Pharmacogn J.

2022; 14(3): 641-649

A Multifaceted Journal in the field of Natural Products and Pharmacognosy
Research Article
www.phcogj.com

Phytochemical Screening, Gc-Ms Analysis and Antioxidant

Activity of Marine Algae Obtained from Coastal Andhra Pradesh,
India
Swathi Priya K1,*, Rajasekaran S2

ABSTRACT
Swathi Priya K1,*, Rajasekaran S2 Introduction: We have seen much research on novel compounds obtained from natural sources, but in
the recent era, it's seen that there is tremendous scope for investigation on marine sources. In the past,
1
Research Scholar, Department of Pharmacy, due to a lack of technology, there was less knowledge about the treasure hidden in marine sources.
Bhagwant University, Sikar Rd, Ajmer, Because of the latest technology, it's been an easy way of collecting marine sources and investigating
Rajasthan, INDIA. them. Marine organisms are the source of highly bioactive secondary metabolites, which might help
2
Department of pharmacology, Bhagwant develop new pharmaceutical agents. Marine algae are classified into two main groups, i.e., microalgae
University, Sikar Rd, Ajmer, Rajasthan, INDIA. and macroalgae. Microalgae include blue-green algae, bacillariophyte and dinoflagellates, whereas
macroalgae, mostly known as seaweed, are divided into green, brown and red algae. Material and
Correspondence methods: For the present study, a green macroalga named spongomorpha indica has been selected,
Swathi Priya K
and it’s been investigated for its physico chemical parameters, phytochemical study, GC-MS analysis and
antioxidant activity. This investigation was performed to know whether this seaweed has the potential
Research Scholar, Department of
Pharmacy, Bhagwant University, Sikar Rd,
requirements for further research to see whether it's useful for healthcare. Results: The physicochemical
Ajmer, Rajasthan, INDIA. parameters results obtained were in accordance with the who guidelines and the phytochemical tests
revealed the presence of potent active constituents like alkaloids, steroids, tannins, flavonoids because
E-mail: [email protected]
of which the study was further extended to GCMS analysis where seven components were identified
History among which has the highest peak and has the highest molecular weight. And finally it was tested for
• Submission Date: 13-04-2022; antioxidant activity using four different models all the results showed best antioxidant activity, among
which superoxide scavenging activity showed the best results. Conclusion: basing upon the results
• Review completed: 02-05-2022;
obtained it was proved that spongomorpha indica have potent active constituents which shows great
• Accepted Date: 10-05-2022. antioxidant effect and hence the study is further proceeded to check target related activity so that it can
DOI : 10.5530/pj.2022.14.83 produce best results in curing a particular disease.
Key words: Spongomorpha indica, Superoxide, DPPH, GCMS analysis, Physicochemical.
Article Available online
http://www.phcogj.com/v14/i3
Copyright INTRODUCTION in many parts of the world. They have been screened
© 2022 Phcogj.Com. This is an open-
extensively to isolate life-saving drugs or biologically
In today's economic life of humans and ecosystems, active components worldwide.6 Traditional and
access article distributed under the terms
of the Creative Commons Attribution 4.0 the role of seaweeds is best known. Algae, mainly complementary medicine used active compounds
International license. the macroalgae commonly known as seaweeds, produced by marine organisms. Many varieties of
are used as food, animal fodder, fertilizers, raw
marine algae reported active compounds which
materials in the production of industrial products,
can cure diseases. Due to the fewer side effects
and as natural feed for economically critical aquatic
caused by drugs made of natural origin, most of the
species has received attention in Thailand and in
population prefers using natural-origin remedies to
many other countries around the world such as
cure diseases.7 Many pharmacological studies have
Japan,1 China,2 and other Asian countries (e.g.,
Korea, Philippines, India).3 Seaweeds serve as an reported that the chemical compounds produced by
essential source of bioactive natural substances.4,5 marine algae have different biological activities such
as anti-HIV, anticancer, antimutagenic, scavenging
Seaweeds are classified into three main categories free radicals and anti-inflammatory.8,9 Previous
based on their pigmentation: Phaeophyta, studies on marine microalgae reported that over
Rhodophyta and Chlorophyta. Phaeophyta or 15,000 compounds had been isolated, including
brown seaweeds are brown macroalgae due to sterols, fatty acids, terpenes, phenolic compounds,
the presence of the carotenoid fucoxanthin. enzymes, alkaloids, flavonoids and polysaccharides.
The primary polysaccharides present include Recently it was reported that marine algae are a
alginates, laminarins, fucans and cellulose.
source of antioxidant compounds with free radical
Rhodophyta or red seaweeds are red-pigmented
scavenging activity.10
macroalgae, whereas Chlorophyta, or green
seaweeds, are dominated by chlorophyll a and b, The preliminary screening of Spongomorpha
with ulvan the major polysaccharide component indica was performed in the present study to know
present in it. Seaweeds are the best sources of the phytochemical constituents present. GC-MS
bioactive compounds with cytostatic, antiviral, analysis was done to identify and analyze different
antihelminthic, antifungal, antibacterial and many components present in the extract of Spongomorpha
more biological activities. Seaweeds are natural indica. Also, the study aimed to investigate the
renewable sources used as food, feed and fertilizer antioxidant potentials of Spongomorpha indica.

