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ISSN: 2329-9002

Research Article OMICS International

Preliminary Phytochemical Screening and Toxicity Test of Leaf and Root

Parts of the Snake Plant (Sansevieria trifasciata)
Julie S. Berame*, Sheena Mae E. Cuenca, Diana Rose P. Cabilin and Marycris L. Manaban
Department of Biology, College of Education, Caraga State University, Ampayon, Butuan City, 8600 Philippines
*Corresponding Author: Julie S. Berame, Department of Biology, College of Education, Caraga State University, Ampayon, Butuan City, 8600 Philippines, Tel: +63
9293950188; E-mail: [email protected]
Receiving date: Sep 26, 2017; Acceptance date: Oct 13, 2017; Publication date: Oct 18, 2017
Copyright: © 2017 Berame JS, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted
use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Objective: To examine the toxicity level of Sansevieria trifasciata (Asparagaceae) leaves and roots extracts.

Methods: The antimicrobial activity was tested against alkaloid, saponins, tannins, anthraquinones screening by
two different methods, the brine shrimp toxicity test and ten-fold serial dilution of the powdered plant material in
artificial seawater.

Results: The results showed the presence of cytotoxic principles making it effective therapeutic remedy for
treating various infections as it possesses effective of its zone of inhibition (lowest concentration of 35.22 μg/ml to
the highest concentration of 44.49 μg/ml) using brine shrimp bioassy of Sansevieria trifasciata roots and leaves
extracts.

Conclusion: Thus, the toxic potential of the plant extracts yields greater than the recommended LC50 value and
reveals positive linear relationship between the concentrations of the extract to the mortality rate using nauplii. The
more concentrated the treatment the higher mortality had resulted.

Keywords: Phytochemical screening; Brine shrimp; Lethality test; This research would help in the discovery of a new form of
Secondary metabolites; Snake plant medication for the upcoming severe diseases of microbial infection
such as the leaves and roots of Sansevieria trifasciata are used in
Introduction traditional medicine for the treatment of asthma, abdominal pains,
colic, diarrhea, haemorrhoids, hypertension, menorrhagia, piles, sexual
Snake Plant is one of the most recommended plants for improving weakness, wounds of the foot, cough, leprosy, rheumatism, glandular
air quality. The optimal place to keep this is in your bedroom, because enlargement, nutritional deficiencies and treatment of snake bite [4].
it converts CO2 into oxygen at night it also removes contaminants The active ingredient from the root is more natural, fewer chemicals so
from air. The NASA conducts a study to the uses of snake plant it consists less toxicity, side effects and safe compared to synthetic
according to their results they found out that the snake plant absorbs drugs [5]. The new discovery of this product will create awareness and
toxins, such as nitrogen oxides and formaldahyde. Sansevieria revolute advantage of the Sansevieria trifasciata root usage [6,7]. This
trifasciata is able to absorb 107 types of toxins, including air pollution, medication well tolerated by patients, with fewer unintended
cigarette smoke (nicotine), so it would make a great refresher. It also consequences than pharmaceutical drugs because it less toxic and side
contains phytochemicals such as flavonoids, saponins, and glycosides effects [8]. Thus, this study will be an important way of labelling the
which reduce the number of bacteria [1]. In China, decoction used for snake plant abundantly found in as ornamental plant in Caraga State
detoxification, as anti-inflammatory, and for treatment of sores and University. Its bioactive ingredients were identified and its toxicity level
snake bites. It also used to cure boils, cough, bronchitis, traumatic for the effective future formulation of the plant as medicinal plant.
injuries [2].
The search for bioactive compounds for pharmaceutical and Materials and Methods
industrial applications is still being actively done. Philippine plant
species that have been used for indigenous medicinal purpose serves as Plant materials
a reservoir of these compounds. At present, herbal medicines and food
supplements are undeniably rampant as an alternative to synthetic The Fresh leaves and roots of Snake plant were collected from the
drugs. Media advertisements as well as beauty cosmetics brochures and Engineering Department of Caraga State University along its side
catalogues are flooded with varieties of botanicals and herbals. This is pavement and across the Horrizon Bridge on its paved aisle along the
caused by widely accepted through that these products have no side its pavement.
effects, can easily absorbed by the body and can cure almost all of the The leaves of Sansevieria liberica were collected in July 2017 from a
common ailments; that is in comparison to synthetic drugs [3]. single population of the plant from front of the Engineering
However, labels and packages of these sold herbal medicines states that Department of Caraga State University, Ampayon, Butuan City. The
it has no approved therapeutic claim. collected plant was authenticated with the biology department.

