Evaluation of Antioxidant and Antimicrobial Activity in Relation

to Total Phenolic Content of Green Algae from Black Sea

RODICA SIRBU1, GABRIELA STANCIU2*, ANETA TOMESCU3, ANA MARIA IONESCU3, EMIN CADAR1
1
Ovidius University of Constanta, Faculty of Pharmacy, 124 Mamaia Blvd., 900527, Constanta, Romania
2
Ovidius University of Constanta, Department of Chemistry and Chemical Engineering, 124 Mamaia Blvd., 900527, Constanta,
Romania
3
Ovidius University of Constanta, Faculty of Medicine, 900527, 124 Mamaia Blvd., Constanta, Romania

Worldwide, marine algae are known to produce a wide variety of bioactive secondary metabolites and
several compounds have been derived from them for the prospective development of novel drugs by the
pharmaceutical industry. In this context, Ulva lactuca, Enteromorpha intestinales, and Cladophora vagabunda,
isolated from Black Sea coastal waters of Romania, were evaluated for their potential for bioactivity. Extracts
of the algae selected for the study were prepared using ethanol. The study presents original results concerning
the total phenolic content (TPC), determined by the spectrometric method (Folin-Ciocalteu method) and
the individual phenols from each species of seaweed studied were identified by HPLC-DAD techniques.
Total antioxidant activity (ACL) was studied by photochemiluminescence method, and antibacterial activity
was studied on two strains of bacteria, Escherichia coli ATCC 10536 - gram-negative and Staphylococcus
aureus ATCC 6538P - gram-positive. Results indicated that the highest total phenolic content was obtained
for Cladophora vagabunda (325.8 mg/100 g f.w.), followed by Ulva lactuca (285.5 mg/100 g f.w.) and the
lowest rate was registered by Enteromorpha intestinalis (268.8 mg/100 g f.w.). Through HPLC-DAD
techniques, 6 individual phenolic compounds were identified in all analyzed seaweeds. In addition, two
other phenolic compounds were identified in lower concentrations in Ulva lactuca, but only one phenolic
compound in Enteromorpha intestinalis. The results of this study show that there is a strong correlation
between antioxidant activity (ACL), antibacterial activity and total phenolic content (TPC). Antioxidant
activity shows the same order as total phenolic content (TPC): Cladophora vagabunda (64.15 nmol/g sample)
followed by Ulva lactuca (22.8 nmol/g sample) and Enteromorpha intestinalis (10.4 nmol/g sample). The
antibacterial activity recorded in all green seaweeds shows that the inhibition sites are higher on Escherichia
coli compared to Staphylococcus aureus.
Keywords: green seaweeds, antioxidant activity, antibacterial activity, total phenolic content, HPLC-DAD

