Preliminary Phytochemical Analysis of Selected Seagrasses

in Guiuan, E. Samar
Glenn Dioshua O. Altar
Guiuan National High School

Abstract
Seagrasses are angiosperms that lives in marine environment. They serve as nursery grounds, food source
and shelter for many sea organisms. Recent studies on seagrass phytochemistry demonstrated that they are
a significant wellspring of antioxidants, antibacterial agents, minerals and potential anticancer
compounds. On the other hand, plant produce phytochemicals to ensure themselves against ecological
dangers like predator bugs, contamination and diseases. Some activities of phytochemicals are; anti-
microbial, anti-diarrheal, etc. To date, little information exists about phytochemicals in E. acoroides, T.
hemprichii, and H. uninervis. Hence, this study aimed to determine the phytochemical constituents these
seagrasses have. Extracts were treated with reagent on each test with a color change as an indicator for
the presence of the phytochemical. After the tests, the phytochemical constituents were determined.
Introduction
Seagrass is an angiosperm that lives in marine environment1. Seagrasses have adjusted to develop in beach
front marine situations in both tropical and calm areas2. In general seagrasses inhabit the tidal and subtidal
zones of shallow and sheltered localities of seas, gulfs, bays, backwaters, lagoons and estuaries. They
typically lean toward sloppy, sandy, clayey and coral rubble substrate; however, they likewise develop on
rocks and in fissure3. They are known to be highly productive and serves as nursery grounds, food source
and shelter for many sea organisms1. They also reduce coastal erosions and provide habitat to both attached
and free-living organisms4. At least 12 seagrass species have been distinguished in the Philippines out of
the 50 species around the world5, while 9 species can be found in Guiuan, Eastern Samar6.

1
M. D. Richmond, A Guide to the Seashores of Eastern Africa: and the Western Indian Ocean Island
(Zanzíbar, Tanzania: SIDA. Department for Research Cooperation, SAREC, 1997).
2
R.G Coles, C.A Short, and F.T Short, Global Seagrass Research Methods (Amsterdam: Elsevier, 2002).
3
Tanaji G. Jagtap, Deepali S. Komarpant, and Rouchelle S. Rodrigues, “Status of a Seagrass Ecosystem: An
Ecologically Sensitive Wetland Habitat from India,” Wetlands 23, no. 1 (2003): pp. 161-170,
https://doi.org/10.1672/0277-5212(2003)023[0161:soasea]2.0.co;2.
4
Marten A. Hemminga and Carlos M. Duarte, Seagrass Ecology (Cambridge: Cambridge University Press, 2008).
5
Miguel D. Fortes, Seagrasses: A Resource Unknown in the ASEAN Region (Manila, Philippines: Published by the
International Center for Living Aquatic Resources Management on behalf of the Association of Southeast Asian
Nations/United States Coastal Resources Management Project, 1990).
6
Nonita Cabacaba et al., “Habitat Characteristic and Reproductive Biology of the Flat-Ribbed Scallop,
Decatopecten Radula (Linnaeus, 1758) on the Coastal Waters of Guiuan, Eastern Samar,” The Philippine Journal of
Fisheries 25, no. 1 (2018): pp. 107-117, https://doi.org/10.31398/tpjf/25.1.2017a0006.
 In addition, the significance of seagrasses has been archived in many coastal communities around
the world, including India7, Africa8, Canada9,10, Mexico11,12 and Sweden13. Some of these studies describe
the role of seagrasses in ecosystem function13, while others highlight their economic and traditional value,
such as the research of Wyllie-Echeverria and Cox13 concerning wild and commercial gathering of seagrass
Zostera marina by North American fishing communities during the early to mid-1900s for use as green
manure and insulating products. Seagrasses have likewise been read for their potential use in present day
drug7,14,15,16. Recent inquiries on seagrass phytochemistry has demonstrated its significance as wellspring
of antioxidants7, antibacterial agents15, minerals16 and potential anticancer compounds17.
Meanwhile, plant products have been part of phytomedicines for a very long time18. Plants produce
phytochemicals to ensure themselves against ecological dangers like predator bugs, contamination and
diseases19. This can be derived from barks, stems, roots, flowers, fruit, and seeds18. On the other hand, most
of the developing countries in the world still rely on herbal preparations for the treatment of some diseases
despite having conventional medicines20. Some activities of phytochemicals are; anti-microbial which are
present in phenols, flavonoids, tannins, coumarins, terpenoids, and alkaloids; anti-diarrheal which are