Cite this article: Swathi Priya K, Rajasekaran S. Phytochemical Screening, Gc-Ms Analysis

Phcogj.com
and Antioxidant Activity of Marine Algae Obtained from Coastal Andhra Pradesh, India.
Pharmacogn J. 2022;14(3): 641-649.

641 Pharmacognosy Journal, Vol 14, Issue 3, May -June, 2022

 Swathi Priya K, et al.: Phytochemical Screening, Gc-Ms Analysis and Antioxidant Activity of Marine Algae Obtained from Coastal Andhra Pradesh, India

MATERIAL AND METHODS were calculated by using the following formula:

Selection and collection of Spongomorpha indica Weight of ash residue

Past literature study reveals that not much work was done on this Water-soluble ash% = ------------------------------ x 100
seaweed. Still, potentially active constituents present in this seaweed Weight of dry powder
can provide scientific proof of specific activity towards any disease. Hence,
it's been collected from Visakhapatnam coastal area during the low tidal
conditions. The collected sample was submitted to the herbarium of the Determination of acid insoluble ash
botany department of Andhra University and was authenticated.
The total ash for 5 minutes was boiled with 25 ml of dilute hydrochloric
Preparation of extract acid, in a Gooch crucible or on an ash-less filter paper. The insoluble
matter was collected, washed with hot water, ignited, and weighed.
The collected crude sample was washed thoroughly, shade dried and
the extract was prepared by the Soxhlet extraction method. Initially, 10 The percentage of acid-insoluble ash was calculated with reference to
kgs of the crude sample were kept for maceration separately with three the air-dried drug. The weights obtained were calculated by using the
solvents, i.e., ethyl acetate, hexane and methanol: water (70:30) for About following formula:
48 hours. The solvent was collected, evaporated under steam distillation,
and concentrated until a thick, greasy like consistency mass was obtained. Weight of acid-insoluble ash
The three extracts were separately tested for active constituents. The dried Acid insoluble ash % = ------------------------------------- x 100
extract was kept in the refrigerator at 4°C till future use. Weight of dry powder

Physicochemical parameters11-20
The physicochemical parameters used in this study are to analyze the Determination of sulphated ash
extract's quality and purity. For example, the Ash value helps determine
The silica crucible was heated until it turned red for 10minutes, allowed
the authenticity and purity of drugs, and also these values are also
to cool in desiccators, and weighed.1g of substance was accurately
crucial for quantitative standards. The extracts were evaluated for
weighed and was transferred to the crucible. At first, it was ignited
physicochemical parameters like a loss on drying, total ash, insoluble
gently until the whole substance was thoroughly charred. Then the
acid ash, water-insoluble ash and different extractive values according
residue was cooled and mixed with 1 ml concentrated sulfuric acid,
to the official methods described in the Indian Pharmacopeia and
heated gently until white fumes no longer evolved, and ignited at 800°c
WHO guidelines regarding quality control methods for medicinal
=/- 25 until all black had disappeared.
plant materials. The following physicochemical parameters evaluated
studied plant materials were given below: The ignition was conducted in a place isolated from air currents. The
crucible was cooled, and to this, a few drops of concentrated sulfuric
Determination of ash values acid was added and heated. It was Ignited as before, allowed to cool
To determine the quality and purity of crude drugs, especially in powder and weighed. Repeated the process until two successive considers did
form Ash values are very helpful. The objective of incinerating the not differ by more than 0.5mg. Calculate the percentage of sulfated
seaweed is to remove all traces of organic matter, which may otherwise ash with reference to the air-dried drug. The weights obtained were
interfere in an analytical determination. On incineration, crude drugs calculated by using the following formula:
typically leave ash, usually consisting of carbonates, phosphates, and
silicates of sodium, potassium, calcium and magnesium. W3-w1
Sulphated ash% = ------------------ x 100
Determination of total ash W
About 3 grams (accurately weighed) of seaweed powder was taken in a
silica crucible which was incinerated and considered previously. It was
Since w1 = crucible weight
incinerated by gradually increasing the heat, not exceeding red heat
(450°c), until it was free from carbon, cooled down, and weighed. The W2 = crucible wt. + sample
percentage of ash has been calculated concerning the air-dried powder.
The procedure was repeated five times to get constant weight. The W3 = final wt. of crucible + sample
obtained weights were calculated by using the following formula: W = sample taken

Weight of ash Loss on drying

Ash% = ---------------------------------- x 100 The loss on the drying test measures the amount of water and volatile
Weight of dry powder matter present in a sample when the sample is dried under specified
conditions. The weighing bottle was dried for about 30min, cooled in
the desiccators, and weighed again accurately. 5gms of the sample was
Determination of water-soluble ash taken into a weighing bottle, and the sample was spread as a thin layer
of less than 5mm and weighed accurately and placed in oven-dried at a
Boil the total ash obtained in the above process for 5 minutes with 25ml temperature of about 10 degrees for 1/2hour and cooled and weighed
of water. In a Gooch crucible or in an as-less filter paper, insoluble until a constant weight was obtained. The weights obtained were [
matter was collected, washed shed with hot water, and incinerated to calculated by using the following formula:
constant weight at a low temperature. The weight of insoluble matter
was removed from the weight of the ash; the difference in weight Initial weight – final weight
represents the water-soluble ash. The percentage of water-soluble ash Loss on drying % = --------------------------------------------------- x 100
was calculated concerning the air-dried drug. The weights obtained Final weight

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 Swathi Priya K, et al.: Phytochemical Screening, Gc-Ms Analysis and Antioxidant Activity of Marine Algae Obtained from Coastal Andhra Pradesh, India

Swelling index was removed and transferred to the ashing dish (pre-weighed dish
W1). The residue was dried for 2 hours at 130 ±2°C. Cool the dish in
Under specified conditions, the swelling index is the volume in ml desiccators and weigh (W2). Ignite for 30min at 600±15°C. Cool in
taken up by the swelling of 1gm of herbal material. As specified, its desiccators and reweigh (W3). The weights obtained were calculated
determination was based on the addition of water or a swelling agent. by using the following formula:
About 1gm of the powdered drug was taken in a glass stoppered
measuring cylinder and added water of about 25ml and shaken
(W2-W1) - (W3-W1)
repeatedly for about 1hour and allowed to stand it for 24hours and
Crude fiber content % = ------------------------------------ X 100
volume in ml is read is measured. The procedure was repeated two
Weight of the drug sample
times. The weights obtained were calculated by using the following
formula:
Since W1= Empty China dish weight.
Final weight – initial weight
Swelling index%= ------------------------------------------- x 100 W2= Powder weight after oven-dried and cooled
Final weight W3= Powder weight after igniting and cooled