The effects of light and the presence of oxygen and active various seas and oceans [16]. Marine algae have been
free radicals (O2, OH •, LOO •) on living systems has studied in the past years as a potential source of bioactive
become a frequently researched topic [1]. Oxidative compounds, due to the fact that they have the ability of
degradation of lipids from raw or processed food products producing a series of secondary metabolites with a variety
is not responsible only for the loss of nutritional value, but it of interesting biological effects, such as antimicrobial,
also contributes to carcinogenesis, atherosclerosis and the antiviral and antioxidant activity [17, 18]. There has been a
ageing process in humans [2]. Phenols are extremely particular interest in researching new antimicrobial
efficient antioxidants for polyunsaturated fatty acids; in substances originating from green marine algae from
fact, they can easily transfer a hydrogen atom to the peroxyl various seas and oceans [19-21]. Algae contain a high
lipid radical (LOO) thus forming the aryloxy radical (ArO), number of antibacterial compounds, such as fatty acids
which cannot act as chain carrier. This couples itself with [22], phenols [23], steroids [24, 25] and halogenated
another radical, thus stopping the chain process [3]. compounds [26, 27]. In the past years, algae extracts have
Amongst marine organisms, algae represent one of the been widely used in the prevention and treatment of viral
richest sources of antioxidants [4]. Phenolic compounds and bacterial disease in aquatic animals [28]. Algae
are present in all plants [5-7]. They can be found in extracts have shown a powerful antibacterial activity
significant quantities in algae, alongside other important against aquatic pathogenic bacteria [29]. From the green
active biologic compounds, such as polysaccharides, fatty algae, Ulva lactuca has been studied by numerous
acids, proteins, terpenes, and so on, which suggest researchers and it has become a model for morphogenetic
antitumor, antibacterial, antiviral and antioxidant activity studies [30]. Studies regarding green algae composition
[8, 9]. Marine algae are distributed globally in seas and were also performed by other researchers who have taken
oceans and can be found attached to rocks or other hard into account the seasonal changes in compositions for Ulva
substrates from coastal regions, with an 0.01% availability lactuca [31]. Other studies have determined the chemical
of photosynthetic light [10]. These can be found in every composition of ulvan [32, 33] and have isolated extracts
climate area, from warm tropical waters to polar, freezing with antioxidant and antibacterial activity [34-36]. The
regions [11]. Green algae can be found worldwide, growing Black Sea, like other seas and oceans, offers a rich algae
in intertidal zones, attached to a hard substrate or free [12]. mass which can be harvested, thus representing an
These are considered opportunistic algae and they important resource for the medical and pharmaceutical
proliferate in eutrophic coastal waters [13]. There are fields [37]. Until now, studies regarding the characterization
numerous studies regarding green algae [14, 15]. Thus, of marine algae, which have been used in various purposes,
phenols and active biologic compounds have been found have been performed [38]. Regarding Black Sea algae,
in several green algae species which can be observed in marine habitats in which algae develop have been studied,