7
Rengasamy Ragupathi Raja Kannan, Rajasekaran Arumugam, and Perumal Anantharaman, “In Vitro Antioxidant
Activities of Ethanol Extract from Enhalus Acoroides (L.F.) Royle,” Asian Pacific Journal of Tropical Medicine 3,
no. 11 (2010): pp. 898-901, https://doi.org/10.1016/s1995-7645(10)60216-7.
8
D. Pillay, Gm Branch, Cl Griffiths, C Williams, and A Prinsloo. “Ecosystem Change in a South African Marine
Reserve (1960–2009): Role of Seagrass Loss and Anthropogenic Disturbance.” Marine Ecology Progress Series 415
(2010): 35–48. https://doi.org/10.3354/meps08733.
9
Sandy Wyllie-Echeverria, and Paul Alan Cox. “The Seagrass (Zostera Marina [Zosteraceae]) Industry of Nova
Scotia (1907–1960).” Economic Botany 53, no. 4 (1999): 419–26. https://doi.org/10.1007/bf02866721.
10
HV Kuhnlein, NJ Turner. “Traditional Plant Foods of Canadian Indigenous Peoples: Nutrition, Botany and Use.”
Choice Reviews Online 29, no. 11 (January 1992). https://doi.org/10.5860/choice.29-6359.
11
R Felger, and M. B. Moser. “Eelgrass (Zostera Marina L.) in the Gulf of California:” Science 181, no. 4097
(1973): 355–56. https://doi.org/10.1126/science.181.4097.355.
12
Eugene S. Hunn, “: People of the Desert and Sea: Ethnobotany of the Seri Indians. Richard Stephen Felger, Mary
Beck Moser.” American Anthropologist 88, no. 4 (1986): 980–81. https://doi.org/10.1525/aa.1986.88.4.02a00400.
13
Patrik Rönnbäck, Nils Kautsky, Leif Pihl, Max Troell, Tore Söderqvist, and Håkan Wennhage. “Ecosystem
Goods and Services from Swedish Coastal Habitats: Identification, Valuation, and Implications of Ecosystem
Shifts.” AMBIO: A Journal of the Human Environment 36, no. 7 (2007): 534–44.
https://doi.org/10.1579/00447447(2007)36[534:egasfs]2.0.co;2.
14
K. Alam, T. Agua, H. Maven, R. Taie, K. S. Rao, I. Burrows, M. E. Huber, and T. Rali. “Preliminary Screening of
Seaweeds, Seagrass and Lemongrass Oil from Papua New Guinea for Antimicrobial and Antifungal Activity.”
International Journal of Pharmacognosy 32, no. 4 (1994): 396–99. https://doi.org/10.3109/13880209409083022.
15
Rengasamy Ragupathi Raja Kannan, Rajasekaran Arumugam, and Perumal Anantharaman. “Antibacterial
Potential of Three Seagrasses against Human Pathogens.” Asian Pacific Journal of Tropical Medicine 3, no. 11
(2010): 890–93. https://doi.org/10.1016/s1995-7645(10)60214-3.
16
Rengasamy Ragupathi Raja Kannan, Rajasekaran Arumugam, and Perumal Anantharaman. “Chemometric
Studies of Multielemental Composition of Few Seagrasses from Gulf of Mannar, India.” Biological Trace Element
Research 143, no. 2 (March 2010): 1149–58. https://doi.org/10.1007/s12011-010-8911-y.
17
F. Folmer, M. Jaspars, M. Dicato, and M. Diederich. “Photosynthetic Marine Organisms as a Source of