Foreign organic matter

Foaming index Weigh 200gms crude sample accurately and evenly spread into a thin
The ability to produce foam of an aqueous decoction of herbal material layer. The sample was inspected with the unaided eye or used a 6x
and extracts is measured using a foaming index. About 1gram of the lens, and the foreign organic matter was removed manually altogether
powdered drug was taken into a 500ml conical flask, and 100ml of as much as possible. Finally, it was Weighed, and the weight of the
boiling water was added to it and maintained at moderate boiling for drug taken determined the percentage of foreign organic matter. The
30 min; collected and filtered into a 100ml conical flask. The decoction maximum samples have to be used for coarse or bulky drugs.
was taken into ten stoppered test tubes add it in successive portions of
1ml, 2ml,3ml,…….10ml. The volume was adjusted to 10ml, and the PHYTOCHEMICAL SCREENING OF SPONGO-
test tubes were stoppered and shaken for 15 seconds lengthwise, two MORPHA INDICA
shakes per second, and allowed to stand for 15 mins and the length of
foam was measured. Preliminary phytochemical analysis was carried out for three different
solvent-based extracts of Spongomorpha Indica i.e., hexane, ethyl acetate
The foaming index was calculated using the following formula: and hydro methanolic extracts by applying standard procedures.21-24
Foaming index = 1000/a Detection of alkaloids
Where a= the volume in ml of the decoction used to prepare the tubes, The three different extracts were dissolved individually in dilute
dilution foaming to a height of 1cm was observed. hydrochloric acid and filtered. The filtrate was used to test the presence
Extractive values of alkaloids by the following tests.

5gms of an air-dried coarse powder of drug macerated with 40 ml of Mayer's test

solvents (hexane, ethyl acetate, methanol and methanol: distilled water
Filtrates were treated with Mayer's reagent. The Mayer's reagent is
(70:30) in a glass stoppered conical flask with frequent shaking for
prepared by dissolving Mercuric chloride (1.358g) in 60ml of water
6hours and then allowed to stand for two days, then filtered carefully,
taking care against loss of solvent. The filtrate was evaporated in a silica and potassium iodide (5g) in 10ml of water. The two solutions were
crucible to dryness on a water bath and then dried at 105 for 6 hours, mixed and the volume was made upto 100ml of water. The formation of
cooled in a desiccator for 30mins, and weighed without delay. The yellow color on the addition of Mayers reagent to filtrates determines
weights obtained were calculated by using the following formula: the presence of alkaloids.

Wagner's test
Residue weight Filtrates were treated with Wagner's reagent. The formation of brown/
Extractive values % = --------------------------- x 100 reddish brown precipitate indicates the presence of alkaloids. Wagner's
Weight of the sample
reagent was prepared by dissolving iodine (1.2g) and potassium iodide
(2g) in 5ml of water and the volume is made up to 100ml with distilled
water.
Crude fiber content
2grams of ground material were extracted with ether or petroleum Detection of flavonoids
ether to remove fat (initial boiling temperature 35-38°c and final Lead acetate test
temperature 52°c). An extraction might be omitted if the fat content
was below 1%. After extraction with ether,2gms of dried material Extracts were treated with a few drops of lead acetate solution. The
was boiled with 200ml of sulphuric acid for 30mins with bumping formation of a yellow color precipitate indicates the presence of
chips. The whole liquid was filtered through muslin and washed with flavonoids.
boiling water until the washings were no longer acidic. 200ml of
H2SO4 test
sodium hydroxide solution was added and heated for 30min. Filtered
through muslin cloth again and washed with 25ml of boiling 1.25% Extracts were treated with a few drops of H2SO4. The formation of
H2SO4, three 50ml portions of water, and 25ml alcohol. The residue orange color is a indication for the presence of flavonoids.

643 Pharmacognosy Journal, Vol 14, Issue 3, May -June, 2022

 Swathi Priya K, et al.: Phytochemical Screening, Gc-Ms Analysis and Antioxidant Activity of Marine Algae Obtained from Coastal Andhra Pradesh, India

Detection of steroids GC-MS analysis

Liebermann- Burchard test The components present in the extract were separated using Helium as
carrier gas at a constant flow of 1 ml/min, Clarus 680 GC was used to
2ml of acetic anhydride was added to 0.5g of the prepared extracts, each employ a fused silica column packed with Elite-5MS (5% biphenyl 95%
with 2ml of H2SO4. if their is a color change from violet to blue or dimethylpolysiloxane, 30 m × 0.25 mm ID × 250μm df) and During
green in some samples is a indication for the presence of steroids the chromatographic run, the injector temperature was set at 260°C.
The 1μL of extract sample was injected into the instrument; the oven
Detection of terpenoids
temperature was as fixed to 60 °C (2 min); followed by 300 °C at the
Salkowski's test rate of 10 °C min−1; and 300 °C, where it was held for 6 min. The mass
detector conditions were: transfer line temperature 240 °C; ion source
Chloroform and concentrated sulphuric acid (3ml) were carefully temperature 240 °C; ionization mode electron impact at 70 eV, a scan
added to 0.2g of the extract of the whole sample to form a layer. the time of 0.2 sec, and scan interval of 0.1 sec—the fragments from 40
presence of terpenoids is determined by a reddish-brown coloration to 600 Da. The spectrums of the components were compared with the
of the inner face. database of the spectrum of known compounds stored in the GC-MS
Detection of anthroquinones NIST (2008) library. Measurement of peak areas and data processing
were carried out by Turbo-Mass OCPTVS-Demo S.P.L. software.
Borntrager's test
Antioxidant activity
0.2g of the extract was boiled with 10% Hydrochloric acid for a few
minutes in a water bath. It was filtered and allowed to cool. the same Antioxidants are compounds that inhibit oxidation. Free radicals can
volume of chloroform was added to the filtrate. A few drops of 10% be produced through a chemical reaction named Oxidation, thereby
ammonia was added to the mixture and heated. Anthraquinones leading to chain reactions that may damage the cells of an organism.
presence can be indicated by the formation of pink color. Well-known antioxidants include various enzymes and other
substances, such as beta carotene, vitamin C and vitamin E, which can
Detection of phenols counter the damaging effects of oxidation. Antioxidation terminates
these chain reactions, such as thiols or ascorbic acid (vitamin C). To
Ferric chloride test
balance the oxidative stress, animals and plants maintain complex
The formation of bluish-black color when the extracts are treated with systems of overlapping antioxidants such as enzymes (e.g., superoxide
5% ferric chloride solution indicates the presence of phenol. dismutase and catalase) and glutathione, produced internally in a
human system or the dietary antioxidants such as vitamin C and
Lead acetate test Vitamin E. Antioxidants may reduce the risk of cancer. The progression
The formation of a yellow color precipitate when the extracts were of age-related macular degeneration is slowed down with the help of
treated with few drops of lead acetate solution indicates the presence antioxidants. This process is called Antioxidant activity.
of phenolic compounds.
Reducing power assay25
Detection of saponins The above sample, including extract with Ascorbic acid solutions, was
Froth test spiked with 2.5ml of phosphate buffer (0.2 M, pH 6.6) and 2.5ml of
1% potassium ferricyanide. The mixture was kept in a 50°C water bath
Formation of frothing (appearance of creamy stable and persistent tiny for 20min. The resulting solution was then cooled rapidly, spiked with
bubbles) the extract was shaken with 5ml of distilled water appearance 2.5ml of 10% trichloroacetic acid, and centrifuged at 3000rpm for 10
of tiny bubbles indicates the presence of saponins. min. The supernatant liquid (5ml) was then mixed with 5ml of distilled
water and 1ml of 0.1% ferric chloride. The absorbance was detected at
Detection of tannins 700nm after reaction for 10min. The higher the absorbance represents,
Ferric chloride test the more substantial the reducing power. The reducing power assay
was expressed as ascorbic acid equivalent per gram of dry weight basis.
A small quantity of extract was mixed with water and heated in a water
bath. The mixture was filtered to which 0.1% ferric chloride was added. DPPH activity26,27
A dark green color formation indicates the presence of tannins.
DPPH radical scavenging activity was carried out by adding 1.0 ml of
Detection of carbohydrates 100.0 μM DPPH solution in methanol; an equal volume of the sample
in methanol of different concentrations was added and incubated in
Fehling's test the dark for 30 minutes. It was observed for colour change change
0.2gm filtrate from each extract was boiled in a water bath with 0.2ml in terms of absorbance using a spectrophotometer at 514 nm. To the
of Fehling solutions A and B. The presence of sugars can be determined control tube, 1.0 ml of methanol instead of the test sample was added.
by the indication of red precipitation. Fehling's solution A is prepared The different concentration of ascorbic acid was used as reference
by Copper sulphate (34.66g) dissolved in distilled water and made compound. The percentage of inhibition was calculated from the
up to 500ml using distilled water. Fehling' solution B is prepared equation.
by Potassium sodium tartrate (173g) and sodium hydroxide (50g)
[(Absorbance of control - Absorbance of test)/ Absorbance of control)]
dissolved in water and made up to 500ml.
×100. The IC50 value was calculated using Graph pad prism 5.0.
Detection of oils and resins
Superoxide radical scavenging activity28
Spot test
The superoxide radical scavenging activity of the test sample was studied
Apply test on a filter paper. If it develops a transparent appearance like using the method of Lee with slight modifications. Superoxide radicals
a stain on the filter paper, indicates the presence of oils and resins. are generated in phenazine methosulphate (PMS) - (Nicotinamide

