analysis of chemical compounds by using gas ... - …

ANALYSIS OF CHEMICAL COMPOUNDS BY USING GAS CHROMATOGRAPHY

AND MASS SPECTRUM ANALYSIS, IN VITRO ANTIOXIDANT AND

ANTIBACTERIAL ACTIVITY OF METHANOLIC EXTRACTS OF SEAWEED

ULVA FLEXUOSA WULFEN (GREEN ALGAE)

Manjula Duraisamy1 & Raja Selvaraju*2

Research Scholar, Department of Zoology, Kongunadu Arts and Science College, Coimbatore, Tamilnadu, India1

Assistant Professor, Department of Zoology, Kongunadu Arts and Science College, Coimbatore, Tamilnadu, India2

[email protected]; [email protected]

*Corresponding author

ABSTRACT: In the present study Ulva flexuosa was collected along the shore of

Mandapam and was identified and authenticated. To analyse the methanol extract of marine

green macro algae species Ulva flexuosa using Gas chromatography-Mass spectrometry

(GC-MS), in vitro antioxidant and antibacterial activity. Gas chromatography and mass

spectrometry analysis of whole plant extract injected with instrument GC-MS-QP 2010

[SHIMADZU], In vitro antioxidant activities were determined by the DPPH (1, 1-

diphenylpicryl- 1-picryl-hydrazyl) radical scavenging and Metal chelating assays. The total

phenol content in methanol extracts was determination as 18.5±0.5 mg/g and flavonoids

contents were 46±3 mg/g. The methanol extracts of Ulva flexuosa was identified 22 bioactive

compounds which were major compound such Hexadecanoic acid, methyl ester (29.10%),

13-Docosenamide, (Z) (10.27%), Eicosane (10.24%) Cholest-5-en-3-ol, 24-propylidene-,

(3á)- (9.00%), 9-Octadecenoic acid (Z)-, methyl ester (CAS) (8.07%). These bioactive

compounds could be a valuable for agricultural purposes, animal feed preparation and drug

making in pharmaceutical industries.

Key words: Ulva flexuosa, GC-MS analysis, In vitro antioxidant activity, antibacterial activity, prebiotic

INTRODUCTION

Marine seaweeds have been harvested for several years in the Far East and Asia

Pacific countries, where they are consumed as food. In particularly the genus Ulva is widely

distributed around the world and tolerant to some environmental challenges, enabling the

amount of biomass generated being in order of tons. Around 16 million tons of seaweeds

(fresh weight basis) and other marine plants are annually produced or collected with an

estimated value of 5575 million Euros worldwide (FAO. 2016).

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http://aegaeum.com/ Page No: 1437

Ulva flexuosa is belonging to the family Ulvaceae and it is used as a bioindicator for

metal contamination, including lead and iron (Ho, Y.B. 1987; Tabudravu et al., 2002). U.

flexuosa is a species with hollow one layered thalloid green alga which grows rocks and

shores in the shallow water. The cell wall of Ulva contains polysaccharides called ulvan,

which comprises rhamnose, sulfate, xylose, iduronic acid, galactose, and glucose (Robic et

al., 2009a). Ulvan can elicit responses and induced defence mechanism in cultivated

organisms (Borsato et al., 2010). Ulva flexuosa was also found to be an economically

efficient species for bio-absorption in industrial settings. It could be used as an eco-friendly

alternative for wastewater treatment in dye manufacturing, tannery, textile, and cosmetic

industries (Sivasamy et al., 2012). Recently, much effort is paid to explore the contribution of

Ulva in development of novel drugs, pharmaceutical and agricultural applications (Costa et

al., 2010; Wijesekara et al., 2011). Macroalgae, especially Ulva species being an efficient

nutrient pump have high bioremediation efficiency and are suggested to reduce

eutrophication in aquaculture (Neori et al., 2003; Zhou et al., 2006). The aim of this study is

to analyse the methanol extract of marine green macro alga Ulva flexuosa using Gas

chromatography-Mass spectrometry (GC-MS), in vitro antioxidant and antibacterial activity.