Anticancer Compounds.” Phytochemistry Reviews 9, no. 4 (2010): 557–79. https://doi.org/10.1007/s11101-010-
9200-2.
18
Gordon Cragg, and David Newman. “Natural Product Drug Discovery in the Next Millennium.” Pharmaceutical
Biology 39, no. 1 (2001): 8–17. https://doi.org/10.1076/phbi.39.7.8.5868.
19
Rosane Oliveira. “Why Phytochemicals Are Important.” UC Davis Integrative Medicine, February 14, 2018.
https://ucdintegrativemedicine.com/2015/06/why-phytochemicals-are-important/#gs.3wknfj.
20
C. Nwabuisi, “Prophylactic Effect of Multi-Herbal Extract ‘Agbo-Iba on Malaria Induced in Mice.” East African
Medical Journal 79, no. 7 (January 2002). https://doi.org/10.4314/eamj.v79i7.8836.
present in phenols, flavonoids, terpenoids, glycosides, and saponins21.
Phytochemicals from seagrasses have been found to have significant functions such as the tannin cells being
specialized to produce phenolic compounds, which play defensive roles against microorganisms and
herbivores22,23,24. Chemical constituents of several seagrasses have been described including the antibiotic
flavone glycoside from T. testudium25, one sugar derivative from Ruppia maritime26, phenolic compounds
from P. oceanica27, diterpens from R. maritima28 and steroids and fatty acids from Zostera japonica23,29,30.
Apart from medicinal purposes, seagrasses also have other uses like the Enhalus acoroides which
is used as fishing nets in Moluccas and New Guinea. It is also used as mats, ropes and paper making. In the
Philippines and Indonesia, the seeds of E. acoroides are sometimes eaten31.
To date, little information exists about the phytochemical components of these species of
seagrasses. Hence, the goals of this study are to determine if flavonoids, tannins, saponins, phenols,
alkaloids, carbohydrates, proteins and amino acids, and coumarins are present in Enhalus acoroides,
Thalassia hemprichii, and Halodule uninervis.
Literature Review
Seagrasses in shallow sheltered regions of estuarine, brackish, and marine environments are of productive
and ecological importance. Because of this [M. D. Richmond, A Guide to the Seashores of Eastern Africa:
and the Western Indian Ocean Island (Zanzib́ ar, Tanzania: SIDA. Department for Research Cooperation,
SAREC, 1997)], talked about the status of a seagrass ecosystem in an ecologically sensitive wetland in
India. They discovered that in India, the seagrass habitat is mainly overlooked in terms of education, study
and management, although it is classified under ecologically delicate coastal regions. Although one of the
predominant macrophytic marine flora, seagrasses have surprisingly not been introduced in plant science
studies, even at the university level. In the recent past, the lack of awareness of the functions of seagrasses