email: gstanciu66@[Link], Phone: 0040726182412 All authors are considered main authors with equal contributions
[Link].(Bucharest)♦70 ♦ No. 4 ♦ 2019 [Link] 1197
especially in the Romanian shore area [39]. However, the the same conditions as mentioned above. The combined
antioxidant and antimicrobial activity of the seaweed filtrate was evaporated in a rotary evaporator (BUCHI)
remains unexplored. The main objective of this paper is to below 40 0C. The extract obtained after evaporation of the
evaluate the total phenol content and its correlation with organic solvent was weighed for calculation of extraction
antioxidant and antimicrobial activity of Clorophyta green yield and was used as a natural antioxidant.
algae. The following species have been selected: Ulva
Lactuca (Linnaeus, 1753) [40] and Enteromorpha Determination of total phenolic content (TPC)
intestinales (Linnaeus-Nees, 1820) from the Ulvophyceae Total phenolic contents in extracts were determined
class, Ulvales order, Ulvaceae family and Cladophora according to the Folin - Ciocalteu method [42, 43] using a
vagabunda (Linnaeus-van den Hoek, 1963) [41], from the UV spectrophotometer ( Cintra Xe ) according to a
Chlorophyceae class, Cladophorales order, colorimetric reduction of a phosphowolframate -
Cladophoraceae family. Phenolic content in alcoholic phosphomolibdate complex to blue products in sodium
extracts from algae harvested from six different regions of carbonate media. The absorbance relative to a gallic acid
the Romanian shore was determined, and the antioxidant standard curve was measured and results are expressed
(ACL) and antimicrobial activity of the algae was as gallic acid equivalents (mgGAE/L). All samples were
evaluated. The total phenolic content was determined performed in triplicate and the mean value was reported.
through the Folin-Ciocalteu methods. Phenolic compounds
from green algae extracts were determined through HPLC- Calibration curve
DAD techniques. Antioxidant capacity (ACL) was The calibration curve (fig. 1) was graphically represented
evaluated through the chemiluminescence method, using by mixing 1 mL aliquots of 50, 100, 150, 200, 250, 300, 350,
trolox as the standard substance. Green algae antibacterial 400 and 450 mg/L gallic acid solutions with 5.0 mL of Folin-
activity was evaluated on two strains of bacteria: gram Ciocalteu reagent (1:10) and 4.0 mL of sodium carbonate
negative Escherichia coli and gram positive solution (75g/L). Absorbance was measured after 30 min
Staphylococcus aureus. at 765 nm. In order to determine total phenolic content
from ethanol extracts, 1 mL of sample was mixed with
Experimental part the same reagents as the ones used for the calibration
Plant materials curve.
The samples of green seaweed were collected during
July-Octomber 2017, when algae reach maturity in different
harvesting sites. Samples were harvested from 6 areas,
and named as: S1 through S6: S1- Mamaia North, S2 -
Constanta Casino, S3 -Constanta harbor, S4 -Eforie South,
S5 -Costinesti, S6- Mangalia. Each collected sample was
thoroughly washed with seawater to remove all the
extraneous matter such as epiphytes, sand particles,
pebbles and shells. In the laboratory the sample was
cleaned with fresh water and distilled water, then dried
and powdered using mixture grinder. The powdered
seaweeds were passed through a 0.5 mm sieve in order to Fig.1. Calibration curve of gallic acid standard
obtain a uniform powder which was stored in the freezer The amount of total phenol contents was calculated as
until extraction. Gallic acid equivalents in mg/100 g of dried seaweed (mg
GAE/100 g f.w.).
Chemicals
All used reagents were of analytical reagent grade. Gallic Identification of phenolic acids by HPLC
acid was purchased from Fluka and Folin - Ciocalteu Identification of phenolic acid was performed through
reagent from Merck. Gallic acid (standard phenol HPLC-DAD method. The extract was passed through a 0.45
compound) 1×10-2 mol×L-1 solution was prepared by µm filter (Millipore) before being injected into the HPLC.
dissolving 376 mg of gallic acid in 100 mL of ethanol. Folin Reverse phase HPLC was performed with an Agilent 1100
- Ciocalteu reagent was diluted with distilled water 1:10 series HPLC (Agilent Technologies), equipped with a DAD
(v:v). The gram positive and gram negative bacteria strains detector (Agilent G13158). The column used was a ZORBAX
were grown on culture media. Eclipse XDB C8 analytical column (150 x 4.6 mm) with a
5µm packing material. The elution was performed with an
Apparatus isocratic mixture of methanol and 10 mM ammonium
Spectrometric measurements were carried out using a acetate buffer, pH 5.4 (12:88 v/v) at a flow of 1 mL/min.
UV-Vis Cintra 10e scanning spectrophotometer. The Detection was done using a diode array detector (DAD)
identification and quantitative determination of phenolic with reference wavelength of 280 nm. Retention times
compounds were performed using HPLC system (Agilent and peak areas were monitored and computed
1200) with quaternary pump, DAD, auto sampler. The automatically by Chem32 integrator (Agilent). The
antioxidant capacity of lipid soluble substances standard reference substances chosen were gallic acid,
determination was performed with a Photochem protocatechuic acid, gentisic acid, p-hydroxybenzoic acid,
instrument with ACL kit (Analytic Jena ∆G). vanillic acid, caffeic acid, salicylic acid, benzoic acid, ferulic
acid, elagic acid and p-coumaric acid, for which a 0.05
Preparation of algal extract mg/mL concentration was established. Standards were
Ethanol was used as an extraction solvent. For the provided by Sigma Aldrich. Individual phenolic acids were
preparation of the ethanol extract, 5 g of powdered identified by comparing the retention time of sample
seaweed was extracted overnight with 50 mL of 70 % chromatographic peaks with those of authentic standards
ethanol (v:v) at room temperature and centrifuged at 2800 using the same HPLC operating conditions. Table 1
rpm for 10 min. The supernatant was collected in a separate presents the determined retention time of standards.
bottle and the residue was re-extracted three times under
1198 [Link] [Link].(Bucharest)♦70♦ No. 4 ♦ 2019
 Table 1 The calibration curve was designed through the
THE RETENTION TIME OF STANDARDS measurement of a series of standard solutions containing
0.5, 1.0, 2.0, 3.0 and 4.0 nmol trolox and is presented in Fig.
2. Photochemical reaction was generated by exposure to
external radiation from a Hg lamp which yields maximum
energy at λ= 351 nm. Total antioxidant capacity of the
sample was measured by comparison to the trolox
standard, in standard equivalent units [45].

