Agriculture Science Developments, 2(4) April 2013, Pages: 31-39

TI Journals
ISSN
2306-7527

Agriculture Science Developments

www.tijournals.com

Determination of Chemical Compositions of Snail-Eating

Turtle (Malayemys macrocephala) Eggs
Tanasorn Tunsaringkarn *1, Wanna Tungjaroenchai 2, Wattasit Siriwong 3
1,3
2

College of Public Health Sciences, Chulalongkorn University, Bangkok 10330, Thailand.

Faculty of Agro-Industry, King Mongkuts Institute of Technology Ladkrabang, Bangkok 10520, Thailand.

AR TIC LE INF O

AB STR AC T

Keywords:

This study was designed to determine the chemical compositions and profiles of amino acid,fatty
acid, vitamin, mineral and sex hormone contents of snail-eating turtle eggs. Average proximate
analyses of weight, ash, moisture, carbohydrate, protein, fat and total calories of the snail-eating
turtle whole eggs were 11.55 g egg-1 , 1.45 g 100g-1 , 72.93 g 100g-1 , 4.64 g 100g-1, 12.42 g 100g-1,
8.56 g 100g-1, 145.17 kcal 100g-1 respectively. Most of these nutrients were higher in egg yolks
than egg white except moisture. Most of non-essential fatty acid (NEFA) was oleic acid (2.65 mg
100g-1) while most of essential fatty acid (EFA) was arachidonic acid (0.47 mg 100g-1). Snaileating turtle eggs contained high ash (1.45 g 100g-1 ) which was related to mineral content. Our
results also revealed that whole eggs had high essential mineral of nitrogen and trace mineral of
iron and zinc, which most of them were in egg white, In addition, it contained high vitamin A
(retinol) and E in egg yolk. Most of sex hormones content, such as Luteinizing Hormone (LH),
Follicle Stimulating Hormone (FSH), Prolactin (PRL), Progesterone (P), Estradiol (E2) and
Testosterone (T), were higher in egg yolk than in egg white, with the highest of progesterone. This
study indicated that snail-eating turtle eggs contained high contents of some essential amino acid,
such as arachidonic acid and linoleic acid, vitamin A, total minerals of nitrogen, iron and zinc and
sex hormone progesterone.

Snail-eating turtle egg

Chemical compositions
Whole egg
Egg White
Egg yolk

2013 Agric. sci. dev. All rights reserved for TI Journals.

1.

Introduction

Eggs have been consumed by mankind for millennia which are many different species, including birds, reptiles, amphibians, and fishes.
They have the various nutrients such as proteins, fats, carbohydrates, vitamins and minerals [1-3] which are the major energy sources and
provide a range of essential nutrients for tissue developments and functions [4-6]. Moreover, eggs are also an inexpensive single-food
source of proteins. Generally, c and quail eggs are the most commonly consumed eggs. However in Thailand, people who live near
agricultural areas consume the snail-eating turtle eggs which they believed that snail-eating turtle eggs are good nutrient supplements and
enhance the male sex hormone. Snail-eating turtle eggs can be found in all lowland areas of Chao Phraya River Basin [7]. Thus, this study
was aimed to determine nutritional contents of snail-eating turtle eggs for scientific approving the benefit for health.

2.

Materials and methods

2.1. Samples Collection

Snail-eating turtle (Malayemys macrocephala) eggs were collected from the turtles farm in Bangban district, Ayutthaya province. Sampling
eggs were 3-day-old and triplicate samplings were conducted. They were carefully handled in an ice-box, and transported to the laboratory,
kept in the refrigerator (@ 7C) before analyses.

2.2. Samples Preparation and Analyses for Chemical Compositions

Samples preparation and analyses for chemical compositions of amino acids, fatty acids and vitamins of the sampling eggs were
performed. Each sample was weighted and homogenized (by a laboratory blender, Moulinex A327, France), and subjected to proximate
analyses [8-10]. Profiles of amino acids were done by acid hydrolysis and derivertization with GC-FID detection [11], fatty acids profiles
by acid hydrolysis and methylation with Capillary GC detection [12]. Vitamin analysis was conducted by saponification and liquid
extraction of organic solvents with HPLC-Fluorescence detection [13]. All analyses were performed by a certified laboratory (ALS
Laboratory Group Co., Ltd., Bangkok).

* Corresponding author.
Email address: tkalayan@chula.ac.th

Tanasorn Tunsaringkarn et al.

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Agri culture Science De velopment s , 2(4) Apri l 2013

2.3. Minerals Analyses

All samples were oven dried before ashing at 550 C. for 5 hours. Addition of 20 mL of 1 N hydrochloric acid (HCl) followed by minerals
analysis by Inductively Couple Plasma (ICP) [14], to determine the mineral compositions of phosphorous (P), potassium (K), calciumiron
(Ca), magnesium (Mg), iron (Fe), manganese (Mn), copper (Cu), zinc (Zn) and nitrogen (N) analysis [15].
2.4. Samples Preparation for Sex Hormones Analyses
Egg samples were separated for egg white and egg yolk. and later homogenized. Each sample was weighed for 5.0 g, and 50.0 ml of
hexane was added and mixed for a 3-minute in a mixer. Solvent of the extract was evaporated in nitrogen gas. Sample extracts were soluble
in phosphate buffer (pH 7.0) and dimethylsulfoxide (DMSO) (1:1, vol/vol).Determinations of sex hormones of Luteinizing Hormone (LH),
Follicle Stimulating Hormone (FSH), Progesterone (P), Estradiol (E2), and Testosterone (T) were done by the Electrochemiluminescence
immunoassay (ECLIA) method [16] (Xin et al., 2010) using Roche Diagnostics kits. While, the determination of Prolactin (PRL) was done
by the Time-resolved fluoroimmunoassay (IFMA) method [17] (Diamandis, 1988) using the PerkinElmer Life and Analytical Sciences kit.
The hormonal analyses were performed by reference laboratory of King Chulalongkorn Memorial Hospital, Bangkok.
2.5. Statistical Analyses
Statistical analyses were carried out using descriptive statistical analysis and compared with One-Way ANOVA: Post Hoc Multiple
Comparison (LSD) between egg groups (whole eggs, egg whites and egg yolks). Mean differences with p<0.05, p<0.01and p<0.001 were
considered statistical significant. All statistical analyses performed by SPSS for window version 17.0.

3.

Results

Average proximate analyses of weight, ash, moisture, carbohydrate, protein, fat and total calories of the snail-eating turtle whole eggs were
11.55 g egg-1, 1.45 g 100g-1, 72.93 g 100g-1, 4.64 g 100g-1, 12.42 g 100g-1, 8.56 g 100g-1, 145.17 kcal 100g-1 respectively (Figure 1).
Most of parameters were higher in egg yolks than in the whole eggs and egg whites. There were significantly differences between each egg
group (p<0.001) except carbohydrate content, which e was not different in each group, and the moisture, which was the highest in egg
whites.
The essential amino acid (EAA) profiles, Young, 1994) of egg whites (227.55 g 100g-1) were slightly higher than the non-essential amino
acid (NEAA) profiles (216.16 g 100g-1) (Figure 2). Most of EAA were lysine (68.90 g 100g), leucine (53.83 g 100g-1 ) and phenylalanine
(32.37 g 100g-1) while most of NEAA were tyrosine (54.37 g 100g-1), glutamic acid (46.07 g 100g-1) and proline (31.27 g 100g-1).
The essential fatty acids (EFA) [1.33 mg 100g-1] contents of egg yolks were lower than the non-essential fatty acids (NEFA) [5.83 mg
100g-1] and the total fatty acids [of 7.87 mg 100g-1 }(Figure 3). Most NEFA of eggs yolk were oleic acid (2.65 mg 100g-1), palmitic acid
(1.24 mg 100g-1), palmitoleic acid (0.98 mg 100g-1). While, most EFA were arachidonic acid (0.47 mg 100g-1, linoleic acid (0.32 mg 100g1
) and linolenic acid (0.11 mg 100g-1).
The total unsaturated fatty acid (UFA) [4.85 mg 100g-1] was higher than saturated fatty acids (SFA, 2.08 mg 100g-1 ) with 2.33 folds (Figure
4). The UFA was composed of monounsaturated fatty acid (MUFA, 3.69 mg 100g-1) and polyunsaturated fatty acid (PUFA, 1.13 mg 100g1
).
In addition, the egg yolks had high contents of vitamin A (19.47 g 100g-1) and vitamin E (3.93 g 100g-1) (Figure 5).
The most total essential minerals of whole egg and egg yolk were nitrogen and phosphorous, while for the egg white they were nitrogen
and potassium. The most trace minerals of whole egg, egg white and egg yolk were all the same of zinc and iron (Figure 6 8).
For the sex hormones (LH, FSH, PRL, P, E2 and T), they exhibited similar patterns in whole egg and eggs yolk. The highest hormone of
whole egg and egg yolk was P (157.30 ng g-1 and 121.88 ng g-1 respectively), while the other male sex hormone was T (3.23 ng g-1 and 4.23
ng g-1 respectively) (Figure 9).

4.

Discussion

Carbohydrates are macronutrients which human needs to uptake in reasonably large quantities to be converted to energy for body needs
[18-19]. Dietary fiber has such a complicated chemical structure that the human body cannot metabolize it at all. So, fiber remains very
important for both health and weight control [20-21]. Moreover, it also helps to protect us against some serious diseases, including various
cancers [22]. The average carbohydrate include dietary fiber of snail-eating turtle egg was g 100g-1 and most of carbohydrate include
dietary fiber was in egg whites.

Determination of Chemical Compositions of Snail-Eating Turtle (Malayemys macrocephala) Eggs

33

Agri culture Scienc e Developments , 2(4) April 2013

Lysine plays many significant roles in the human body. It is crucial for the proper absorption of calcium from the gastrointestinal tract and
helps in conserving calcium [23]. It facilitates the production of enzymes, hormones and antibodies and assists in the formation of muscle
protein. Lysine aids in the synthesis of collagen, an important constituent of bones and connective tissues. Besides this, L-lysine benefits
involve stimulating the production of creatinine, which is responsible for converting fatty acids into energy. Leucine assists to regulate
blood sugar and energy levels, production of the human growth hormone, wound healing as well as the growth and repair of muscle tissue.
Moreover, it can also assist in the prevention of the breakdown of muscle proteins that may occur after severe stress or trauma [24-25]. For
tyrosine is benefit for depression treatment and improved mental clarity, appetite suppression, improved endurance, more energy, improved
thyroid function, decreased pre-menstruation symptoms (PMS), male sex drive and emotional health [26-28]. A major deficiency in
tyrosine may result in hypothyroidism - an enlargement of the thyroid gland (goiter), severe growth failure, and retardation of central
nervous system development [29]. Both EAA and NEAA are health benefits for solving malnutrition problem and improve the quality of
life worldwide [30-31].
Arachidonic acid is a polyunsaturated fatty acid present in the phospholipids which abundant in the brain, muscles, liver. It is necessary for
the repair and growth of skeletal muscle tissue, early neurological development and disorders [32-34]. While, oleic acid reduce the
cardiovascular risk by reducing blood lipids for long chain omega-3 PUFA (eicosapentaenoic acid-EPA and docosahexaenoic acid-DHA)
[35].
Unsaturated to saturated fatty acid ratios was 2.33 which EFA are omega-3 (-3, linolenic acid) and omega-6 (-6, linoleic acids) which
humans cannot synthesize, and must be obtained through diet. Linolenic acid (ALA) will convert into eicosapentaenoic acid (EPA), and
later into docosahexaenoic acid (DHA). EPA and the GLA synthesized from linoleic acid are later converted into hormone-like compounds
known as eicosanoids, which aid in many bodily functions including vital organ function and intracellular activity. The level of -3 in eggs
can be naturally high advantage, particularly through their DHA content may be similar to those produced following the fish oils
consumption and their ability can decrease risk factors for heart disease [36]. The previous conducted that a chicken egg consumption a day
is unlikely to have impact on cardiovascular disease risk in non diabetes and egg consumption has been promoted satiety [37]. The other
study showed that eggs make a significant contribution to the anti-inflammatory effects of carbohydrate restricted diets (CRD), possibly
due to the presence of cholesterol, which increases HDL-C and to the antioxidant lutein which modulates certain inflammatory responses
[38]. Thus, they can support the cardiovascular, reproductive, immune, and nervous systems [39-41]. The snail-eating turtle egg is the high
of -3 to -6 ratio at 0.23, and which is the better value for human nutritional need [42]. The more increasing -3 to -6 ratio, the more
benefit for health.

Trans fat is the UFA with trans-isomer fatty acids which may be MUFA or PUFA. The primary health risk identified for trans fat
consumption is an elevated risk of coronary heart disease [43]. This result showed that trans fatty acids was 0.04 g 100g-1 (Figure 4) which
it was much less than cis-isomer. The previous studies reported that dietary sources of maternal diet affected on fatty acid compositions
[44-45] and lipid compositions in egg yolk are important for viability, growth and/or embryo development [46-48]. Food manufacturers
have used it less because of concerns over the health effects of trans fat. Food manufacturers in the United States and many other countries
list the trans fat content on nutrition labels. They recommended limit less than 0.5 grams of trans fat per serving.
It was inconsistent with previous report that egg yolk is one of the few foods naturally containing vitamin D [49]. The retinol level in this
study was higher than recent study (0.23 g g-1 yolk) in Japan [50]. Vitamin D is a fat-soluble vitamin that is naturally present in very few
foods and essential for promoting calcium absorption in the gut and maintaining adequate serum calcium and phosphate concentrations to
enable normal mineralization of bone and prevent hypocalcemic tetany, also helps protect older adults from osteoporosis. Vitamin D has
other roles in human health, including modulation of neuromuscular and immune function and reduction of inflammation [51-52] but it was
low level in this eggs yolk. Vitamin A promotes growth with healthy teeth, bones, soft tissue, mucus membranes, and skin, the immune
system, reproduction, and vision [53]. Vitamin A deficiency is a public health problem in more than half of all countries, especially in
Africa and South-East Asia, hitting hardest young children and pregnant women in low-income countries [54]. A vitamin E is an
antioxidant also known as tocopherol. It plays a role in the formation of red blood cells, helps the body use vitamin K and prevent chronic
diseases of heart disease, cancer, neurodegenerative diseases such as dementia, parkinsons disease and amyotrophic lateral sclerosis
(ALS) [55]. There are many reports of health benefits from vitamin E diets [56-57]. However, be very careful about taking large amounts of
fat-soluble vitamin supplements of vitamins A, D, E, and K. Because these vitamins are stored in fat cells, they can build up in your body
and may cause harmful effects. The Recommended Dietary Allowances (RDAs) for vitamins reflect how much of each vitamin most
people should get each day and used as goals for each person. Snail-eating turtle eggs yolks had high contents of vitamin A which may be
relevant to the successful development of embryo [58]. Vitamin E is very effective natural antioxidant and may have important role in
preventing peroxidative damage to developing tissues [59]. Thus, snail-eating turtle may resist the embryo development. The body requires
vitamins to support its basic biochemical functions, and deficiencies over time can lead to illness and disease.
Nitrogen functions as the component of nucleic acids, proteins, hormones, coenzymes [60]. Phosphorus is an important constituent of
adenosine triphosphate (ATP) and nucleic acid and is also essential for acid-base balance, bone and tooth formation [61]. Red blood cells
can not function properly without iron in haemoglobin, the oxygen-carrying pigment of red blood cells. Iron is also an important
component of the cytochromes that function in cellular respiration [62]. The enrichment of iron in egg would provide improving the
nutrition status of people especially in the risk of iron deficiency or anemia group especially infant, children, pregnant women and
socioeconomic groups [63-68]. Zinc functions as a cofactor and is a constituent of many enzymes like lactate dehydrogenase, alcohol
dehydrogenase, glutamic dehydrogenase, alkaline phosphatase etc., and zinc dependent enzymes are involved in macronutrient metabolism

Tanasorn Tunsaringkarn et al.

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Agri culture Science De velopment s , 2(4) Apri l 2013

and cell replication [69-70]. Moreover, zinc and iron are important co-factors found in the structure of certain enzymes and are
indispensable in numerous biochemical pathways. , the enrichment of zinc would be benefit for reduction of diarrhea and pneumonia
mortality in children [71-72]. Whole of snail-eating turtle eggs, nitrogen was both high in egg whites and yolks while iron was higher in
egg whites but zinc was higher in egg yolks. The mineral elements which contribute to variations in the degree of physiological response to
deficient or toxic levels. These relationships make it difficult to determine the optimum dietary level for the individual elements required
for humans. Micronutrients such as vitamins and minerals are important for health. One out of three people in developing countries are
affected by deficiency such as vitamin A and iron [73].
Health benefits of natural P are antidepressant, balancing blood sugar level, decreasing PMS and menopause symptoms and weight loss
[74]. The PRL of eggs was very low level which human PRL functions during pregnancy and lactation [75-76]. A poor life can be caused
by hormonal problems. Steroid hormones varied with the species reproductive strategy and social requirement which may affect male and
female offspring differently [77]. Moreover, sex steroid of maternal origin could be found in egg yolks of many species [78-79] and
correlated with maternal circulating plasma [80]. Snail-eating turtle eggs should be more benefit for women health.

Figure 1. Proximate analyses of snail-eating turtle whole eggs

Esse ntial Amino Acid (EAA)

Figure 2. Amino Acids of snail-eating turtle egg whites

s
AA
lE

al
in
e

0
V

ni
sp
n
ar
tic e
A
ci
d
C
G
ys
lu
tin
ta
e
m
ic
A
ci
d
G
ly
cin
e
H
is
ti d
in
e
Pr
ol
in
e
Se
ri
ne
Ty
ro
To
sin
ta
e
lN
EA
A
s

rg
i

la
ni
ne

50

To
ta

50

100

ne

100

150

Ly
si
ne
M
et
hi
on
in
Ph
e
en
yl
al
an
in
e
Th
re
on
in
e
Tr
yp
to
ph
an

150

200

Le
uc
i

200

250

Is
ol
eu
cin
e

Concentration (mg 100g-1 )

250

Concentration (mg 100g-1 )

Non-Esse ntial Amino Acid (NEAA)

Determination of Chemical Compositions of Snail-Eating Turtle (Malayemys macrocephala) Eggs

35

Agri culture Scienc e Developments , 2(4) April 2013

0.5
0.45
0.4
0.35
0.3
0.25
0.2
0.15
0.1
0.05
0

Ei
co

g-

Figure 3. Fatty acid profiles of snail-eating turtle egg yolks

Total Fatty Acid

6
Concentration

5
4
3
2
1

(m

/S
FA
FA

Tr
an

sF

00
g
g1

g1
(m

FA
U

-1
)

-1
)
00
g

-1
)
00
g
g1

SF

(m

g1
(m

PU
FA

UF
A

(m

g1

00
g

00
g

-1
)

-1
)

Figure 4. Total fatty of snail-eating turtle egg yolks

Vitamin
25
20
15
10
5
0
Vitamin A

Vitamin D

Vitamin E

Figure 5. Vitamin A, D and E of snail-eating turtle egg yolks

-6
3/
-

ac
id
di
en
oic
a
sa
ci d
t ri
en
oi
Ei
ca
co
sa
c id
tr
ien
oic
Ar
ac
ac
id
hi
Ei
do
co
n
i
sa
ca
pe
c id
nt
ae
Do
no
co
ic
sa
ac
he
id
xa
en
oi
ca
c id

ni
c

Ei
co

Li
no
le

Li
no
le

ic
a

cid

Essential Fatty Acid (EFA)

sa

Concentration (mg 100 g -1 )

3
2.5
2
1.5
1
0.5
0

Concrentration (ug 100g-1 )

Concentration (mg 100g-1)

Non-Essential Fatty Acid (NEFA)

Tanasorn Tunsaringkarn et al.

36

Agri culture Science De velopment s , 2(4) Apri l 2013

Trace Mineral

ce
Zi
n

op
pe
r
C

M
an

ga
ne
se

Ir
on

um

80
70
60
50
40
30
20
10
0

M
ag
ne
si

al
ciu
m
C

Po
ta
ss
iu
m

Ph
os
ph
or
o

itr
N

us

Concentration (mg L-1 )

10
9
8
7
6
5
4
3
2
1
0
og
en

Concentration (%)

Esse ntial Mine ral

Figure 6. Minerals of snail-eating turtle whole eggs

Trace Mine ral

9
8
7
6
5
4
3
2
1
0

Concentration (mg g -1 )

Concentration (%)

Esse ntial Mine ral

it

n
ge
ro

ou
or
h
p
os
Ph

m
siu
as
t
Po

ciu
al

70
60
50
40
30
20
10
0
on
Ir

m
iu
es
n
ag
M

r
pe
op

e
es
an
g
an
M

e
nc
Zi

Figure 7. Minerals of snail-eating turtle egg whites

Trace Mine ral

10
9
8
7
6
5
4
3
2
1
0

Concentration (mg L-1 )

Concentration (%)

Esse ntial Mine ral

itr

en
og
P

us
ro
o
h
sp
ho

m
siu
as
t
Po

m
ciu
al

um
si
ne
g
a
M

80
70
60
50
40
30
20
10
0
on
Ir
a
M

e
es
an
g
n

Figure 8. Minerals of snail-eating turtle egg yolk

r
pe
op

e
nc
Zi

Determination of Chemical Compositions of Snail-Eating Turtle (Malayemys macrocephala) Eggs

37

Agri culture Scienc e Developments , 2(4) April 2013

Hormone

Hormone

180

140

160

120

140
C o nc entra t io n

Concentration

100
80
60
40

120
100
80
60
40

20

20
0

0
LH (IU g-1) FSH (IU g-1) PRL (ng g-1)

P (ng g-1)

E2 (ng g-1)

A) Egg yolk

T (ng g-1)

LH (IU g-1)

FSH (IU g-1)

PRL (ng g-1)

P (ng g-1)

E2 (ng g-1)

T (ng g-1)

B) Whole egg

Figure 9. Sex hormones of snail-eating turtle egg yolk and whole egg

5.

Conclusion

Our results revealed that snail-eating turtle eggs had the potential good nutrients of carbohydrate, fat, protein, mineral and hormone. It also
had essential amino acid of lysine and leucine, fatty acid of arachidonic acid (-6), linoleic acid (-3). Snail-eating turtle eggs may be an
alternative source of vitamin A, hormone progesterone (P), nitrogen, iron and zinc. Although, there were nutritional differences between
egg yolk and egg white in which more nutrients would be found in the egg yolk.

Acknowledgement
This work was funded and supported by Surveillance Center on Health and Public Health Problem, College of Public Health Sciences,
Chulalongkorn University Centenary Academic Development Project, Chulalongkorn University.

Notes
The authors declare no competing financial interest.

Abbreviation
AA, amino acid; EFA, essential fatty acid; NEFA, non-essential fatty acid; MUFA, monounsaturated fatty acid; PUFA, polyunsaturated
fatty acid; SFA; saturated fatty acid; UFA, unsaturated fatty acid; LH, luteinizing hormone; FSH, follicle stimulating hormone; PRL,
prolactin; P, progesterone; E2, estradiol; T, testosterone; DMSO, dimethylsulfoxide; IFMA, time-resolved fluoroimmunoassay; ECLIA,
electrochemiluminescence immunoassay; ALA, linolenic acid; EPA, eicosapentaenoic acid; DHA, docosahexaenoic acid; GLA, gammalinolenic acid.

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