21
P Tiwari, B Kumar, M Kaur, G Kaur, H Kaur. “Phytochemical screening and extraction: a review.” Int Pharm Sci
1, no. 1 (2011): 98-106.
22
Alice S. Tempel, “Tannin-Measuring Techniques.” Journal of Chemical Ecology 8, no. 10 (1982): 1289–98.
https://doi.org/10.1007/bf00987762.
23
J. Kuo and A.J. Mc Comb. Seagrass taxonomy structure and development. France: Elsevier Publication, 1989.
24
L. Cariello, and Laura Zanetti. “Distribution of Chicoric Acid during Leaf Development of Posidonia Oceanica.”
Botanica Marina 22, no. 6 (1979). https://doi.org/10.1515/botm.1979.22.6.359.
25
P. Jensen, K. Jenkins, D. Porter, W. Fenical, “Evidence that a new antibiotic flavone glycoside chemically defends
the sea grass Thalassia testudinum against zoosporic fungi.” Appl Environ Microbiol 64, no. 4 (1998): 1490–1496.
26
R. S. Aquino, “Occurrence of Sulfated Galactans in Marine Angiosperms: Evolutionary Implications.”
Glycobiology 15, no. 1 (2004): 11–20. https://doi.org/10.1093/glycob/cwh138.
27
James S. Todd, Richard C. Zimmerman, Phillip Crews, and Randall S. Alberte. “The Antifouling Activity of
Natural and Synthetic Phenol Acid Sulphate Esters.” Phytochemistry 34, no. 2 (1993): 401–4.
https://doi.org/10.1016/0031-9422(93)80017-m.
28
Marina Dellagreca, Antonio Fiorentino, Marina Isidori, Pietro Monaco, and Armando Zarrelli. “Antialgal Ent
Labdane Diterpenes from Ruppia Maritima.” Phytochemistry 55, no. 8 (2000): 909–13.
https://doi.org/10.1016/s0031-9422(00)00253-3.
29
Francis T. Gillan, Ronald W. Hogg, and Edward A. Drew. “The Sterol and Fatty Acid Compositions of Seven
Tropical Seagrasses from North Queensland, Australia.” Phytochemistry 23, no. 12 (1984): 2817–21.
https://doi.org/10.1016/0031-9422(84)83021-6.
30
Nina M. Sanina, Svetlana N. Goncharova, and Eduard Y. Kostetsky. “Fatty Acid Composition of Individual Polar
Lipid Classes from Marine Macrophytes.” Phytochemistry 65, no. 6 (2004): 721–30.
https://doi.org/10.1016/j.phytochem.2004.01.013.
31
“Enhalus Acoroides (PROSEA).” Enhalus acoroides (PROSEA) – PlantUse English. Accessed September 22,
2019. https://uses.plantnet-project.org/en/Enhalus_acoroides_(PROSEA).
at educational level and among the common people and coastal zone managers has caused enormous
damage to them. Seagrass habitat is under constant threat from various anthropogenic activities3.
On the other hand, phytochemicals have contributed a lot in the medicine industry. The study [P
Tiwari, B Kumar, M Kaur, G Kaur, H Kaur. “Phytochemical screening and extraction: a review.” Int Pharm
Sci 1, no. 1 (2011): 98-106] has contributed a lot in the phytochemical screening. It is about the methods
of phytochemical screening and extractions. They discovered that non-standardized methods of extraction
may be the cause of the degradation of the phytochemicals and may lead to the variations resulting to lack
of reproducibility21.
Meanwhile, the study [N. P. Bharathi, M. Jayalakshmi, P. Amudha, & V. Vanitha, “Phytochemical
screening and in vitro antioxidant activity of the seagrass Cymodocea serrulata.” Indian Journal of Geo
Marine Sciences 5, no.5 (2019): 1216-1221] talks about the phytochemical screening and in vitro
antioxidant activity of the seagrass Cymodocea serrulata. They determined the phytochemical constituents
and its antioxidant potential of Cymodocea serrulata. Their study brought out the medicinal value of C.
serrulata inspiring other researchers to study more of the uses of C. serrulata32.
Methodology
Collection of seagrasses. E. acoroides, T. hemprichii, and H. uninervis were collected in Brgy. Pagnamitan
in the month of September. Two hundred grams of each seagrass species was collected with the assistance
of a marine biologist from DENR-CENRO Guiuan sub-office for the identification. The collected seagrass
was placed in a cloth bag and brought to the laboratory. Samples of each specimen was sent to Visayas
State University for species verification.
Preparation of Seagrass. The seagrasses were washed with running tap water along with stroking the
leaves to remove the algae. This process was repeated twice for each species. Then it was rinsed with
distilled water. The seagrasses were air dried using a net. It took 7 days for the seagrasses to completely
dry.
Extraction of seagrasses. The leaves of every species were grinded using a blender. For the flavonoids,
alkaloids, carbohydrates, proteins and amino acids, and coumarins tests, the extraction of maceration
method was used21. Ten g of the grinded dried leaves of E. acoroides were mixed with a 100mL distilled
water in an Erlenmeyer flask33. The process was repeated for the T. hemprichii and H. uninervis. The flasks
were sealed with aluminum foil and stored in a cabinet to homogenize for 24 hours. After 24 hours, it was
filtered using Whatman (no.1) filter paper. The filtrate was placed in petri dishes and left to evaporate
through an improvised water bath in absence of a rotary evaporator at around 40°C until all the solvent was
totally removed21. Same process was done to the T. hemprichii and H. uninervis. For the tannins, saponins,
and phenols tests, 0.5g of the grinded dried leaves of E. acoroides was mixed with 20 ml distilled water
and heated until boiled. After boiling, it was filtered with Whatman no.1 filter paper 21. The same process
was done to the T. hemprichii and H. uninervis.