Antibacterial activity
In order to test antimicrobial activity of the green
seaweed, we used several culture media. We worked on
two types of bacteria: gram-positive Staphylococcus
aureus ATCC 6538P and gram-negative Escherichia coli
ATCC 10536. The analysis method used was the nutrient
agar well diffusion method. The methodology involves the
preparation of nonspecific nutrient agar plates, filled with
a known concentration of bacterial medium and measuring
the inhibition halos formed by the ethanol extract solutions
from green seaweed. Algae species were washed, dried
Statistical analysis and brought to a corresponding grinding degree. Dried algae
All experiments were conducted in triplicate and ANOVA were subjected to ethanol extraction (5 g powder per 100
(using SPSS 11.5 statistics) software was used to compare mL ethanol) through continuous stirring at 37 0C, for 24 h.
the mean values of each treatment. Significant differences The samples were subjected to centrifugation for 30
between the means values of parameters were determined minutes and the supernatant was evaporated under
by using the Duncan test (P < 0.05). vacuum. The extracts were re-dissolved in ethanol.

Determination of total antioxidant activitiy (ACL) Preparation of nutrient agar plates

The ACL assay was performed using Photochem Culture media was prepared for each strain,
instrument (Analytic Jena) against the superoxide anion Staphylococcus aureus and Escherichia coli (fig.3). Petri
radicals from luminol as photosensitizer, upon the UV light dished in which 1 mL reference solution (Inocculum) and
[44]. In order to determine total antioxidant capacity (ACL) 80 mL of nonspecific nutrient agar were added, sterilized
of biological samples, the well dried algae were triturated and homogenized (fig.3). The plates were left to cool at
to a fine powder, after which a cold extract was obtained room temperature until the agar became solid. Two plates
from each alga species, using 20 mg of dried vegetal with positive witness samples were also prepared in order
product dissolved in 1 mL ethylic alcohol p.a. [45]. to compare results.
Extraction was performed for 24 h at room temperature in
yellow glass sealed bottles. Mixtures were stirred regularly
and extraction separation was performed through
decantation, without filtration. Antioxidant capacity was
determined using the standardized ACL method
(antioxidant capacity of lipid soluble substances, Analytik
Jena 10) in comparison to the standard substance trolox Fig.3 Culture media
(brand name Hoffman-La Roche-6-hydroxy-2, 5, 7, 8- for Staphylococcus
tetramethylchromen-2-carboxylic). For the calibration aureus and
curve, an Analytic Jena standard reagent kitts was used: Escherichia coli
R1 (dilution solvent), R2 (buffer reagent), R3
(photosensitive reagent), R4 (reagent dimension).