J Phylogenetics Evol Biol, an open access journal Volume 5 • Issue 3 • 1000187

ISSN: 2329-9002
Citation: Berame JS, Cuenca SME, Cabilin DRP, Manaban ML (2017) Preliminary Phytochemical Screening and Toxicity Test of Leaf and Root
Parts of the Snake Plant (Sansevieria trifasciata). J Phylogenetics Evol Biol 5: 187. doi:10.4172/2329-9002.1000187

Page 2 of 7

Preparation extract shaken. The formation of a pink, red or violet colour on the
ammoniacal phase [10].
The collected plant parts were separated as leaf and roots and were
cleaned with deionized water and dried under shade for two weeks at
room temperature. Dried leaves and stem were grounded and filtered
Brine shrimp bioassay
using 0.3 mm mesh. The plant powder was stored in air tight container The brine shrimp assay is a rapid, reliable and inexpensive general
and maintained at 4°C until use separately. bioassay toxicity test for active plant extracts. The procedure allows the
determination of the LC50 values in parts per million (ppm) of active
Qualitative phytochemical screening constituents in the brine medium. This bioassay has been used in the
analysis of natural products, pesticides residues and other chemical
Phytochemical analysis of each extract has been carried out pollutants in marine environment [11].
according to standard protocols [9].
Hatching of brine shrimp
Screening for alkaloid
Brine Shrimp egg, (Artemia salina) obtained from the local pet shop
A 0.5 g of the extract was stirred in 5 ml of 1% HCl on a steam bath was hatched in a shallow rectangular plastic dish filled with 1 liter of
and filtered while hot. Distilled water was added to the residue and 1 artificial sea water, which was prepared with 35 grams commercial
ml of the filtrate was treated with a few drops of Wagner’s reagent. A rock salt mixture and 1 mL distilled water. Approximately 1 gram of
reddish brown precipitate indicates the presence of alkaloids. eggs was sprinkled in the compartment. An aquarium aerator was used
Screening for Flavonoids 2 ml of dilute sodium hydroxide was added for Artemia cyst suspension to provide an ample supply of oxygen in
to 2 ml of the extract. The appearance of a yellow colour indicates the the medium. A perforated plastic cover was then put over the dish to
presence of flavonoids. avoid predatory insects on cysts or nauplii. The container was also well
lighted with lamp to maintain the water temperature. After 3 days the
Screening for saponins hatched cysts have produced fast swimming nauplii which were used
A 1 ml of distilled water was added to 1 ml of the extract and shaken in this bioassay. A dropper and a micropipette were used to collect
vigorously. A stable persistent froth indicated the presence of saponins. nauplii from the suspension.
Screening for Phenols Equal volumes (1 ml) of extract and Iron (III)
chloride were mixed. A deep bluish green solution gave an indication Ten-fold serial dilution of the powdered plant material in
of the presence of phenols. artificial seawater
Three test tubes containing 0.1 g or 100 mg of each powdered snake
Screening for tannins leaves and roots, 10 mL of artificial water was introduced and the test
A portion of the extract was dissolved in water, after which the tube was corked and manually shaken by inverting 5 times. The
solution was clarified by filtration. 10% ferric chloride solution was concentration of the snake plant extract in this tube is 10 mg/ml or
then added to the resulting filtrate. The appearance of a bluish black 10,000 ứg/mL. Using a permanent marker, this tube was labeled # 1.
colour indicates the presence of tannins. Five other tubes were labeled 2, 3, 4, 5 & 6. The test tubes were placed
in a rack side by side according to their number. To tubes # 2,3,4,5
Screening for anthraquinones artificial seawater of 9 mL volume was added using 10 mL glass pipette
and tube # 6 was 10 mL artificial seawater and serves as control set-up.
A 0.5 g of the extract was shaken with 10 ml of benzene and filtered.
10% of ammonia solution was added to filtrate and the mixture was