32
N. P. Bharathi, M. Jayalakshmi, P. Amudha, & V. Vanitha, “Phytochemical screening and in vitro antioxidant
activity of the seagrass Cymodocea serrulata.” Indian Journal of Geo Marine Sciences 5, no.5 (2019): 1216-1221.
33
M. Luay, M. Gonzaga, S. Po, and E. Arollado. “Determination of the Antiangiogenic Activity of Telecospium
telecospium Extract using in ovo Chorioallantoic Membrane Assay.” Acta Medica Philippina 52, no. 4 (2018): 366
Phytochemical Screening
Detection of Flavonoids. Alkaline Reagent Test was used. Crude extract was dissolved in 20mL of distilled
water. Three mL of the aqueous extract was placed in each of the 4 test tubes and labeled trial 0, 1, 2, and
3. Trial 1-3 were treated with 4 drops of 2% sodium hydroxide solution. Formation of intense yellow color,
which becomes colorless on addition of acetic acid, indicates the presence of flavonoids21.
Detection of Tannins. Ferric chloride Test was used. Three mL of the aqueous extract was placed in each
of the 4 test tubes and labeled trial 0, 1, 2, and 3. Trial 1-3 were treated with 4 drops of ferric chloride.
Formation of bluish black and brownish green coloration indicates the presence of tannins21.
Detection of Phenols. Ferric chloride Test was used. Three mL of the aqueous extract was placed in each
of the 4 test tubes and labeled trial 0, 1, 2, and 3. Trial 1-3 were treated with 4 drops of ferric chloride.
Formation of bluish black coloration indicates the presence of phenols21.
Detection of Saponins. Foam test was used. Three mL of the aqueous extract was placed in each of the 4
test tubes and labeled trial 0, 1, 2, and 3. 2mL of distilled water was added to trial 1-3. The mixture was
shaken vigorously. Formation of foam indicates the presence of saponins21.
Detection of Alkaloids. Wagner’s test was used. Extracts was dissolved in dilute Hydrochloric acid and
filtered. Three mL of the aqueous extract was placed in each of the 4 test tubes and labeled trial 0, 1, 2, and
3. Trial 1-3 were treated with 4 drops of Wagner’s reagent (Iodine in Potassium Iodide). Formation of
brown/reddish precipitate indicates the presence of alkaloids21.
Detection of Carbohydrates. Crude extract was mixed with 20mL distilled water. Three mL of the aqueous
extract was placed in each of the 4 test tubes and labeled trial 0,1,2, and 3. Trial 1-3 were treated with 4
drops of iodine solution. A dark blue or purple coloration indicates the presence of the carbohydrate34.
Detection of Proteins and Amino Acids. Xanthoproteic Test was used, crude extract was mixed with
20mL distilled water. Three mL of the aqueous extract was placed in each of the 4 test tubes and labeled
trial 0, 1, 2, and 3. Trial 1-3 were treated with 4 few drops of conc. Nitric acid. Formation of yellow color
indicates the presence of proteins21.
Detection of Coumarins. The method of Rizk, A.M., was used. Crude extract was mixed with 20mL
distilled water. Two mL of the aqueous extract was placed in each of the 4 test tubes and labeled trial 0, 1,
2, and 3. A 3 ml of 10% NaOH was added to 2 ml of aqueous extract. Formation of yellow color indicates
the presence of coumarins35.
Risk and Safety
The collection of seagrasses was performed in a shallow intertidal flat with minimal to none
swimming skill under the close supervision of adult. In handling the chemicals that was used in the
phytochemical screening, the researcher used a laboratory gown, mask, and gloves, while the preparation
of reagents will be performed in a fume hood with the assistance of a licensed chemist. For the disposal of