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 Swathi Priya K, et al.: Phytochemical Screening, Gc-Ms Analysis and Antioxidant Activity of Marine Algae Obtained from Coastal Andhra Pradesh, India

adenine dinucleotide (NADH) systems by oxidation of NADH and phosphate buffer (pH 7.4, 0.1 M) was mixed with different concentrations
assayed by the reduction of Nitro Blue Tetrazolium (NBT). 200.0 μl of the extract (100 – 500 microgram/ml in phosphate buffer (pH 7.4, 0.1
of test samples of different concentrations were taken in a series of test M). The tubes were incubated at 250C for two hours. At the end of the
tubes. Superoxide radical was generated by 1.0 ml of Tris-HCl buffer second hour, 1.5 ml of the reaction mixture was removed and diluted
(16.0 mM, pH-8.0), 1.0 ml of NBT (50.0 μΜ),1.0 ml NADH (78.0 μΜ) with 1.5 ml of Greiss reagent (1% sulphanilamide, 2% o-phosphoric
solution and 1.0 ml of PMS (10 μM). The reaction mixture was incubated acid, 0.1% of naphthyl ethylenediamine dihydrochloride) The
at 25°C for 5 min and the absorbance at 560 nm was measured. A absorbance of the chromophore formed during diazotization of the
control tube containing Tris-HCl buffer was also processed in the same nitrite with sulphanilamide and subsequent coupling with naphthyl
way without a test sample. Different concentrations of ascorbic acid ethylene diamine dihydrochloride was measured at 546 nm. The
were used as a reference compound. Control tube contains all chemicals except plant extract.

Nitric oxide radical scavenging activity29 Evaluation of total antioxidant capacity of the extract30
Nitric oxide radical scavenging activity was measured by Phosphomolybdenum method was used to determine the total
spectrophotometry method. 1 ml of Sodium nitroprusside (5 mmol) in antioxidant capacity of the selected extract. The main mechanism

Table 1: Physicochemical parameters results.

Values obtained
S.No Parameters
(% w/w)
1 Total ash 38.33
2 Water soluble ash 9.8
3 Acid insoluble ash 14
4 Sulphated ash 13.59
5 Loss on drying 11.58
6 Swelling index 11.11
7 Foaming index <100
8 Crude fiber content 1.5
9 Foreign organic matter 1

Table 2: Extractive values results.

Values obtained
S.No Extractive values
(% w/w)
1 Hexane soluble extractive values 0.8
2 Acetone soluble extractive values 2.4
3 Ethyl acetate soluble extractive values 1.6
4 Methanol soluble extractive value 9.6
5 Methanol : distilled water (70:30) soluble extractive value 13.6

Table 3: Phytochemical screening results.