For each of the strain, a standardized reference solution

with a known concentration (3 x 10-8 g/mL) was prepared.
After solidification of the agar from the Petri dishes the
wells were made, using a sterilized matrix with a known
diameter, corresponding to each solution volume. Thus,
for a 0.25 mL volume, was a matrix with a diameter of 10
mm was used, whereas, for a 0.5 mL volume, we used a
matrix with a diameter of 15 mm, and for a 1 mL volume,
the a diameter was of 17 mm. Interpretation of the results
of this antimicrobial analysis was performed through the
Fig. 2. Calibration curve for trolox in the calculation of trolox comparison of the obtained samples with the positive
equivalents for ACL measurements (generated by PCLsoft) witness plate for each studies pathogen culture. The
diffusion radius of the green algae extract solution in the
agar contaminated with the study strain was measured.
[Link].(Bucharest)♦70 ♦ No. 4 ♦ 2019 [Link] 1199
The halo formed around the wells represents growth HPLC analysis of phenol compounds
inhibition and the halt of pathogen spread; the radii differ Table 3 presents the individual phenolic concentrations
according to the type of alga, as well as the concentration determined by HPLC -DAD expressed in mg/100 g f. w. By
and the solution volume from the algae extracts. correlating the total phenolic content (TPC) and individual
phenols the majority phenolic content can also be
Statistical analysis expressed as percentage (table 4). It can be observed that
A statistical approach was designed and the the total phenols identified and quantified were in different
experimental data were evaluated using one-way analysis percentages for the ethanol extracts from the three algae.
of variance (ANOVA), with p < 0.05 as threshold for The highest percentage of individual phenols was identified
statistical significance. The statistical results confirm the in Ulva lactuca (79.96%), followed by Cladophora
hypothesis that the differences between the results are vagabunda (74.6%), while the lowest percentage was
either not significant (p > 0.05). The average of multiple obtained for Enteromorpha intestinalis (67.93%). Table 3
measurements (triplicates or more) was listed in the tables shows that all three Clorophytele green algae contain
together with the standard deviations. Statistical analysis gentisic acid; the highest quantity was identified in
was performed using Excel software package. Cladophora vagabunda (155.3 mg/100 g f.w.), followed by
Ulva lactuca (130.5 mg/100 g f.w.) and Enteromorpha
Results and discussions intestinalis (98.6 mg/100 g f.w.). If we correlate the total
Total phenol content (TPC) phenol content (TPC) identified and quantified in the
Three different green seaweed from the Clorophyta class analyzed seaweed with the gentisic acid content found
were studied: Ulva lactuca, Enteromorpha intestinales and (see Table 4), it can be observed that this acid has the
Cladophora vagabunda. The results of total phenolic highest percentage of total existing phenols in Ulva lactuca
content (TPC) determined for ethanol extracts of these (46.0%), followed by 43.52% in Cladophora vagabunda and
green algae are presented in table 2. 36.68% in Enteromorpha intestinalis.
The total phenolic content ranged from 268.8 to 325.8 Protocatechuic acid is another phenol compound found
mg/100 g dried seaweed in ethanol. The lowest, but still in considerable amounts in green seaweed ethanolic
rich, total phenolic content was obtained for Enteromorpha extracts. This acid was found in Cladophora vagabunda
intestinalis (268.8 mg/100 g f.w.). The highest total phenolic (75.9 mg/100 g f.w.), in Ulva lactuca (72.8 mg/100 g f.w.)
content was obtained for Cladophora vagabunda (325.8 and the least in Enteromorpha intestinalis (55.6 mg/100 g
mg/100 g f.w.) followed by Ulva lactuca (285.5 mg/100 g f.w.).
f.w.). Table 4 outlines the following percentage order in which
There are studies in literature which outline the fact that protocatechuic acid was found: 25% in Ulva lactuca,
the type of solvent influences selectivity of phenol 21.27% in Cladophora vagabunda and 20.68% in
compounds [42]. The study performed by Wang et al. [23], Enteromorpha intestinalis from the total content of
shows that water as solvent is inferior to polar organic polyphenols identified in each alga.
solvents in the extraction of polyphenolic compounds,
which confirms our use of ethanol as a solvent.

Table 2
TOTAL PHENOL CONTENT (TPC)
OF ETHANOLIC EXTRACTS OF
GREEN SEAWEEDS FROM THE
ROMANIAN BLACK SEA COAST

Table 3
INDIVIDUAL
PHENOLIC
CONCENTRATIONS
DETERMINED BY
HPLC -DAD IN GREEN
SEAWEED FROM THE
ROMANIAN BLACK
SEA COAST

1200 [Link] [Link].(Bucharest)♦70♦ No. 4 ♦ 2019

 Table 4
THE PERCENTAGES (%) OF TOTAL PHENOLS AND PERCENTAGES (%) OF MAJOR PHENOL COMPOUND IDENTIFIED IN TESTED ALCOHOLIC
BERRIES EXTRACTS DETERMINED BY HPLC-DAD REPORTED TO TPC VALUES