Tube # #1 #2 #3 #4 #5

Volume of ASW (Diluent) 10 mL 9 mL 9 mL 9 mL 9 mL

Volume of Original Plant Extract (10 mg/mL) Serially Diluted ____ 1.0 mL 1.0 mL 1.0 mL 1.0 mL

Total volume in tubes # 1 to # 5 10 mL 10 mL 10 mL 10 mL 10 mL

Dilution Series 10-fold 10-fold 10-fold 10-fold 10-fold

Dilution Level of Originated Extract 1 1:10 1:100 1:1,000 1:10,000

Concentration of Plant extract in mg/mL from tubes # 1 to # 5 10 1.0 0.1 0.01 0.001

Concentration of plant extract in ứg/mL or ppm from tube # 1 to # 5 10, 000 1, 000 100 10 1

Table 1: Ten-fold dilution series of the Three dried powdered specie of Sansevieria trifasciata in ASW of various concentrations ranging from 10,
000 ứg/mL to 1 ứg/mL. Note: # denotes Number.

The plant powder in tube # 1(10 mg/mL) was allowed to settle down bottom of tube # 1, 1 mL of the supernatant was gently withdrawn
for 1 hour. This should be produced a residue at the bottom and a clear using a 1 mL micropipettor and transferred to tube # 2. Tube # 2 was
supernatant liquid above it. After the plant powdered has settle to the corked and inverted 3 times to mix the content. Tube # 2 was uncorked

J Phylogenetics Evol Biol, an open access journal Volume 5 • Issue 3 • 1000187

ISSN: 2329-9002
Citation: Berame JS, Cuenca SME, Cabilin DRP, Manaban ML (2017) Preliminary Phytochemical Screening and Toxicity Test of Leaf and Root
Parts of the Snake Plant (Sansevieria trifasciata). J Phylogenetics Evol Biol 5: 187. doi:10.4172/2329-9002.1000187

Page 3 of 7

and return to its slot in the rack between # 1 and # 3. Using again the � ∑ �� − ∑ � ∑ �
micropipettor, 1 mL was removed from tube # 2 and transferred to �=
� �2 − (�)2
∑ ∑
tube # 3 to tube # 5. This process performed is a 10-fold serial dilution
indicating that in each tube as one move from tube # 1 to # 5, the The Coefficient from the equation of a line above is solved as a=y-bx
concentration of the plant extract is being reduced by 10 times of the Where y is the mean of y (percent mortality) and x is the mean of x
tube previous to it shows in Table 1. (log dose).
Assuming that y=50% and substituting it to the regression equation
Bioassay above and then x was solved. And with a scientific calculator, antilog
In each snake plant extract, 18 vials contained with 5 mL ASW were values achieved were converted it back to its dose in ppm to determine
prepared for the application. Approximately 10 shrimps were collected the LC50 of each snake plant extract in ppm. The resulting values were
using a wide-mouth plastic medicine dropper. The shrimps were added the concentration of the snake plant species that will kill 50% of the
to each vial. A magnifying lens was used for checking the viability and Artemia salina nauplii at 24 hours. Then correlation coefficient of the
well-being of the nauplii. The plastic container was closed with a results on the data obtained was solved using the equation
plastic cellophane cover and kept under white light during the 24
� ∑ �� − ∑ � ∑ �
hours period. The numbers of viable nauplii (number of surviving) �=
were counted in each vial after 24 hours. A pipette sucked into the vials �∑ �2 − ∑ (�)2 − � ∑ �2 − ∑ (�)2
was used to count the surviving shrimps macroscopically, held against
a well-lighted background. Furthermore, the toxicity rating of sample as also analysed
according to the (%) mortality group of the test organism which is
show in Table 2 [12].
Statistical analysis to determine the extracts LC50
To determine the regression of equation for the data, the Coefficient
b in the equation of a line y=a+bx is computed as

