlakova et al. 1972;
These results indicated that the
coelomic fluid of holothuroids from
Terengganu coastal areas contained
antioxidant substances. These substances
could possibly be produced during the
hypoxic re-oxygenation, during which a high
level of protection against the oxygen induced
damage of “stunned holothuroid” after being
exposed to the polluted habitat. It also appears
that all animals have some antioxidant
activities but the strength varies with species.
Therefore, holothuroids could provide a good
source of external antioxidant for human in
The study is funded by IRPA grant 09-
02-02-0021. “Identification and evaluation of
structure, bioactive agent and consumption of
local sea cucumbers”.
Barber, A.A. 1961. Inhibition of lipid
peroxide formation by vertebrate blood
serum. Arch. Biochem. Biophysics. 96:30-
Canicatti, C. 1989. Tthe lytic system of
Holothuria polii (Echinodermata) : A
review. Boll. Zool. 55:139-144.
Canicatti, C. 1990. Protease activity in
Holothuria polii coelomic fluid and
coelomocyte lysate. Comp. Biochem.
Dormandy, T.L. 1969. Biological
rancidification. Lancet. 27(2) 2722.
Flohe, L. and Otting, F. 1984. Oxygen
radicals in biological system.(Part A :
Oxygen radical and antioxidants). Method.
Enzymology 105: 93-97.
Hunter, M.I.S. and Jamaludin, M. 1986.
Plasma antioxidants and lipid peroxidation
product in Duchenne muscular dystrophy.
Clin. Chim. Act. 155:123-132.
Lunec, J. and Dormandy, T.L. 1978.
Flourescent lipid-peroxidation products in
synovial fluid. Clin. Sci. 56:53-59.
Madhavan, R. K.1998. Mengesan kehadiran
vitamin E dan unsur-unsur logam dalam
dalam tiga spesies timun laut. Tesis Sarjana
Muda Sains Bioperubatan (Kepujian). K.
Lumpur : Fakulti Sains Kesihatan
Bersekutu, Universiti Kebangsaan Malaysia.
The potential of the coelomic fluid in sea cucumber 59
Perchenik, J.A. 1996. The Echinoderms. In
Dubuque W.M.C Biology of Invertebrates.
3rd ed. 445-474. London : Brown Publisher.
Ridzwan, B.H. 1993. Ekinoderma yang boleh
dimakan: holothuroid: Dlm. Sumber
Makanan persisiran laut Sabah. 149-188.
Kuala Lumpur : Dewan Bahasa dan Pustaka.
Ridzwan, B.H. and Che Bashah,C.C. 1985.
Tinjauan awal taburan dan penilaian
holothurian di beberapa kawasan perairan
Sabah sebagai sumber sumber pemakanan.
Sumber, 1:143-155. Fakulti Sains Sumber
Alam, Universiti Kebangsaan Malaysia.
Shimada, S. 1969. Antifungal steroid
glycoside from sea cucumber. Science.
Sit, N.W. 1998. Aktiviti antifungus timun laut
terhadap yis. Tesis
Sarjana Muda Sains Bioperubatan
(Kepujian). K. Lumpur : Fakulti Sains
Kesihatan Bersekutu, Universiti Kebangsaan
Slater, T.F.1972. In: Free radical mechanism
in tissue injury. London. Pion
Stock., J. Gutteridge, J.M.C. Sharp, R. J.
Dormandy, T.L.1974. The inhibition of lipid
autoxidation by serum and its relation to
serum protein and alpha-tocopherol. Clin.
Sci. Mol. Medic. 47:223-233.
Vidlakova, M. Erazimova, J. Horka, J. and
Placer, Z. 1972. Relationship of serum
antioxidant activity to tocopherol and the
serum inhibition of lipid peroxidation. Clin.
Chem. Acta. 36:61-66.
Wootoon, I.D.P. and Freeman, H. 1982.
Proteins, microanalysis in medical
biochemistry 6th edition. New York: