jurnal farmasi bahari te

1. International Journal of Agricultural Policy and Research Vol.2 (11), pp. 373-378, November 2014
Available online at http://www.journalissues.org/IJAPR/ http://dx.doi.org/10.15739/IJAPR.009
© 2014 Journal Issues ISSN 2350-1561
Original Research Article
Phytochemical constituents and bioscreening activities of
green algae (Ulva Lactuca )
Dalia F. Abd Elmegeed,
Doaa A. Ghareeb*,
Muhammed Elsayed
and
Muhammad El-Saadani
Biochemistry department,
Faculty of Science,
Alexandria University,
Alexandria, Egypt.
*Corresponding Author Email:
d.ghareeb@yahoo.com Tel.:
+201277291144
Accepted 30 August, 2014
This study was carried out to investigate the phytochemical and biological
screening of green algae (Ulva lactuca, UL) crude extract and polysaccharide
(PS), U. lactuca was collected from Abu Kir Beach, Alexandria coast, Egypt.
Phytochemical screening was assessed by chemical analysis, while the
coagulatory effect was assessed by measuring prothrombin time (PT) and
activated partial thromboplastin time (APTT). In addition, antioxidant
property was measured in form of reduction in TBRSA formation in liver
homogenate. Finally, the safety of extract was assessed by RBC toxicity test.
Our results showed that, the green algae extract contained alkaloids,
flavonoids and saponins that were 79.58, 2.36 and 0.03 mg/100 g algae dry
weight, respectively, and the concentration of total phenols was 14.223
mg/100g dry algae tissue. The tested extracts showed antioxidant properties
as it inhibited the induction of lipid peroxidation. The U. lactuca crude
extract showed anticoagulation property while PS showed coagulation
behavior. Finally, UL crude extract has non hemolytic effect on the red blood
cells while PS solution has semi hemolytic effect on red blood cells. In
conclusion, the UL extracts as natural product have high antioxidant activity
due to the presence of total phenolic compounds and the coagulatory effect
[assessed by measuring prothrombin time (PT) and activated partial
thromboplastin time (APTT)] may be beneficial in the treatment of some
cardiovascular diseases.
Key words: Ulva lactuca, TBARS, antioxidant, coagulation, anti-inflammatory and
RBC toxicity
Abbreviations: APTT, Activated partial thromboplastin time; BHT, butylated
hydroxytoluen; PS, polysaccharide solution; PT, prothrombin time; TBA,
thiobarbituric acid; TBARS, thiobarbituric acid reactive substances; TCA,
trichloroacetic acid; UL, Ulva lactuca.
INTRODUCTION
Nature has provided many things for humankind over the
years, including the tools for the first attempts at
therapeutic intervention (Nakanishi, 1999). The term
natural products is quite commonly understood to refer to
herbs, herbal concoctions, dietary supplements, traditional
Chinese medicine, or alternative medicine (Holt and Chandra,
2002). However, generally, natural product is a chemical
compound or substances produced by a living

2. Int. J. Agric. Pol. Res. 374
organism such as prebiotic origin or originate from
microbes, plants, or animal sources (Nakanishi, 1999). As
chemicals, natural products include such classes of
compounds as terpenoids, polyketides, amino acids,
peptides, proteins, carbohydrates, lipids, nucleic acid bases,
ribonucleic acid (RNA), desoxyribonucleic acid (DNA)
among many others (Shayne, 2005). It usually possess
pharmacological or biological activity suitable for use in
pharmaceutical drug discovery and drug design. A natural
product can be considered as such even if it can be
prepared by total synthesis (Newman and Cragg, 2007).
Recent reports indicate that there is an inverse
relationship between the dietary intake of antioxidant-rich
foods and the incidence of human diseases. As a result,
many researchers have focused on natural antioxidants as
numerous crude extracts and pure natural compounds have
previously been reported to exhibit/possess antioxidant
properties in the plant kingdom (Halliwell, 1997).
Recently, aquatic habitats have increasingly been shown
to provide a rich source of natural bioactive compounds
with hypocholesterolemia, antinflammatory, antiviral,
antineoplastic, antimicrobial and hypertensive properties.
According to their chemical structure, most isolated
compounds belong to sulfated polysaccharides, phenolics,
terpenoids, lactons, sterol and fatty acids (McDermid and
Stuercke, 2003; Qi et al., 2005; Duan et al., 2006).
The genus Ulva popularly known as sea-lettuce is one of
the most common and abundant green macroalgae
throughout the world (Lahaye and Robic, 2007). Despite its
wide distribution, it is poorly utilized (Ray and Lahaye,
1995) and only a small part of its biomass is used as food or
animal feed due to its nutritional components (vitamins,
oligoelements, minerals and dietary fibers) (Pengzhan et al.,
2003); organic crop fertilizer (Mulbry et al., 2005), effluent
biofilter (Msuya and Neori, 2002) and more recently, as
plant protectant (Cluzet et al., 2004).
Some seaweed contains high amounts of essential
proteins, vitamins, and minerals (Norziah and Ching, 2000).
Velichko and Shevchenko (1998) have reported that
seaweeds as a dietary supplement are found to be good for
the prophylaxis of coronary atherosclerosis. Its antioxidant
activity is one of the most important and active in marine
bioactive substances and lots of algal and algae-derived
compounds such as carotenoids, phenolics, terpenoids and
sulphated polysaccharides exhibit potent antioxidant
activities. The antioxidant activities of these compounds are
mainly attributed to scavenging activity against superoxide
and hydroxyl radicals, chelating ability, quenching singlet
and triplet oxygen and reducing power (Ruberto et al., 2001
and Athukorala et al., 2006).
One of the products easily obtainable from Ulvales
seaweeds is a polysaccharide, a phycocolloid (Percival,
1979) commonly known as ulvan. Studying the saccharidic
composition of ulvan, Percival and Wold (1963) showed
that by this name is indicated a family of water-soluble
anionic polysaccharides structurally heterogeneous
containing galactose, xylose, rhamnose, glucuronic acid and
iduronic acid arranged in an essentially linear fashion
(Percival and Wold, 1963). The composition and extent of
these structural motifs, called aldobiuronic A3S and B3S
sequences, can vary depending on the sources and on the
year/period of collection of the algal material.
Algal polysaccharides have been demonstrated to play an
important role as free-radical scavengers in vitro and
antioxidants in the prevention of oxidative damage in living
organisms (Zhang et al., 2004). Furthermore, many
polysaccharides found in seaweeds have diverse biological
activities, including effects on the immune system and
cancer (Yashizawa et al., 1995).
Altogether, this study aimed to investigate the biological
activities of the methanolic crude extract and
polysaccharide extract of green algae (Ulva lactuca) that
was collected from Abu Kir Beach, Alexandria Coast.
MATERIALS AND METHODS
Trichloroacetic acid (TCA), thiobarbituric acid (TBA) and
butylated hydroxytoluen (BHT) were purchased from
Sigma Chemical Co. (St. Louis, Mo, USA). Kits for PT and
APTT were purchased from Futura System (Rome, Italy).
All other chemicals and reagents were of the highest quality
available commercially.
Green algae (U. lactuca) were collected from Abu Kir
Beach, Alexandria Coast, and identified by Prof. Dr. Samy
Shaalan, Microbiology and Botany Department, Faculty of
Science, Alexandria University, Egypt.
Preparation of U. lactuca extracts
Polysaccharide extraction was performed according to
Hwang et al. (2008). The algae was washed under tap water
and air dried. The polysaccharide (PS) was firstly isolated
by steeping dried UL powder (40 g) in 1 L of distilled water
for 3 h at 80°C. The aqueous extract was then clarified by
centrifugation at 10,000 rpm at 4°C for 20 min and
subsequent filtration through filter paper No. 3 to remove
insoluble materials. The PS in the aqueous extract was
precipitated with two volumes of ethanol and recovered by
filtration. The PS recovered precipitate (6.4 g) was used as
the crude UL –PS preparation.
Secondly, UL methanolic crude extract was prepared by
soaking dried powdered UL in methanol overnight (1:50,
w/v) at room temperature. The UL methanolic extract was
then collected by water pump vacuum filtration, condensed
by a rotary evaporator (Buchi, Switzerland) at 50°C (Taskin
et al., 2007) and lyophilized to obtain powdered UL crude
extract. Each powder was dissolved in 1% saline (NaCl, 0.9
%) which was used for further investigations (tested
extract).
The phytochemical screening and quantitative estimation
of the concentration of chemical constituents were carried

3. Table 1. Qualitative phytochemical screening of
the green algae Ulva lactuca.
Test Green algae
Alkaloids +
Tannins _
Phlobatannins _
Saponins +
Flavonoids +
Terpenoids +
Cardiac Glycosides +
+ indicates the presence of the component.
- indicates the complete absence of the component
out according to Edeoga et al. (2005).
Phenol sulfuric acid test for PS solution and total phenolic
content of UL solution were carried out according to Liu et
al. (1973) and Zahra et al. (2007), respectively.
Determination of biological activities
Effect on coagulation pathway was done by assessing the
prothrombin time (PT) and activate partial thromboplastin
time (APTT) activity according to Athukorala et al. (2007).
50 µl of each tested fraction was added to 100 µl human
plasma (blood withdrawal was carried out from healthy
subjects after approval was sought and signed by subjects
under the supervision of Dr. Doaa Ahmad Ghareeb, clinical
biochemist, according to the Alexandria University Ethics’
Regulation Book). The mixture was incubated for 5 min at
37°C as described by Athukorala et al. (2007).
The toxicity study was estimated by the hemolysis test
which was performed according to the American Society for
Testing and Materials (ASTM, 2000) (ASTM F 756-00).
Different concentrations of PS or UL crude extract (0.25,
0.5, 0.75 and 1 mg) were placed in test tubes and mixed
with 7 ml of phosphate buffer saline. 100 μl of blood was
mixed with each concentration and maintained at 37°C for
3 h. Positive and negative controls were prepared by adding
the same amount of blood to 7 ml of water and PBS,
respectively. Each tube was gently inverted twice, each for
30 min to maintain contact between blood and the material.
After incubation, each fluid was transferred to a suitable
tube and centrifuged at 2000 rpm for 15 min. The
hemoglobin released by hemolysis was measured by the
optical densities (OD) of the supernatants at 540 nm using a
spectrophotometer. The percentage of hemolysis was
calculated as follows:
Hemolysis (%) = (OD sample − OD negative control) / (OD
positive control – OD negative control) * 100
According to ASTM (2000) materials can be classified into
three different categories according to their hemolytic
Elmegeed et al. 375
indices (hemolysis %). Materials with percentages of
hemolysis of over 5% are considered hemolytic while an
index between 5 and 2% are classified as slightly hemolytic.
Finally, when the material presents a hemolysis percentage
below 2%, it is considered as a non-hemolytic material.
Antioxidants properties of fraction were done by
determination of liver homogenate with thiobarbituric acid
reactive substances (TBARS) scavenging according to
Tappel and Zalkin (1959). 2 ml of each tested fraction was
incubated for 45 min at 37°C. 100.25 μL of FeSO4.7H2O
(0.02 M) and 10 μL of H2O2 (0.4 M) were added to each
sample and incubated for 30 min at 37°C. Finally, 80.6 μL of
1% BHT were added to the previous mixture. All samples
were cooled and centrifuged at 3000 rpm for 15 min. 1 mL
of the supernatant of each sample was added to 1 mL of
TCA (15 %) to precipitate protein, and the samples were
re-centrifuged at 3000 rpm for 10 min. To each 1 ml of
supernatant, 0.5 ml of TBA (0.7 %) was added and the
mixture boiled in a boiling water bath for 45 min. The
absorbances, using a M108 Programmable Visible Range
spectrophotometer (Camspec Analytical Instruments
Ltd.UK), of samples were read at 532 nm against a blank
reagent.
Statistical analyses
Data were analyzed by one-way analysis of variance
(ANOVA) using Primer of Biostatistics (Version 5) software
program. Significance of means ± SD (standard deviation)
was determined in groups by the multiple comparisons
Student-Newman-keuls test (Khaltree and Naik, 2006).
RESULTS
Qualitative phytochemical screening of the powdered green
algae reveals the presence of alkaloids, flavonoids,
saponins, terpenoids and cardiac glycosides. However,
tannins, phlobatannins and steroids were not detected
(Table 1). The amounts of alkaloids, flavonoids and
saponins are 79.58, 2.36 and 0.03 mg/100g algae dry
weight, respectively, and the concentration of total phenols
is 14.223 mg/100g of dry algae tissue (Table 2).
Our results showed that the concentration of phenol
sulfur in polysaccharide extract of green algae is 204.309
mg/g crude extract. While the concentration of total
phenolic content in UL crude extract is 1.6 µg/g crude
extract as shown in (Table 3).
On one hand, UL crude extract exhibited anticoagulation
properties as PT and APTT were increased than in the
control (Table 4). On the other hand, the PS solution
exhibited coagulation property as it decreased both the PT
and APTT than in the control at p<0.001.
The comparative quantitative estimation of RBC s toxicity
indicated that all tested concentrations of UL crude extract
showed no hemolytic effect on the red blood cells while

4. Int. J. Agric. Pol. Res. 376
Table 2. Quantitative phytochemical screening of the green
algae Ulva lactuca
Test Concentration*
Alkaloids 79.58 ± 4.5
Saponins 0.03 ±0.0002
Flavonoids 2.36 ± 0.04
Total phenols (mg/ml) 14.223 ± 1.2
* Values are expressed as mg/100g of dry algae weight.
Each value is the mean ±SD of triple determinations.
Table 3. Phenol sulfur acid test of PS solution and total phenolic content of UL crude extract
Green algae extracts Phenol sulfur concentration (mg/g Total phenolic content concentration
crude extract) (µg/g crude extract)
PS solution 204.309 ± 26.2 ------
UL crude extract ------- 0.016 ± 0.001
Each value is the mean ±SD of triple determinations.
Table 4. Effect of different algae extracts on blood coagulation tests
Algae extract PT (sec) APPT (sec)
Control 45.500 ± 4.850 41.000 ± 4.240
PS solution 25 ± 5.568 * 24 ± 2*
UL crude extract 92.000 ± 4.14* 62.000 ± 6.970*
*: Means significantly different at the level of p<0.001.
Each value is the mean ±SD of triple determinations.
Table 5. Effect of PS solution and UL crude extract on RBC s toxicity
Concentration (mg) UL crude extract PS solution
Haemolysis%
0.25 1.46 6.2
0.5 0 5
0.75 -1.6 3.5
1 -0.65 2.6
Each value is the mean ±SD of triple determinations.
only 0.25 mg concentration of PS solution showed
hemolytic effect with the other concentrations showing a
semi hemolytic effect on red blood cells as shown in Table
5. The tested extracts showed antioxidant properties and
inhibited the induction of lipid peroxidation by decreasing
the level of induced TBARS in the liver homogenate as
shown in Table 6.
DISCUSSION
The algal extracts were examined to estimate their effect on
the antioxidant status in liver homogenate. Oxidative stress
occurs as a result of increased production of destructive
reactive oxygen species (ROS) (including oxygen ions, free
radicals and peroxides) more than the body scavenger
system consisting of enzymatic and non-enzymatic
antioxidants (Leeuwenburgh et al. 2001).
The most famous naturally occurring compounds with
antioxidant capacity are alkaloids, flavonoids and total
phenols found in foods such as onion and garlic which show
functional health benefits in the reduction of cardiovascular
disease risk by lowering serum cholesterol and blood
pressure (Banerjee and Maulik, 2002). They have
anticarcinogenic, antidiabetic, antiplatelate aggregation and
antibiotic effects (Augusti, 1996; Lau, 1998; El-Demerdash
et al., 2005). Referring to our data, algal extracts contain a
high amount of flavonoids and total phenolic contents that

5. Table 6. Effect of PS solution and UL crude
extract on lipid peroxidation
Algae % of inhibition
PS solution 44.16 ± 1.2
UL crude extract 56.29 ± 0.9
Each value is the mean ±SD of triple determinations.
gives them a high antioxidant capacity enabling them to be
used for the treatment of several diseases (Pourmorad et
al., 2006). Furthermore, our algal extracts showed powerful
inhibitory action against oxidative stress in liver tissue.
Thus, these extracts could be used in the treatment of
oxidative stress- associated liver damage as Qi et al. (2006)
and Sire et al. (1983) have reported that U. lactuca
polysaccharides have the ability to protect the liver against
free radicals.
A blood clot is developed due to the failure of hemostasis
which causes vascular blockage and sometimes leading to
death. Natural plants also possess anticoagulant property
which is regarded as safe compared to other synthetic
agents (Erasto et al., 2007). It is reported that total phenolic
compound present in plants possesses both antioxidants
and anticoagulation properties (Anitha and Ponbavani,
2013). In agreement with this report, our data showed the
same results as UL crude extract has anticoagulation
property which could be due to the presence of total
phenolic compounds. Anticoagulant agents, as detected in
our crude extract, are compounds with an ability to prevent
the activation of the coagulation cascade and may be used
as a treatment for stroke and blood coagulation diseases
(Savelieva, 2007). Therefore, the crude extract could be
used as a new candidate in the treatment of blood
coagulation diseases. On the other hand, the PS solution
showed coagulation effect. It is reported that
polysaccharides isolated from plants activate the blood
coagulation system (Glu, 1986).
Haemolysis is regarded as an especially significant
screening test as it provides quantification of small levels of
plasma haemoglobin which may not be measurable under
in vivo conditions. As reported in literature (ISO 10993 –4,
1999), it is not possible to define a universal level of
acceptable or unacceptable amounts of haemolysis.
Although, by definition, a blood-compatible material should
be non-haemolytic, in practice several medical devices
cause haemolysis. This means that when such haemolytic
effect takes place, it is important to make sure that clinical
benefits outweigh the risks and that the values of
haemolysis are within acceptable limits. Our results
indicated that all tested concentrations of UL crude extract
have non hemolytic effect on red blood cells while only 0.25
mg concentration of PS solution has hemolytic effect
however, the other concentrations showed semi hemolytic
effect. In conclusion, the green algae could be used as
Elmegeed et al. 377
therapeutic agent since it contains compounds that have
potent antioxidant activity hence may be used as a
treatment for blood coagulation disorders.
REFERENCES
American Society for Testing and Materials 2000. ASTM F
756-00: Standard Practices for Assessment of Haemolytic
Properties of Materials, Philadelphia.
Athukorala Y, Lee K W, Kim S K, Jeon Y J (2007)
.Anticoagulant activity of marine green and brown algae
collected from Jeju Island in Korea. Biore-source
Technology, 98: 1711-1716.
Augusti KT (1996). Therapeutic values of onion (Allium
cepa L.) and garlic (Allium sativum L.). Indian J. Biol., 34:
634-640
Banerjee SK, Maulik SK (2002) .Effect of garlic on
cardiovascular disorders. Nutr. J., 4:1-14.
Cluzet S, Torregrossa C,Jacquet C, Lafitte C, Fournier J,
Mercier L, Salamagne S, Briand X, Esquerre-Tugaye M T,
Dumas B (2004). Gene expression profiling and
protection of Medicago truncatula against a fungal
infection in response to an elicitor from green algae Ulva
spp. Plant Cell and Environ., 27: 917-928.
Duan XJ, Zhang WW, Li XM, Wang BG (2006). Evaluation of
antioxidant property of extract and fractions obtained
from a red alga, Polysiphonia urceolata. Food Chem., 95:
37-43.
Edeoga HO, Okwu DE, Mbaebie BO (2005). Phytochemical
constituents of some Nigerian medicinal Plant. African J.
Biotechnology, 4(7): 685- 688.
El-Demerdash FM, Yousef MI, Abou El-Naga NI (2005).
Biochemical study on the hypoglycemic effects of onion
and garlic in alloxan-induced diabetic rats. Food Chem.
Toxicol., 43: 57-63.
Erasto P, Grierson D, Afolayan A (2007). Antioxidant
constituents in Vernonia amygdalina leaves.
Pharmaceutical Biology. 45: 195-199.
Glu L (1986). Effect of plant polysaccharides on blood
coagulation activity in animals. Farmakologiia i
Toksikologiia. 49(4): 38-40]
Halliwell B (1997). Advances in pharmacology. Academic
Press, 38: 3-17.
Holt GA, Chandra A (2002). Herbs in the modern healthcare
environment— an overview of uses, legalities, and the
role of the healthcare professional. Clin. Res. Regulatory
Affairs (USA), 19: 83–107.
Hwang HJ, Kwon MJ, Kim IH, Nam TJ (2008). The effect of
polysaccharide extracted from the marine alga
Capsosiphon fulvescens on ethanol administration. Food
Chem. Toxicol., 46: 2653–2657.
International Standard Organization (1999). ISO 10993-4:
Biological Evaluation of Medical Devices – Part 4 –
Selection of Tests for Interaction with Blood, Geneva.
Khaltree R, Naik S (2006). Applied multivariate statistics

6. Int. J. Agric. Pol. Res. 378
with SAS software. (2nd ed.). John Wiley and Sons Co. pp:
1-340.
Lahaye M, Robic A (2007). Structure and functional
properties of ulvan, a polysaccharide from green
seaweeds. Biomacromolecules, 8:1765-1774.
Lau BHS (1998). Detoxifying radio-protective and
phagocyte-enhancing effects of garlic. International and
Nutrition Reviews, 9: 27-31.
Leeuwenburgh C, Heinecke J (2001) .Oxidative stress and
antioxidants in exercise. Current Med. Chem., 8: 829-838.
Liu D, Wong PTS, Dutka BJ (1973). Determination of
carbohydrates in lake sediment by a modified
phenolsulfuric acid method. Water Res., 7: 741-746.
McDermid KJ, Stuercke B (2003). Nutritional composition of
edible Hawaiian seaweeds. J. Appl. Phycol., 15: 513-524.
Msuya FE, Neori A (2002). Ulva reticulata and Gracilaria
crassa: macroalgae that can biofilter effluent from
tidalfishponds in Tanzania. West. Indian Ocean J. Mar. Sci.,
1: 117-126.
Mulbry W, Westhead K, Pizarro C, Sikora L (2005).
Recycling of manure nutrients: use of algal biomass from
dairy manure treatment as a slow release fertilizer.
Bioresource Technol., 96:451-458.
Nakanishi K (1999). An historical perspective of natural
products chemistry. In S. Ushio (Ed.), Comprehensive
Natural Products Chemistry. Elsevier Science B.V.,
Amsterdam, 1: 23–40.
Newman DJ, Cragg GM (2007). Natural products as sources
of new drugs over the last 25 years. J. Nat. Products,
70:461-477.
Norziah MH, Ching CY (2000). Nutitional composition of
edible seaweed Gracilaria changgi. Food Chemistry, 68:
69-75.
Pengzhan Y, Quanbin Z, Ning L, Zuhong X, Yanmei W, Zhi’en
L. (2003). Polysaccharides from Ulva pertusa
(Chlorophyta) and preliminary studies on their
antihyperlipidemia activity. J. Appl. Phycol., 15:21-27.
Percival E (1979).The polysaccharides of green, red and
brown seaweeds: their basic structure, biosynthesis and
function. Br. Phycol J., 14: 103-117.
Percival E, Wold JK (1963). The acid polysaccharide from
the green seaweed Ulva lactuca. Part II. The site of the
ester sulphate. J. Chem. Soc. 5459-5468.
Pourmorad F, Hosseinimehr J, Shahabimajd N
(2006).Antioxidant activity, phenol and flavonoids
contents of some selected Iranian medicinal plants. Afr. J.
Biochem., 5: 1142-1145.
Qi H, Zhang Q, Zhao T, Hu R, Zhang K, Li Z. (2006). In vitro
antioxidant activity of acetylated and benzoylated
derivatives of polysaccharide extracted from Ulva
pertusa (Chlorophyta). Bioorg. Med. Chem. Lett. 16:2441-
2445.
Qi H, Zhao T, Zhang Q, Li Z, Zhao Z , Xing R (2005).
Antioxidant activity of different molecular weight
sulfated polysaccharides from Ulva pertusa Kjellm
(Chlorophyta). Appl. Phycol., 17: 527-534.
Ray B, Lahaye M (1995). Cell-wall polysaccharides from the
marine green-alga Ulva-rigida (Ulvales, Chlorophyta) -
chemical-structure of ulvan. Carbohydr. Res., 274: 313–
318.
Ruberto G, Baratta M T, Biondi D M, Amico V (2001).
Antioxidant activity of extracts of the marine algal genus
Cystoseira in a micellar model system. J. Applied Phycol. ,
13: 403 407.
Savelieva I, Bajpai A, Camm J (2007). Apathophysiology,
new antithrombotic therapies, and evolution of
procedures and devices. Annals of Med., 39: 371-391.
Shayne C G (2005). Drug Discovery Handbook. New Jersey:
Wiley, (Chapter 1).
Sire O, Mangeney M, Montagne J, Nordmann R, Nordmann J
(1983). Carnitine palmitoyltransferase I. Inhibition by D-
Galactosamine and role of phospholipids. Eur. J.
Biochem.136 (2):371–375.
Tappel L, Zalkin H (1959) .Inhibition of lipid peroxidation
in mitochondria by vitamin E. Archives of Biochem. and
Biophysics. 80:333-336.
Taskin E, Ozturk M, Kurt O (2007). Antibacterial activities
of some marine algae from the Aegean Sea (Turkey). Afr.
Biotechnol., 6: 2746-2751.
Velichko MA, Shevchenko VP (1998). Biologically active
food additives. Voen. Med. Zh., 7: 24–27.
Yashizawa Y, Ametani A, Tsunehiro J, Numura K, Itoh M,
Fukui F, Kaminogawa S (1995). Macrophage stimulation
activity of the polysaccharide fraction from marine algae
(Porphyra yezoensis) . Biosci. Biotechnol. Biochem., 59:
1862–1866.
Zahra R, Mehrnaz M, Farzaneh V, Kohzad S (2007).
Antioxidant activity of extract from a brown alga,
Sargassum boveanum. Afri. J. Biotechnol. 6 (24):2740-
2745.
Zhang Q, Li N, Liu X, Zhao Z, Li Z, Xu Z. (2004). The structure
of a sulfated galactan from Porphyra haitanensis and its
in vivo antioxidant activity. Carbohydr. Res., 339 (1):
105– 111.

7. Int. J. Agric. Pol. Res. 378
with SAS software. (2nd ed.). John Wiley and Sons Co. pp:
1-340.
Lahaye M, Robic A (2007). Structure and functional
properties of ulvan, a polysaccharide from green
seaweeds. Biomacromolecules, 8:1765-1774.
Lau BHS (1998). Detoxifying radio-protective and
phagocyte-enhancing effects of garlic. International and
Nutrition Reviews, 9: 27-31.
Leeuwenburgh C, Heinecke J (2001) .Oxidative stress and
antioxidants in exercise. Current Med. Chem., 8: 829-838.
Liu D, Wong PTS, Dutka BJ (1973). Determination of
carbohydrates in lake sediment by a modified
phenolsulfuric acid method. Water Res., 7: 741-746.
McDermid KJ, Stuercke B (2003). Nutritional composition of
edible Hawaiian seaweeds. J. Appl. Phycol., 15: 513-524.
Msuya FE, Neori A (2002). Ulva reticulata and Gracilaria
crassa: macroalgae that can biofilter effluent from
tidalfishponds in Tanzania. West. Indian Ocean J. Mar. Sci.,
1: 117-126.
Mulbry W, Westhead K, Pizarro C, Sikora L (2005).
Recycling of manure nutrients: use of algal biomass from
dairy manure treatment as a slow release fertilizer.
Bioresource Technol., 96:451-458.
Nakanishi K (1999). An historical perspective of natural
products chemistry. In S. Ushio (Ed.), Comprehensive
Natural Products Chemistry. Elsevier Science B.V.,
Amsterdam, 1: 23–40.
Newman DJ, Cragg GM (2007). Natural products as sources
of new drugs over the last 25 years. J. Nat. Products,
70:461-477.
Norziah MH, Ching CY (2000). Nutitional composition of
edible seaweed Gracilaria changgi. Food Chemistry, 68:
69-75.
Pengzhan Y, Quanbin Z, Ning L, Zuhong X, Yanmei W, Zhi’en
L. (2003). Polysaccharides from Ulva pertusa
(Chlorophyta) and preliminary studies on their
antihyperlipidemia activity. J. Appl. Phycol., 15:21-27.
Percival E (1979).The polysaccharides of green, red and
brown seaweeds: their basic structure, biosynthesis and
function. Br. Phycol J., 14: 103-117.
Percival E, Wold JK (1963). The acid polysaccharide from
the green seaweed Ulva lactuca. Part II. The site of the
ester sulphate. J. Chem. Soc. 5459-5468.
Pourmorad F, Hosseinimehr J, Shahabimajd N
(2006).Antioxidant activity, phenol and flavonoids
contents of some selected Iranian medicinal plants. Afr. J.
Biochem., 5: 1142-1145.
Qi H, Zhang Q, Zhao T, Hu R, Zhang K, Li Z. (2006). In vitro
antioxidant activity of acetylated and benzoylated
derivatives of polysaccharide extracted from Ulva
pertusa (Chlorophyta). Bioorg. Med. Chem. Lett. 16:2441-
2445.
Qi H, Zhao T, Zhang Q, Li Z, Zhao Z , Xing R (2005).
Antioxidant activity of different molecular weight
sulfated polysaccharides from Ulva pertusa Kjellm
(Chlorophyta). Appl. Phycol., 17: 527-534.
Ray B, Lahaye M (1995). Cell-wall polysaccharides from the
marine green-alga Ulva-rigida (Ulvales, Chlorophyta) -
chemical-structure of ulvan. Carbohydr. Res., 274: 313–
318.
Ruberto G, Baratta M T, Biondi D M, Amico V (2001).
Antioxidant activity of extracts of the marine algal genus
Cystoseira in a micellar model system. J. Applied Phycol. ,
13: 403 407.
Savelieva I, Bajpai A, Camm J (2007). Apathophysiology,
new antithrombotic therapies, and evolution of
procedures and devices. Annals of Med., 39: 371-391.
Shayne C G (2005). Drug Discovery Handbook. New Jersey:
Wiley, (Chapter 1).
Sire O, Mangeney M, Montagne J, Nordmann R, Nordmann J
(1983). Carnitine palmitoyltransferase I. Inhibition by D-
Galactosamine and role of phospholipids. Eur. J.
Biochem.136 (2):371–375.
Tappel L, Zalkin H (1959) .Inhibition of lipid peroxidation
in mitochondria by vitamin E. Archives of Biochem. and
Biophysics. 80:333-336.
Taskin E, Ozturk M, Kurt O (2007). Antibacterial activities
of some marine algae from the Aegean Sea (Turkey). Afr.
Biotechnol., 6: 2746-2751.
Velichko MA, Shevchenko VP (1998). Biologically active
food additives. Voen. Med. Zh., 7: 24–27.
Yashizawa Y, Ametani A, Tsunehiro J, Numura K, Itoh M,
Fukui F, Kaminogawa S (1995). Macrophage stimulation
activity of the polysaccharide fraction from marine algae
(Porphyra yezoensis) . Biosci. Biotechnol. Biochem., 59:
1862–1866.
Zahra R, Mehrnaz M, Farzaneh V, Kohzad S (2007).
Antioxidant activity of extract from a brown alga,
Sargassum boveanum. Afri. J. Biotechnol. 6 (24):2740-
2745.
Zhang Q, Li N, Liu X, Zhao Z, Li Z, Xu Z. (2004). The structure
of a sulfated galactan from Porphyra haitanensis and its
in vivo antioxidant activity. Carbohydr. Res., 339 (1):
105– 111.