MATERIALS AND METHODS

SAMPLE COLLECTION

The seaweed, Ulva flexuosa (Figure 1) was collected from the intertidal shallow zone at

depth of 0-1 m at Mandapam, Ramanathapuram, Tamil Nadu. The alga was obtained from the

Mandapam coast, Gulf of Mannar region, Rameswaram (Latitude: 9°16’32.56” N and

Longitude: 79°7’25.03” E) along the southern regions of Tamil Nadu. The sample of seaweed

was identified self and binomially by Botanical Survey of India (Southern part Coimbatore,

Tamilnadu, India) and voucher specimen (BSI/SRC/5/23/2018/Tech.1383) was deposited at

the Herbarium Department of zoology, Kongunadu Arts and Science College (Autonomous),

Coimbatore, Tamilnadu, India.

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Figure 1. Seaweed Ulva flexuosa (Green Algae)

EXTRACTION OF THE MATERIAL

50g of Ulva flexuosa fine powder was packed with Whatman No.1 filter paper and

placed in soxhlet apparatus along with solvent methanol. The residues were collected and

dried at room temperature, 30˚C after which yield was weighed and then performed to

activity.

DETERMINATION OF TOTAL POLYPHENOL AND FLAVONOID

The total phenol content was determined by the method of Siddhuraju and Becker,

(2003). Flavonoid contents were determined according to the method (Zhishen et al., 1999).

The analysis was performed in triplicate and the results were expressed as the gallic acid

equivalents (GAE).

IN VITRO ANTIOXIDANT ACTIVITY

DPPH· RADICAL SCAVENGING ACTIVITY

The 2, 2-diphenylpicryl- 1-picryl-hydrazyl (DPPH·) radical scavenging activity of

entire plant extract was measured according to the method (Blios, 1958). IC50 values of the

extract i.e., concentration of extract necessary to decrease the initial concentration of DPPH

by 50% was calculated.

METAL CHELATING ACTIVITY

The metal chelating effect on ferrous ion was determined according to the method

of Dinis et al., (1994). The metal chelating activity of the extracts was evaluated using

(Ethylene Diamine Tetra acetic acid) as standard. The results were expressed as mg EDTA

equivalent/g extracts.

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TOTAL ANTIOXIDANT ACTIVITY BY THE ABTS·+ ASSAY

Total antioxidant activity was determined according to the method of Re et al., (1999). The

unit of total antioxidant activity (TAA) is defined as the concentration of Trolox having

equivalent antioxidant activity expressed as μmol/g sample extracts on dry matter.

GAS CHROMATOGRAPHY AND MASS SPECTRUM ANALYSIS

5 ml of methanol extract was evaporated to dryness and reconstituted into 2 ml

methanol. The extract was then subjected to GC-MS analysis. Chromatographic separation

was carried out with instrument GC-MS-QP 2010 [SHIMADZU] instrument with Db 30.0

column (0.25µm diameter × 0.25um thickness). The oven temperature was programmed from

70 °C (isothermal for 5 min), with an increase of 10°C/min, to 200°C, then 5°C/min to

280°C, ending with a 35 min isothermal at 280°C. Mass spectra was taken at 70 eV; a scan

interval of 0.5 s and Scan range from 40–1000 m/z. Helium was used as carrier gas at

99.999% pressure with flow 1.0 ml/min and electronic pressure control on. Sample was

dissolved in methanol and injected automatically.

ANALYTICAL CONDITION

Injection temperature at 2400C, interface temperature at 2400C and ion source

temperature at 700C were determined. Injection was performed in split less mode. The mass

spectra of compounds in samples were obtained by electron ionization (EI) at 70 eV and the

detector operator in scan mode from 40 to 1000 m/z atomic mass units. Identification based

on the Molecular weight, Molecular formula, Retention time and peak area %.

IDENTIFICATION OF COMPOUNDS

Identification was based on the active principles with their Retention time (RT),

Molecular formula (MF), Molecular weight (MW) and concentration (peak area %). It is

done in order to determine whether this plant species contains any individual compound or

group of compounds which may substantiate its current commercial and traditional use as

herbal medicine, in addition to determine the most appropriate methods of extracting these

compounds. These results will consequently be discussed in the putative biological and

therapeutic relevance.

IN VITRO ANTIBACTERIAL ACTIVITY

Disc diffusion method (Rios et al., 1988; Collins et al., 1995) for assessing the

antimicrobial activity was measured around disc. 10µl of plant extract (40mg/0.1ml) was

soaked by sterile filter paper discs (6mm in diameter). The sterile filter paper discs were

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impregnated with plant extract placed on the surface of the medium the standard antibiotic as

tetracycline and incubated at 37˚C for 24 h.

STATISTICAL ANALYSIS

Data were expressed as mean, standard error and standard deviation of three replicate

determinations using Microsoft Excel windows 16.