Extracts
Phytochemical tests Observations Hydroalcoholic
Hexane Ethyl acetate (70%methanol and 30%
water)
Alkaloids
Mayer’s test Cream color
_ _ +
Wagner’s test Reddish brown solution/ precipitate
Flavonoids
Lead acetate test Yellow orange
_ + +
H2SO4 test Reddish brown/ orange color precipitate
Steroids
Liebermann-buchard test Violet to blue or green color formation. + + +
Terpenoids
Salkowski test Reddish brown precipitate + + +
Anthroquinones
Borntragers test Pink colour - - +
Phenols
Ferric chloride test Deep blue to black colour formation
_ + +
Lead acetate test White precipitate
Saponin Stable persistent - + -
Tannin Brownish green/ blue black - + +
Carbohydrates Yellow/brownish/blue/green color + + +
Oil and resin Filter paper test + + +
Gums and mucilage + + +

645 Pharmacognosy Journal, Vol 14, Issue 3, May -June, 2022

 Swathi Priya K, et al.: Phytochemical Screening, Gc-Ms Analysis and Antioxidant Activity of Marine Algae Obtained from Coastal Andhra Pradesh, India

Table 4: GCMS results.

Si. No CAS Name of the Compound Molecular Formula Molecular Weight Retention Time Peak Area (%)
1,6-ANAHYDRO-3,4-DIDEOXY-.BETA.-D-
1 39682-48-9 C6H10O3 130 19.050 1.881
GLUCO-HEXOPYRANOSE
CYCLOPROPANEPENTANOIC ACID,2-
2 35305-79-9 C20H38O2 310 20.180 4.128
UNDECYL-,METHYL ESTER, TRANS-
3 900143-83-9 DECYL TRIFLUUOROACETATE C7H16O 116 20.411 5.049
4 90526-63-5 2,3-EPOXYHEXANOL C6H1202 116 20.631 7.615
5 13205-57-7 1-METHYLDODECYLAMINE C13H29N 199 21.011 32.780
6 646-30-0 NONDECANOIC ACID C19H38O2 298 22.126 47.006
7 41446-54-2 4-TRIDECENE,(Z)- C13H26 182 24.187 1.541

Table 5: Antioxidant results.

Reducing Power Superoxide Radical Nitricoxide radical
DPPH assay
Sl.No Concentration µg/ml assay Scavenging Assay Scavenging assay
% Inhibition IC50 % Inhibition IC50 % Inhibition IC50 % Inhibition IC50
1 50 22.94 40.03 22.65 38.10
2 100 49.25 49.87 34.32 51.21
76.72 93.52 162.96 109.83
3 150 70.13 67.21 47.12 55.65
4 200 82.98 81.34 51.02 64.98

Qualitative Report
File: C:\TurboMass\2021.PRO\Data\ME-(21ES-0604).raw
Acquired: 25-Oct-21 05:09:36 PM Printed: 27-Oct-21 03:36 PM
Description:
GC/MS Method: GC: METHOD-1.mth MS: METHOD-1.EXP Page 1 of 1
Sample ID: ME-(21ES-0604) Vial Number: 51

ME-(21ES-0604) Scan EI+

TIC
100
7.95e7
2.64

%
3.31
3.61
4.32
4.94
5.55
6.16

22.13
20.85
20.18

22.06

22.73
23.34
6.79

23.95
24.56
25.19
7.39

25.80
19.05

26.40
27.00
27.62
19.35
8.01

28.24
10.45

28.84
11.05
8.62

11.67

29.45
30.06
30.67
12.27

18.63
9.22

31.89
12.94

31.27
9.85

0 Time
5.00 10.00 15.00 20.00 25.00 30.00

# RT Scan Height Area Area % Norm %

1 19.050 3309 2,223,949 329,455.6 1.881 4.00

2 20.180 3535 6,671,416 723,288.2 4.128 8.78
3 20.411 3581 6,053,358 884,621.8 5.049 10.74
4 20.631 3625 6,747,062 1,334,127.5 7.615 16.20
5 21.011 3701 6,835,764 5,742,834.0 32.780 69.73
6 22.126 3924 6,125,240 8,235,257.0 47.006 100.00
7 24.187 4336 1,595,983 269,919.8 1.541 3.28

Inst() ACQUISITION PARAMETERS

Oven: Initial temp 60°C for 2.50 min, ramp 10°C/min to 300°C, hold 6 min, InjAauto=260°C, Volume=0 µL, Split=10:1, Carrier
Gas=He, Solvent Delay=2.50 min, Transfer Temp=150°C, Source Temp=150°C, Scan: 40 to 600Da, Column 30.0m x 250µm