Mortality Toxicity

80-90 Highly toxic

50-79 Moderately toxic

30-49 Slightly toxic

0-29 Non-toxic

Table 2: The toxicity rating of extract based on percent (%) mortality

Results
Weigh 0.1 gram of the sample then dry it in oven at 50oC for about 4
to 6 hours until constant weight is obtained. Powderized the sample for Phytochemical screening of ethanolic extract
the application in the tenfold dilution and for brine shrimp bio assay
The Table 3 and Table 4 below showed the result of the presence of
using the nauplii larvae.
the bioactive compounds found in both the leaves and root parts of the
snake plant. It can be seen that both the leaves and roots were highly
Bioassay procedure active with alkaloids, tannins and anthraquinones and two bioactive
compounds were absent in the leaves and roots of plants were
The procedure described in the literature [13] (Meyer et al. and flavonoids and saponins.
McLaghin et al.) was adopted for this study. Samples of five different
concentrations 1, 10, 100, 1000, 10000 µg/ml were prepared according Ten-fold serial dilution of the powdered plant material in
to the direction given in the literature. Brine shrimp (Artemia salina
artificial seawater
Leach) nauplii were hatched in a specific plastic tank (Figure 1). Fifteen
shrimps were transferred to each sample vial and then sea water was Three test tubes containing 0.1 g or 100 mg of each powdered snake
added to make the volume of 2 ml. The vials were kept for 24 hours, leaves and roots, 10 mL of artificial water was introduced and the test
thereafter the active nauplii were counted with the aid of 3x tube was corked and manually shaken by inverting 5 times. The
magnifying glass and the percent death at each dose and control was concentration of the snake plant extract in this tube is 10 mg/ml or
determined. The data obtained were processed in Probit Statistical 10,000 ứg/mL. Using a permanent marker, this tube was labeled # 1.
Analysis by means of regression to estimate LC50 values for the Five other tubes were labelled 2, 3, 4, 5 & 6. The test tubes were placed
significance of death, survival and the mortality of nauplii larvae. in a rack side by side according to their number. To tubes # 2,3,4,5
artificial seawater of 9 mL volume was added using 10 mL glass pipette
and tube # 6 was 10 mL artificial seawater and serves as control set-up.

J Phylogenetics Evol Biol, an open access journal Volume 5 • Issue 3 • 1000187

ISSN: 2329-9002
Citation: Berame JS, Cuenca SME, Cabilin DRP, Manaban ML (2017) Preliminary Phytochemical Screening and Toxicity Test of Leaf and Root
Parts of the Snake Plant (Sansevieria trifasciata). J Phylogenetics Evol Biol 5: 187. doi:10.4172/2329-9002.1000187

Page 4 of 7

Test for Test for Antraquinones Test for

Test for Alkaloids Test for Tannins
Flavonoids Saponins

Plant Sample Dragen- Ferric Chloride Smith & Metcalf Borntrager’s Test and Froth Test
Mayers Reagent dorffs Gelatin Test Test Method Modefied Borntrager’s Test
Reagent

Snake Plant Leaves Brownish brown _ + _

++ ++ ++
Extract +

Snake Plant Roots

+++ +++ ++ ++ _ ++ _
Extract

Table 3: Phytochemical Screening of the Ethanolic Extract of Snake Plant (Sansevieria Trifasciata). +++ (Highly present); ++ (Moderately
present); + (present) and - ( absent)

Ten-fold dilution test

Tube # microgram/ml #1 #2 #3 #4 #5

Volume of ASW (Diluent) 10 mL 9 mL 9 mL 9 mL 9 mL

Volume of Original Plant Extract (10 mg/mL) Serially Diluted ____ 1.0 mL 1.0 mL 1.0 mL 1.0 mL

Total volume in tubes # 1 to # 5 10 mL 10 mL 10 mL 10 mL 10 mL

Dilution Series 10-fold 10-fold 10-fold 10-fold 10-fold

Dilution Level of Originated Extract 1 1:10 1:100 1:1,000 1:10,000

Concentration of Plant extract in mg/mL from tubes # 1 to # 5 10 1.0 0.1 0.01 0.001

Concentration of plant extract in ứg/mL or ppm from tube # 1 to # 5 10, 000 1, 000 100 10 1

Table 4: Ten-fold dilution series of the Three dried powdered species of Sansevieria trifasciata in ASW of various concentrations ranging from 10,
000 ứg/mL to 1 ứg/mL.