RESULTS AND DISCUSSION

PHENOL FLAVONOIDS COMPOUND

Phenol compound is ubiquitous bioactive compounds and a diverse group of

secondary metabolites universally present in higher plants (Liu et al., 2009). The phenolic

compounds may contribute directly to anti oxidative action. In the present study the macro

algae U. flexuosa methanolic extracts showed that the highest phenolic content 18.5±0.5

mg/g are GAE g-1equalant (Table 1). The methanol is good solvent for Ulva flexuosa as large

amount of phenolics compounds. The Phenolic compounds are known as powerful chain

breaking antioxidants (Shahidi et al., 1992). The results showed that Ulva flexuosa contain

methanol extract has showed highest 46±3 mg/g (Table- 1). Some flavonoids were reported

to exhibit potential for anti–human immune deficiency virus functions (Yao et al., 2004).

Table 1. Polyphenol content of methonolic extracts of green macro algae U. flexuosa.

S. No Sample extracts Total Phenol

(mg GAE g-1) Total Flavonoids

(mg/g)

1 Methanol 18.5±0.5 46±3

All the values are expressed as mean ± SD (n=3).

IN VITRO ANTI-OXIDANT ACTIVITY DPPH

(1, 1-DIPHENYL-2-PICRYLHYDRAZYL) RADICAL SCAVENGING ACTIVITY

In the present study the green alga Ulva flexuosa results on DPPH radical scavenging

activity of methanol extracts were presented in Figure 2. Therefore, lower IC50 indicated a

higher antioxidant activity. Methanolic extract of (36.72 μg/mL) plant extracts showed higher

levels of free radical scavenging activity compare to the standard drug (25.9 μg/mL)

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Butylated Hydroxy Toluene (BHT). Likewise, previous reports have been demonstrated that

green algae have high antioxidant potential and scavenging activity. Several studies have

been demonstrated that the correlation between the phenolic content and antioxidant activity

of certain seaweeds (Siriwardhana et al., 2003). Luo et al. (2010) have demonstrated that

phenolic compounds were one of the most effective antioxidants in marine algae (Zakaria et

al., 2012). Flavonoids are more stable, less-reactive when they oxidized by radicals. A

positive correlation has been documented between anti-oxidation capabilities and total

polyphenol contents, but not with the contents of flavonoids (Liu et al., 2010).

Figure: 2. DPPH radical scavenging activity of methanolic extracts Ulva flexuosa.

METAL CHELATING ASSAY

The results of experiment conducted to assess the metal chelating activity of Ulva

flexuosa were presented in Table 2. The methanolic extract was displayed an apparent

antioxidant activity as they were able to chelate (30.62 mg/g), then the standard drug

EDTA.

ABTS•+ RADICAL SCAVENGING ASSAY

The ability of the test sample to scavenging ABTS

+ radical cations was equivalent of

Trolox solution, having a total antioxidant ability equivalent to 1g dry weight of the extract

under the experimental investigation. The highest ABTS radical scavenging rate was found in

the methanol extract as 2863.19 µmol/g (Table 2). Table. 2: Metal chelating activity and

TAA by ABTS+ assay of methanol extracts of U. flexuosa

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All the values are expressed as mean±SD (n=3).

GC-MS ANALYSIS

The GC- MS studies in the methanolic extract of green alga Ulva flexuosa showed the

presence of rich variety of phytochemical compounds, the active principles with their

retention time (RT), Molecular formula, molecular weight (MW), concentration (peak area

%) are presented in (Figure 3 and Tables 3 & 4). In the present study GC-MS analysis totally

twenty two compound were identified and revealed that existence of the major compound

such as Hexadecanoic acid, methyl ester (29.10%) (Figure 4) 13-Docosenamide, (Z)

(10.27%) (Figure 5), Eicosane (10.24%) (Figure 6), Cholest-5-en-3-ol, 24-propylidene-, (3á)-

(9.00%) (Figure 7), 9-Octadecenoic acid (Z)-, methyl ester (CAS) (8.07%) (Figure 8), Methyl

tetradecanoate (3.30%), 3,6-bis(t-Butyl) fluorenone (3.13%), Hexadecanoic acid (CAS)

(2.23%), N-BZ-2amino cinnamate (1.67%), Lucenin 2 (1.43%), 5,7,9(11)-Androstatriene, 3-

hydroxy-17-oxo- (1.35%), 2-Pentadecanone, 6,10,14-trimethyl- (CAS) (1.33%), Tetraneurin -