, 25-Oct-2021 + 17:09:36
ME-(21ES-0604) NR+Sm (SG, 2x4) Scan EI+
100 TIC
7.66e7

3.39

20.97 22.20

0 Time
7.50 12.50 17.50 22.50 27.50

Figure 1

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 Swathi Priya K, et al.: Phytochemical Screening, Gc-Ms Analysis and Antioxidant Activity of Marine Algae Obtained from Coastal Andhra Pradesh, India

Totalantioxidantactivity –153
Figure 2

involved in this process is the reduction of Mo (VI) to Mo (V) by the The antioxidant activity of hydroalcoholic extract of spongomorpha
antioxidant compounds present in the extract and formation of a green indica was studies using the different models i.e; Reducing power
Mo (V). An aliquot of sample solution (0.1ml) containing a reducing assay, DPPH assay, Superoxide radical scavenging assay, Nitric oxide
species in DMSO was combined in an Eppendorf tube with 1ml of radical scavenging assay shown in Table 5 and Figure 2. In all the
reagent solution (0.6M Sulphuric acid, 28mM sodium phosphate, and four assays it is seen that the percentage inhibition of the extract was
4mM ammonium molybdate). The tubes were capped and incubated directly proportional to concentration of the extract. Spongomorpha
in a water bath at 95 °C for 90min. The samples were cooled to room indica showed a significant dose dependent reduction in case of DPPH
temperature, and the absorbance of each solution was measured at
radical in the DPPH assay model. The highest IC50 value was seen in
695nm. The total antioxidant capacity was expressed as mM equivalent
the superoxide radical scavenging activity. Total antioxidant activity of
of ascorbic acid.
hydroalcoholic extract of spongomorpha indica found to be 153.
RESULTS AND DISCUSSION
CONCLUSION
The physicochemical parameters and extractive values obtained were
according to the Indian pharmacopeia and WHO guidelines standards. The By thorough study of past literature about spongomorpha indica it was
table 1 and 2 shows all the results observed and recorded during the study. observed that these seaweeds are used as food in many countries and
is mainly used as a fodder in aquaculture. And there has been only
The phytochemical screening for the selected sample was done by few medicinal studies performed. In the present study, the extraction
using three different solvent systems i.e; hexane, ethyl acetate and process was done using three solvent systems i.e.; hexane, ethyl acetate
hydroalcoholic (hydro methanolic) mentioned in table 3. And it was
and methanol:water and extracts were obtained screening for active
observed that except saponins all other active compounds were available
constituents. Basing upon results obtained i.e.; containing tannins,
in hydro methanolic extract of spongomorpha indica compared to the
remaining two solvent derived extracts. From the results obtained we alkaloids, phenols, steroids etc the study was further proceeded with
can study that hydro methanolic extract produced more compounds hydro-methanolic extract. GCMS results revealed the presence of seven
compared to the other solvent based extracts showing the active compounds with different nature present in the hydro methanolic
compounds like alkaloids, phenols, steroids, tannins etc which are extract, it also showed significant antioxidant potential with the four
essential medicinal compounds. And hence, for further studies hydro different assays performed. Thus, further study is continued to check
methanolic extract was selected. the medicinal potentiality of this extract having important active
compounds which can be helpful treating targeted disease which could
The GCMS results shown in Table 4 and Figure 1 revealed the presence
of seven different compounds. The compounds exhibited have a wider be helpful for the mankind.
range in their nature. Nondecanoic acid was observed to exhibit
largest peak area of about 47.006% with retention time 22.126 whereas
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GRAPHICAL ABSTRACT

ABOUT AUTHORS

K. Swathi priya is a Research Scholar, Department of Pharmacy, Bhagwant University, Sikar Rd,
Ajmer, Rajasthan, INDIA.

Rajasekeran .S, Department of pharmacology, Bhagwant University, Sikar Road, Ajmer, Rajasthan,
INDIA.

Cite this article: Swathi Priya K, Rajasekaran S. Phytochemical Screening, Gc-Ms Analysis and Antioxidant Activity of Marine Algae
Obtained from Coastal Andhra Pradesh, India. Pharmacogn J. 2022;14(3): 641-649.

649 Pharmacognosy Journal, Vol 14, Issue 3, May -June, 2022