Another acid found in significant amounts is represented The highest ACL values were registered by Cladophora
by caffeic acid: in Enteromorpha intestinalis (15.7 mg/100 vagabunda (64.15 nmol/g dry alga), followed by Ulva
g f.w. with a percentage of 5.84%), in Ulva lactuca (13.0 lactuca (22.8nmol/g dr y alga) and Enteromorpha
mg/100 g f.w. with a percentage of 4.55%) and in intestinalis (10.4 nmol/g dr y alga). This order is in
Cladophora vagabunda (11.7 mg/100 g f.w with a accordance with the total polyphenol content (TPC) in the
percentage of 3.28%). It has been noted that in Cladophora analyzed algae: Cladophora vagabunda (325.8 mg/100g
vagabunda, the vanillic acid (12.8 mg/100 g f.w) was found f.w.), followed by Ulva lactuca (285.5 mg/100 g f.w.) and
in comparable amounts with caffeic acid (11.7 mg/100 g Enteromorpha intestinalis (268.8 mg/100 g f.w).
f.w). The remainder phenolic acids which were identified The results of this study confirm the fact that there is a
and quantified through HPLC-DAD, were found in smaller strong correlation between total antioxidant activity (ACL)
concentrations. Due to different extraction methods and and total polyphenol content (TCL).
different species studied, as well as the measurements It is believed that the antioxidant properties of
used and the lack of information regarding phenolic polyphenols are the result of their ability to act as reducing
composition of Black Sea algae in literature on the matter, agents, hydrogen donors and free radicals and of caffenol’s
a direct comparison between our results and studies function as metal chelator, which impede the catalytic
performed in other seas is not feasible. functioning of metal in the radical initiation process [16,
46].
Antioxidant activity (ACL)
It is very important to examine the correlation between Antibacterial activity
the total phenol content and the total antioxidant activity The antibacterial activity of the three green algae is the
(ACL) in the studied algae. The extraction method used to result of their high phenolic compound concentrations.
obtain fluid extracts was cold macerated in 70% Figure 5 and figure 6 present the inhibition halos formed as
concentration ethylic alcohol, for 24 h in the dark. A good a result of the own antimicrobial action exercised by each
total antioxidant capacity was generated, as it can be seen alga extract ( Cladophora vagabunda, Ulva lactuca,
from the curves obtained using the Photochem instrument Enteromorpha intestinales ), on bacterial strains of
with ACL kitt (fig.4). Table 5 presents the results for ACL Escherichia coli and Staphylococcus aureus.
expressed as Trolox units equivalents (nmol/g of dry It can be observed that the analyzed alga extracts
sample) for the three algae. exhibit different radii for bacterial growth inhibition for each

Fig. 4 Antioxidant
activity (nmoles trolox /
mg fresh weight) for
undiluted sample and
diluted samples 1:25
measured using
PHOTOCHEM
equipment