34
R. Yadav, Agarwala, M. “Phytochemical analysis of some medicinal plants.” Journal of Phytology 3, no. 12
(2011): 10-14.
35
N. Savithramma, M. Linga Rao, D. Suhrulatha, “Screening of Medicinal Plants for Secondary Metabolites.”
Middle East Journal of Scientific Research 8, no.3 (2011): 579-584.
the reagents and treated extracts, it was poured slowly in the sink along with running tap water to dilute the
chemicals.
Data Analysis
This study followed a completely randomized research design (CRD), for each set up received each
treatment having 4 trials, Trial 0,1, 2 and 3. Trial 0 being a comparative group was treated with distilled
water for comparison to the results of Trials 1, 2 and 3. Positive sign (+) was used to indicate the presence
of the phytochemical based on the color change as indicator of the tests was observed, while negative sign
(-) was used to indicate the absence of the phytochemical when no color change was observed on the extract.
Results and Discussion
Table 1. Test Results in Detecting Eight Phytochemicals from the Selected Seagrasses

Setups
Tests Trials
E. acoroides T. hemprichii H. uninervis
Trial 0 - - -
Trial 1 + + -
Flavonoids Test
Trial 2 + + -
Trial 3 + + -
Trial 0 - - -
Trial 1 + + +
Tannins Test
Trial 2 + + +
Trial 3 + + +
Trial 0 - - -
Trial 1 + - -
Saponins Test
Trial 2 + - -
Trial 3 + - -
Trial 0 - - -
Trial 1 + + +
Phenols Test
Trial 2 + + +
Trial 3 + + +
Trial 0 - - -
Trial 1 + + +
Alkaloids Test
Trial 2 + + +
Trial 3 + + +
Trial 0 - - -
Trial 1 - - -
Carbohydrates Test
Trial 2 - - -
Trial 3 - - -
Trial 0 - - -
Proteins and Trial 1 - - -
Amino Acids Test Trial 2 - - -
Trial 3 - - -
Trial 0 - - -
Trial 1 - - -
Coumarins Test
Trial 2 - - -
Trial 3 - - -
Results and Discussion
Table 1 shows the test result in detecting flavonoids, tannins, phenols, saponins, alkaloids, carbohydrates,
proteins and amino acids, and coumarins from E. acoroides, T. hemprichii, and H. uninervis. The three
seagrasses resulted positive on the tannins, phenols and alkaloids tests, E. acoroides and T. hemprichii on
the flavonoids test, and only E. acoroides saponins test for the color change indicator was observed.
Contrarily, the three seagrasses resulted negative on the carbohydrates, proteins and amino acids, and
coumarins tests, T. hemprichii and H. uninervis on the saponins, and only H. uninervis on the flavonoids
test for the expected color change was not observed.
Flavonoids are important antioxidants, and promote several effects. Aside from antioxidant activity,
flavonoids also provide anti-viral, anti-cancer, anti-inflammatory, and anti-allergic activities36. In the
flavonoids test, E. acoroides, and T. hemprichii, resulted positive for the color change indicator was
observed compared to trial 0.
Plants containing tannins have astringent, hemostatic, antiseptic and toning properties. It is also used for
small hemorrhages, sore mouth, bronchitis, burns, etc. Tannins also have antioxidant properties37. In the
tannins test, E. acoroides, T. hemprichii, and H. uninervis resulted positive for the color change indicator
was observed compared to trial 0.
Saponins have lots of health benefits. Some of these are cholesterol reduction, reduce cancer risk, immunity
booster, reduce bone loss, and antioxidants38. In the saponins test, only E. acoroides resulted positive for
the color change indicator was observed compared to trial 0.
The health benefits of phenols are antioxidants, cancer prevention, sore throat spray, vaccine preservative,
etc39. In the phenols test, E. acoroides, T. hemprichii, and H. uninervis resulted positive for the color change
indicator was observed compared to trial 0.
Some of the uses of alkaloids are as medicines, and as pesticide or insect repellents. Uses of alkaloids is
mainly used in health care. They act as lifesaving drugs in some diseases like heart-failure, cancer, blood
pressure etc40. In the alkaloids test, E. acoroides, T. hemprichii, and H. uninervis resulted positive for the
color change indicator was observed compared to trial 0.
Conclusion
Based on the test results, E. acoroides contain flavonoids, tannins, saponins, phenols, and alkaloids.
While T. hemprichii flavonoids, tannins, phenols, and alkaloids and H. uninervis flavonoids, tannins,
phenols, and alkaloids.

36
Hasen Muhaisen, “Introduction and Interpretation of Flvonoids.” Advanced Science, Engineering and Medicine 6,
no. 12 (2014): 1235-1250. https://doi.org/10.1166/asem.2014.1630.
37
“Tannins.” Pharmacognosy. Accessed September 22, 2019. https://www.medicinalplants-
pharmacognosy.com/pharmacognosy-s-topic/tannins/.
38
Saponins. Accessed September 22, 2019. https://www.phytochemicals.info/phytochemicals/saponins.php.
39
Tim Jewell. “What Is Phenol? Medicinal Uses, Health Benefits, and Risks.” Healthline. Healthline Media, January
25, 2019. https://www.healthline.com/health/what-is-phenol#benefits.
40
“5 Uses of Alkaloids in Medicine, Agriculture & Research.” Study Read, November 22, 2018.
https://www.studyread.com/uses-of-alkaloids/.
 Tannins & Phenols Test:
T. hemprichii E. acoroides H. uninervis

Alkaloids Test
T. hemprichii E. acoroides H. uninervis

Carbohydrates Test
T. hemprichii E. acoroides H. uninervis

Coumarins Test
T. hemprichii
 E. acoroides

H. uninervis

Flavonoids Test
T. hemprichii

E. acoroides

H. uninervis

Proteins and Amino acids

T. hemprichii
E. acoroides

H. uninervis