A – diol (1.23%), Tetradecane (CAS) (1.15), Hexa-t-butyl selenatrisiletane (0.99%),

Dotriacontane (CAS) (0.92%), 2,3-Dihydroxypropyl elaidate (0.72%), Hexadecanoic acid,

2,3-dihydroxypropyl ester (CAS) (0.69%), 9,12,15-Octadecatrienoic acid,2,3-

bis[(trimethylsilyl) oxy] propyl ester, (Z, Z, Z) (0.65%), 2,2,3,3,4,4 hexadeuterooctadecanal

(0.62%), 7-Methyl-Z-tetradecen-1-ol acetate (0.54%) and Z-(13,14-Epoxy) tetradec-11-en-1-

ol acetate (0.54%) (Figure 9).

The identified active phytocompounds are proven to possess pharmacologic activities

which may contribute to the healing potential of the seaweed. In previously reported that Gas

Chromatography-Mass Spectrum analysis in different macro algae Scinaia bengalica (Lalitha

and Palani, 2017), Gracilaria corticata (Jenifer et al., 2018), Gracilaria corticata

(Ragunathan et al., 2019), Sargassum wightii (Deepika, 2019) and Gracillaria dura

(Sumayya and Murugan, 2019). Each one of these identified secondary metabolites are

S. No Metal chelating

(mg EDTA/g

sample)

TAA

(μmol/g extract)

1 30.62 mg/g 2863.19

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known to have a wide range of biological activities including anti-microbial, anti-oxidant,

anti-spasmodic, anti-tumour activity, anti-fungal, nematicidal, anti-androgenic, 5-Alpha

reductase inhibitor, Pesticide, Lubricant, Haemolytic, anti-cancer, anti-diabetic, anti-

inflammatory, Hepatoprotective, Muscle weakness, Drowsiness, Sleep disturbance, anaemia,

hyperthermic and hepatitis. Pentadecanoic acid that have reported to possess high

antibacterial, antifungal and antioxidant properties (Sermakkani and Thangapandian, 2012;

Santhoshkumar et al., 2014). Methyl esters such as n-hexadecanoic acid (Palmitic acid),

eicosanoic acid (Arachidic acid), Lucenin 2, Benzoic acid and pentadecanoic acid have

shown potential to inhibit various bacterial pathogens such as Escherichia coli, Klebsiella

pneumoniae, Pseudomonas aeruginosa, that causes various life-threatening skin infections

were inhibited by methyl esters. Free saturated fatty acids like n-hexadecanoic acid,

eicosanoic acid, oleic acid and pentadecanoic acid have shown immense potential to inhibit

the growth of gram-positive bacteria. Whereas, the fatty acids have shown potential to inhibit

the methicillin-resistant bacteria without any side effects. 1H-Cyclopropa[3,4]benz[1,2-

e]azulene-4a,5,7b,9,9a(1 aH)-pentol has shown inhibition against leishmanias by producing

Reactive Oxygen Species (ROS) and free radicals and causing the death of the parasite

(Ragunathan, et al., 2019).

The n-hexadecanoic acid also has antifungal property which induced the ROS

resulting in the increased oxidative stress inside the cell and ultimately leading the

mitochondrial disfunction and activating the pro-apoptotic factors. Hexadecanal, also known

as palmitaldehyde have shown to resist the growth of pathogenic microorganisms such as

Bacillus cereus and Pseudomonas aeruginosa and also has antioxidant properties in the

DPPH. GC-MS analysis compounds have anti-tumour, anti-oxidant, anti-bacterial, anti-

pesticidal and anti-nematicidal activity were identified and reported by Jenifer and

Balakrishnan, (2015). The results of the present study consented on that medicinally

important compounds present which was reported by Akpuaka et al., (2013). In the previous

study, Nadathur et al., (1996) identified that the anti-mutagenic compound in the GC-MS

analysis of seaweed. Methanol extract of U. flexuosa, GC-MS results were coincided with

Ragunathan, et al., (2019) and Purushoth et al., (2013).

.

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Figure 2. Chromatogram of the methanol extract of Green algae

Figure 3. Mass spectrum of the compound Hexadecanoic acid, methyl ester.

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Figure 4. Mass spectrum of the compound 13-Docosenamide, (Z)

Figure 5. Mass spectrum of the compound Eicosane

Figure 6. Mass spectrum of the compound Cholest-5-en-3-ol, 24-propylidene-, (3á)-

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Figure 7. Mass spectrum of the compound 9-Octadecenoic acid (Z)-, methyl ester

(CAS)

Figure 8. Mass spectrum of the compound Z-(13,14-Epoxy) tetradec-11-en-1-ol acetate

Figure 9. Mass spectrum of the compound7-Methyl-Z-tetradecen-1-ol acetate.

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Table 3: Identification of chemical compounds by using Gas Chromatography and Mass Spectroscopy analysis of methanolic extracts of

Ulva flexuosa.

S. No Compound name Retention

Time (RT)

Peak Area

(%)

Molecular formula

(MF)

Molecular weight

(MW)

1 N-BZ-2amino cinnamate 6.24 1.67 C17H17NO2 267

2 Dotriacontane (CAS) 7.61 0.92 C32H66 450

3 Tetradecane (CAS) 10.50 1.15 C14H30 198

4 2,2,3,3,4,4 hexadeuterooctadecanal 14.45 0.62 C18H30D6O 268

5 Z-(13,14-Epoxy) tetradec-11-en-1-ol acetate 16.10 0.54 C16H28O3 268

6 Eicosane 16.51 10.24 C20H42 282

7 Methyl tetradecanoate 17.04 3.30 C15H30O2 242

8 Hexadecanoic acid (CAS) 18.34 2.23 C16H32O2 256

9 2-Pentadecanone, 6,10,14-trimethyl- (CAS) 19.50 1.33 C18H36O 268

10 2,3-Dihydroxypropyl elaidate 20.69 0.72 C21H40O4 356

11 Hexadecanoic acid, methyl ester 21.07 29.10 C17H34O2 270

12 3,6-bis(t-Butyl) fluorenone 21.46 3.13 C21H24O 292

13 9-Octadecenoic acid (Z)-, methyl ester (CAS) 24.38 8.07 C19H36O2 296

14 7-Methyl-Z-tetradecen-1-ol acetate 24.63 0.54 C17H32O2 268

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15 Tetraneurin - A – diol 27.33 1.23 C15H20O5 280

16 Hexadecanoic acid, 2,3-dihydroxypropyl ester (CAS)

29.25 0.69 C19H38O4 330

17 9,12,15-Octadecatrienoic acid,2,3-bis[(trimethylsilyl) oxy] propyl ester, (Z, Z, Z)

30.84 0.65 C27H52O4Si2 496

18 Hexa-t-butyl selenatrisiletane 31.08 0.99 C24H54SeSi3 506

19 13-Docosenamide, (Z) 34.46 10.27 C22H43NO 337

20 Lucenin 2 35.79 1.43 C27H30O16 610

21 5,7,9(11)-Androstatriene, 3-hydroxy-17-oxo- 38.19 1.35 C19H24O2 284

22 Cholest-5-en-3-ol, 24-propylidene-, (3á)- 39.09 9.00 C30H50O 426

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Table 4: Structure and biological significance of green alga Ulva flexuosa.

S.

No

Compound name Nature of the

compound

Structure Biological uses

1 N-BZ-2amino cinnamate Cinnamic acid - No activity reported.

2 Dotriacontane (CAS) Alkanes acyclic branched

hydrocarbon

Antimicrobial, antioxidant, antispasmodic

3 Tetradecane (CAS) Methyl alkanes

Antibacterial

4 2,2,3,3,4,4 hexadeuterooctadecanal

Steroldehyde

FALDH activity in patients suspected of having Sjogren-Larsson syndrome.

5 Z-(13,14-Epoxy) tetradec-11-en-1-ol acetate

Triterpenic acid

Antioxidant, Haemolytic.

6 Eicosane Acyclic Alkanes

Antitumour activity against the human gastric SGC-7901 cell line.

7 Methyl tetradecanoate Methyl ester

Antifungal, Antibacterial, Nematicidal

8 Hexadecanoic acid (CAS) Hydroxy

Palmitic acid

Antioxidant, Hypochloesterolemic, Nematicide, Pesticide, Lubricant, Antiandrogenic, Haemolytic, 5-Alpha reductase inhibitor.

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9 2-Pentadecanone, 6,10,14-trimethyl- (CAS)

Dihydroxyphenyl

Allelopathic, Antibacterial

10 2,3-Dihydroxypropyl elaidate

Campesteryl

No activity reported.