The plant powder in tube # 1(10 mg/mL) was allowed to settle down and return to its slot in the rack between # 1 and # 3. Using again the
for 1 hour. This should be produced a residue at the bottom and a clear micropipettor, 1 mL was removed from tube # 2 and transferred to
supernatant liquid above it. After the plant powdered has settle to the tube # 3 to tube # 5. This process performed is a 10-fold serial dilution
bottom of tube # 1, 1 mL of the supernatant was gently withdrawn indicating that in each tube as one move from tube # 1 to # 5, the
using a 1 mL micropipettor and transferred to tube # 2. Tube # 2 was concentration of the plant extract is being reduced by 10 times of the
corked and inverted 3 times to mix the content. Tube # 2 was uncorked tube previous to it (Table 5).

Well # Conc. of Log 10 Initial nos. of Shrimps surviving Dead shrimps % Mortality % Mortality
shrimps (corrected for
Snake Plant Leaves Conc. Of = dead shrimps
control)
÷initial no. of
Extract Snake plant
shrimps x 100
µg/ml Leaves
Extract

1 10,000 4 45 7 38 84 84-11=73

2 1,000 3 45 12 33 73 73-11=62

3 100 2 45 15 30 67 67-11=56

4 10 1 45 20 25 56 56-11=45

5 1 0 45 26 19 42 42-11=31

Control 0 (control) ------- 45 40 5 11 --------

J Phylogenetics Evol Biol, an open access journal Volume 5 • Issue 3 • 1000187

ISSN: 2329-9002
Citation: Berame JS, Cuenca SME, Cabilin DRP, Manaban ML (2017) Preliminary Phytochemical Screening and Toxicity Test of Leaf and Root
Parts of the Snake Plant (Sansevieria trifasciata). J Phylogenetics Evol Biol 5: 187. doi:10.4172/2329-9002.1000187

Page 5 of 7

Lethal Concentration @50 (LC50) = 44.49 µg/ml (Highly Toxic)

Table 5: Brine shrimp Bioassay for Snake Plant Leaves Extract Legend: 0-50 ppm (Highly Toxic); 51-100 ppm (Moderately Toxic); 101-200 ppm
(Slightly Toxic); and 201 and above Not Toxic.

Brine shrimp bioassay test antibacterial agent due to the high toxicity level as shown in the Table 4
above. This only means that the root extract part is more effective
Table 5 showed the bioassay using brine shrimp in the leaves extract herbal medicine compared with leaves extract since the number of
of snake plant. It showed a high level of cytotoxicity level of 44.49 surviving shrimps are more than the root extract’s number of surviving
µg/ml. This means that leaves extract of snake plant is an effective shrimps as shown in Table 6 below.
medicinal plant and can be good medicinal plant especially as

Well # Conc. of Log 10 Initial nos. of Shrimps surviving Dead shrimps % Mortality % Mortality
shrimps (corrected for
Snake Plant Roots Conc. of = dead shrimps
control)
÷initial no. of
Extract Snake Plant
shrimps x 100
Roots
µg/ml
Extract

1 10,000 4 45 5 40 89 89-11=78

2 1,000 3 45 10 35 78 78-11=67

3 100 2 45 12 33 73 73-11=62

4 10 1 45 18 27 60 60-11=49

5 1 0 45 20 25 56 56-11=45

Control 0 (control) ------- 45 40 5 11 --------

Lethal Concentration @50 (LC50) = 35.22 µg/ml (Highly Toxic)

Table 6: Brine shrimp Bioassay for Snake Plant Roots Extract Legend: 0-50 ppm (Highly Toxic); 51-100 ppm (Moderately Toxic); 101-200 ppm
(Slightly Toxic); and 201 and above Not Toxic.

Mortality rate of Nauplii Larvae that it only proved that it can be less effective when its extracts are
applied to these organisms and these events will be best observed or
This table showed that from the values of snake plant for its LC50 shown in Figure 1 below as the graph interpretation of comparing the
for roots of 35.22 µg/ml and LC50 of 44.49 µg/ml the brine shrimp Tables 4 and 5 respectively.
bioassay, the root part is less effective when applied as an herbal
medicine compared due to the number of less surviving shrimps and

Figure 1: The Comparison between the Toxicity Levels of Snake Plant

This figure showed the comparative status of explaining the toxicity

level of both extracts (plants and leaves) and will explain why leaves

J Phylogenetics Evol Biol, an open access journal Volume 5 • Issue 3 • 1000187

ISSN: 2329-9002
Citation: Berame JS, Cuenca SME, Cabilin DRP, Manaban ML (2017) Preliminary Phytochemical Screening and Toxicity Test of Leaf and Root
Parts of the Snake Plant (Sansevieria trifasciata). J Phylogenetics Evol Biol 5: 187. doi:10.4172/2329-9002.1000187

Page 6 of 7

extract is more effective as shown in Table 4 and why the roots extract as it possesses effective zone of inhibition. The toxicity of the extract
is less effective compared to the leaves extracts as shown in Table 5. when diluted with ethanol both the roots and leaves of the ST, its
This is the graph interpretation of the Tables 4 and 5 above. concentration showed that snake plant species can kill 50% of the
Artemia salina nauplii at 24 hours, as a result from its test thru
Discussion Brineshrimp Bioassay of its roots and leaves. As to the degree of
lethality aspect of the study, the extractives were found to be directly
The screening of the plant material revealed the presence of the proportional to the concentration of the extractives ranging from the
alkaloids, tannins, and antrhaquinones were summarized in Table 3. lowest concentration (35.22 μg/ml) to the highest concentration (44.49
Ethanol extract yielded almost all above mentioned phytochemicals. μg/ml). This concentration is an dependent increment in percent
Since our plant extract have revealed the presence of alkaloids, we can mortality of Brine Shrimp nauplii produced by the Snake plant and
say alkaloids also have shown the antibacterial activity [14]. which indicates the presence of cytotoxic principles in these extractives
The evaluated antibacterial activity of alkaloid of Datura metel Linn hence, making it as an effective therapeutic remedy for gastric ulcer
leaves against Staphylococcus aureus, Pseudomonas aeruginosa, and will lend the credence to the folkloric use of these plants in
Proteus mirabis, Solmonella typhi, Bacillus subtilis and Klebsiella treating microbial infection.
pneumonia but could not inhibit Escherichia coli [15]. Flavonoids are
ubiquitous in photosynthesing cells and are commonly found in fruit, Conflict of Interest Statement
vegetables, nuts, seeds, stems, flowers, tea, wine, propolis and honey.
We declare that we have no conflict of interest.
For centuries, preparations containing these compounds as the
principal physiologically active constituents have been used to treat
human diseases [16]. Acknowledgements
Increasingly, this class of natural products is becoming the subject of The researcher wish to express his recognition to the Department of
anti-infective research, and many groups have isolated and identified Biology and Chemistry Laboratory of Caraga State University for
the structures of flavonoids possessing antifungal, antiviral and allowing the researcher to use the laboratory apparatus.
antibacterial activity. The n-butanol purified saponin extract of
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ISSN: 2329-9002
Citation: Berame JS, Cuenca SME, Cabilin DRP, Manaban ML (2017) Preliminary Phytochemical Screening and Toxicity Test of Leaf and Root
Parts of the Snake Plant (Sansevieria trifasciata). J Phylogenetics Evol Biol 5: 187. doi:10.4172/2329-9002.1000187

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