Table 5
RESULTS OBTAINED
FOR THE ACT
DETERMINATION ON
GREEN ALGAE
EXTRACTS

[Link].(Bucharest)♦70 ♦ No. 4 ♦ 2019 [Link] 1201

 Fig.5 The antimicrobial
effect of alcoholic alga
extracts on Escherichia coli

Fig.6 The antimicrobial effect of

alcoholic alga extracts on
Staphylococcus aureus

Fig.7. Antibacterial Activity of ethanolic extracts from green seaweeds in the Romanian Black Sea coast on strains
of bacteria (Escherichia coli and Staphylococcus aureus)
of the three algae. This antimicrobial effect, evidenced [52] have shown that there is an evident seasonal variation
through the size of the inhibition areas is an important in antibacterial activity of green algae harvested from
property of seaweed, of great use in the pharmaceutical China’s territorial waters [52]. Also, Chinese researchers
field. Figure 7 shows the graphical representation of have observed that Ulva lactuca has the highest
antimicrobial activity of seaweed extracts on the analyzed antibacterial activity from the beginning of winter
pathogen germs. Inhibition radii obtained for the studied (December) until mid-spring (April) [52].
algae extracts are similar in value, with a different
antimicrobial effect on the bacterial strains that can be Conclusions
observed. In our study the highest activities of ethanol This is the detailed study of in vitro antioxidant and
extracts (10.5 mm, 12.2 mm and 16.3 mm) were recorded antibacterial activity of seaweeds from the Romanian
for Escherichia coli and the lowest (4.6 mm, 5.9 mm and Black Sea coast in which phenolic compounds were
7.8 mm) for Staphylococcus aureus at 100 µg/mL. identified and quantified from a range of green seaweeds.
Although literature on the matter holds few data regarding The results of this screening experiment demonstrated that
antibacterial activity of green alga in the Black Sea, our extracts of the different seaweed species contain different
results are in accordance with the antibacterial activity of levels of total and individual phenolics and possess diverse
other green seaweed species ( Ulva reticulate and antioxidant and antibacterial properties.
Cladophora socialis) in ethanolic extracts that can be found Three green algae species from the Clorophyta class
in the Red Sea [47]. In literature, the maximum activity were analyzed: Ulva lactuca (L.), Enteromorpha intestinales
was recorded in ethanol and ethyl acetate extracts from (L-Nees) and Cladophora vagabunda (L-Hoek). The total
seaweeds and both of these extracts exhibited reasonable phenolic content (TPC) as determined by spectro-
activity against all the tested pathogens [48, 49], which photometry (the Folin-Ciocalteu method) was found in all
confirms our choice of solvent. Studies performed on studied algae, in varying concentrations. The highest total
different types of seaweed originating from the territorial phenolic content was obtained for Cladophora vagabunda
South African waters have been reported, in which the (325.8 mg/100g f.w.), followed by Ulva lactuca (285.5 mg/
antibacterial activity manifests itself all throughout the year 100g f.w.) and Enteromorpha intestinalis (268.8 mg/100g
at different intensities [50]. f.w.).
Other researchers claim that the activity of algae Using HPLC-DAD techniques 6 individual phenol
originating from India’s territorial waters can be due to the compounds were found, in varying quantities, in all the
variation in cellular chemical and biological composition analyzed algae. High contents were obtained for: gentisic
according to season, habitat, and life stage of the respective acid (98.6-155.3 mg/100 g f.w.), protocatechuic acid 55.6-
alga [51]. Recent studies performed by Chinese researchers 75.9 mg/100 g f.w.), caffeic acid (11.7-15.7 mg/100 g f.w.),
1202 [Link] [Link].(Bucharest)♦70♦ No. 4 ♦ 2019
gallic acid (5.3-8.5 mg/100 g f.w.), and in lower contents [Link], K.J., ELMANAMA, A.A., CHIHADEH, W.N, J. Mar. Sci., 28,
for p-vanillic acid (1.6-12.8 mg/100 g f.w.) and p- no.1, 2013, p. 81.
hydroxybenzoic acid (1.5-2.6 mg/100 g f.w.). Moreover, [Link], E., OZTURK, M., TASKIN, E., KURT, O., African J. Biotech.,
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[Link], J.K., RATH, S.K., JENA, K., RATHOD, V.K., THATOI, H.N.,
(TCL). Antioxidant activity, as determined through
Turkish J. Biol, 32, 2008, p. 119.
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phenolic content: Cladophora vagabunda (64.15 nmol/g Tech Res., 6, no.1 2014, p. 1.
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of yielding pharmaceutically useful algae, which can be [Link], T., CHARRIER, B., MINEUR, F., BOTHWELL, F.H., DE
harnessed for the development of drugs for use in CLERCK, O., COATES, J. C., Frontiers in Plant Science, 6, 2015, p. 72.
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for the financial support of the Romanian Ministry of Research - [Link], T., ENGELEN, A. H., DIEKMANN, O.E., KUEGLER, S.,
UEFISCDI, for Complex Project no. 85 PCCDID 2018, Acronym WICHARD, T., Chem. Phys. Lipids, 183, 2014, p. 100.
INOBIOMAR, Project code PN-III-P1-1.2-PCCDI-2017-0701. [Link]ÁNDEZ-GARIBAY, E., ZERTUCHE-GONZÁLEZ, A., PACHECO-
RUIZ, I., J. Appl. Phycol, 23, no.3, 2011, p. 537.
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