11 Hexadecanoic acid, methyl ester

Fatty acid Ester

Antifungal, Antioxidant, hypocholesterolemic nematicide, pesticide, antiandrogenic flavour, haemolytic, 5-Alpha reductase inhibitor, potent antimicrobial activity. [

12 3,6-bis(t-Butyl) fluorenone Ethyl phenol

No activity reported.

13 9-Octadecenoic acid (Z)-, methyl ester (CAS)

Fatty acid methyl

ester

Antioxidant, Hypochloesterolemic, Nematicide, Pesticide, Lubricant, Antiandrogenic, Haemolytic, 5-Alpha reductase inhibitor.

14 7-Methyl-Z-tetradecen-1-ol acetate

Acetate ester

Hepatoprotective, Anti-histaminic, Antieczemic, Hypocholesterolemic

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15 Tetraneurin - A – diol Acetate - Anticancer,

anti-inflammatory, Hepatoprotective

16 Hexadecanoic acid, 2,3-dihydroxypropyl ester (CAS)

Fatty acid, methy

ester

Anti-oxidant, Muscle weakness, Drowsiness, Sleep disturbance, Anaemia, Hyperthermic, Hepatitis

17 9,12,15-Octadecatrienoic acid,2,3-bis[(trimethylsilyl) oxy] propyl ester, (Z, Z, Z)

Oleic acid

No activity reported

18 Hexa-t-butyl selenatrisiletane

Fatty acid

Muscle weakness, Drowsiness, Sleep disturbance, Anaemia, Hyperthermic, Hepatitis.

19 13-Docosenamide, (Z) Alkadienes

Muscle weakness, Drowsiness, Sleep disturbance, Anaemia, Hyperthermic, Hepatitis

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20 Lucenin 2

Glucoside

Dermatitis, mouthwash, vaginal-douche-and veterinary activities

21 5,7,9(11)-Androstatriene, 3-hydroxy-17-oxo-

Steroid

Anti-microbial activity, it is used to control estrogen synthesis (Adkins-Regan et al., 2006)

22 Cholest-5-en-3-ol, 24-propylidene-, (3á)-

Steroid

It serves as a precursor for the biosynthesis of steroid hormones, bile acids, and vitamin D (Hanukoglu et al., 1992)

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IN VITRO ANTIBACTERIAL ACTIVITY

In the present study, the antibacterial activities of the methanolic extracts of green

algae Ulva flexuosa seaweed to investigated against human pathogenic bacteria such as

Streptococcus pyogenes, Staphylococcus aureus (Gram positive), Escherichia coli and

Klebsiella pneumonia (Gram negative). The plants species Ulva flexuosa showed very high

inhibitory activity against Staphylococcus aureus (26.02±1.2) followed by Streptococcus

pyogenes (22.54±1.0), Escherichia coli (20.40±0.7) and Klebsiella pneumoniae (19.31±0.7)

than standard drug Erythromycin. The zone of inhibition measured and summarized in

(Table 5).

In the present study, methanol solvent extract of Ulva flexuosa displayed highest

inhibitory activity against the test pathogens as compared to standard. These results are

parallel to the highest content of total phenol and total flavonoids in extract, thus the

antibacterial activity of Ulva flexuosa might be attributed to their phenolic and flavonoid

content. In this concern, a positive relationship between antimicrobial activity potential and

amount of phenolic compounds of the crude extracts was reported (Mohamed et al., 2015).

.

Figure: 10 a] Staphylococcus aureus b] Streptococcus pyogenes c] Escherichia coli d] Klebsiella pneumoniae

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Table 5. In vitro antibacterial activity of methanolic extracts of Ulva flexuosa

CONCLUSION

From the Above results concluded that the methanolic extracts of Ulva flexuosa having rich amount of phenol and flavonoids content; In addition, Gas chromatography and mass spectrum analysis totally twenty-two biological active compounds were identified. Moreover, the results proved that the selected seaweeds have been taken as a feed for animals and also used for agricultural purposes. The species Ulva flexuosa, having several biological chemical compounds used as source of antibiotic, antioxidant, anti-inflammatory, anti-cancer properties. Hence, it has great potential in prebiotic food resource for animal and human health.

CONFLICT OF INTEREST: There is no Conflict of Interest in this Article.

ACKNOWLEDGEMENTS We thankful to the Research Department of Zoology, Kongunadu Arts and Science

College, Coimbatore for providing the necessary lab facilities and support for this paperwork.

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Ulva flexuosa Erythromycin

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