Phytochemical Analysis, Total Phenolic Content, Antioxidant and Antidiabetic Activity of Sansevieria cylindrica Leaves Extract

Abstract

Traditionally, Genus Sansevieria has been used for the treatment of various ailments in African countries since ancient time. In South Africa and tropical American countries, a crude drug obtained from Sansevieria trifasciata species is sold out in the local market to cure the snakebite and inflammatory conditions. In this study, chemical and pharmacological studies of Sansevieria cylindrica have been carried out. Phytochemical analysis of extracts of Sansevieria cylindrica leaves showed the presence of steroids, flavonoids, saponins, tannins, and phenolic acids. Methanol fraction was found to show maximum phenolic content. Ethanol extract and its methanol fraction exhibited significant antioxidant and antidiabetic activities. The ethanol extract inhibited 80.5%, whereas methanol fraction showed 83.6% inhibition of DPPH free radicals at 100 μg/mL concentration respectively. In addition, methanol fraction exhibited 57.9% inhibition of glucose-6-phosphatase enzyme at 100 μM concentration. Our study confirmed the traditional uses of Sansevieria cylindrica plants for the treatment of various diseases.

Full text

2455-0299 / JACS Directory©2017. All Rights Reserved
Cite this Article as: Tanveer Ahamad, Devendra Singh Negi, Mohammad Faheem Khan, Phytochemical analysis, total phenolic content, antioxidant and antidiabetic activity of Sansevieria cylindrica leaves
extract, J. Nat. Prod. Resour. 3(2) (2017) 134-136.
J. Nat. Prod. Resour. - Volume 3 Issue 2 (2017) 134–136
Share Your Innovations through JACS Directory
Journal of Natural Products and Resources
Visit Journal at http://www.jacsdirectory.com/jnpr
Phytochemical Analysis, Total Phenolic Content, Antioxidant and Antidiabetic Activity of
Sansevieria cylindrica Leaves Extract
Tanveer Ahamad1, Devendra Singh Negi2, Mohammad Faheem Khan1,*
1Department of Biotechnology, Era’s Lucknow Medical College & Hospital, Sarfarazganj, Lucknow – 226 003, UP, India.
2Department of Chemistry, HNB Garhwal University, Srinagar – 246 174, UK, India.
A R T I C L E D E T A I L S
A B S T R A C T
Article history:
Received 06 August 2017
Accepted 28 August 2017
Available online 01 September 2017
Traditionally, Genus Sansevieria has been used for the treatment of various ailments in African countries
since ancient time. In South Africa and tropical American countries, a crude drug obtained from
Sansevieria trifasciata species is sold out in the local market to cure the snakebite and inflammatory
conditions. In this study, chemical and pharmacological studies of Sansevieria cylindrica have been
carried out. Phytochemical analysis of extracts of Sansevieria cylindrica leaves showed the presence of
steroids, flavonoids, saponins, tannins, and phenolic acids. Methanol fraction was found to show
maximum phenolic content. Ethanol extract and its methanol fraction exhibited significant antioxidant
and antidiabetic activities. The ethanol extract inhibited 80.5%, whereas methanol fraction showed
83.6% inhibition of DPPH free radicals at 100 μg/mL concentration respectively. In addition, methanol
fraction exhibited 57.9% inhibition of glucose-6-phosphatase enzyme at 100 µM concentration. Our
study confirmed the traditional uses of Sansevieria cylindrica plants for the treatment of various
diseases.
Keywords:
Sansevieria cylindrical
Phytochemical Analysis
Total Phenolic Content
Antioxidant Activity
Antidiabetic Activity
1. Introduction
Medicinal plants have been identified and used as significant herbal
medicine all over world from prehistoric times for the treatment of many
illness conditions [1]. These medicines are prepared in the form of crude
drug or in pure form from seeds, berries, roots, leaves, bark, or flower
parts of various plants. The therapeutic activity of a plant is due to the
presence of complex chemical constituents in different parts, providing
certain therapeutic effects [2]. World Health Organization (WHO)
estimated that 80% of people worldwide rely on herbal medicines with
increasing interest because of public dissatisfaction with the cost of
prescribe medications, various side effects of synthetic medicines, non-
toxic nature, more affordable with lower cost and allows greater public
access to health information [3]. Plants are rich sources of different types
of secondary metabolites which are generally termed as compounds or
chemical constituents. These compounds are not only used directly as
therapeutic agents, but also as starting materials for the synthesis of drugs
or as models for pharmacologically active compounds [4]. Many of these
compounds have pharmacological activities and used in the treatment of
chronic and acute conditions and various ailments such as cardiovascular
disease, prostate problems, depression, inflammation, to boost the
immune system and antioxidant properties. Antioxidant activity is shown
by phenols or their oxygen-substituted derivatives such as tannins,
flavonoids, phenolic acid as well as compounds having hydroxyl
functionality [5]. Based on the traditional aspects of herbal medicine and
Sansevieria cylindrical (SC), this study was conducted to evaluate the
antioxidant as well as antidiabetic activity of leaf extracts of this plant.
SC belongs to the family Asparagaceae, commonly referred to as Spear
Sansevieria. It is a succulent and an evergreen perennial plant, native to
the subtropical regions of the African continent and cultivated in Egypt for
ornamental purposes [6]. It is also found in some part of India as an
ornamental plant. SC including other species has different chemical
constituents such as dicarboxylic acids, phenols, steroidal saponins,
saponins, homoisoflavanone, coumarins and ester of fatty acids [7].
Sansevieria species were investigated for many pharmacological activities,
such as antimicrobial, antioxidant, antitumor, and antidiabetic activities
and inhibition of the capillary permeability activity [8]. However, a survey
of the literature showed that no antioxidant and antidiabetic activities of
leaves of SC have been carried out. As part of our research work on the
phytochemical investigation of medicinal plants, we have reported
antioxidant and antidiabetic activities of extracts and their fractions of
leaves of SC.
2. Experimental Methods
2.1 Chemical and Instrumentation
All solvents (ethanol, methanol, dichloromethane hydrochloric acid,
sulfuric acid, chloroform, ammonia, glacial acetic acid, sodium hydroxide)
were purchased from SD fine chemical limited, Mumbai, India and were
used without further purification. All chemicals were of analytical grade.
1,1-Diphenyl-2-picrylhydrazyl (DPPH), Folin-Ciocalteu reagent, gallic
acid, ascorbic acid, anhydrous sodium carbonate (Na2CO3), Dragendorff’s
reagent, mercuric chloride, potassium iodide, iodine were purchased from
Sigma-Aldrich, Mumbai, India. Solvents were recovered by using a water
bath (Perfit India) and Buchi Rotavapor (R-300). Absorbance was
measured with the help of UV-VIS spectrophotometer (Systronic, model
059).
2.2 Plant Material
SC leaves were collected from the botanical garden, Department of
Botany, University of Lucknow, Lucknow, UP, India in January 2017. Plant
material was kindly confirmed and authenticated Dr. Alka Kumari,
Department of Botany, University of Lucknow, Lucknow-226007, UP, India
where one voucher specimen was deposited. The collected samples were
air-dried, powdered and kept in tightly-closed container for further
experiment.
2.3 Extraction and Fractionation
SC leaves (1.0 kg) were chopped into small pieces and air-dried at room
temperature over several days until dryness. The dried pieces were
powdered, weighed and percolated with 95% ethanol. It was repeated five
times with an interval of 3 days. After percolation, filtration was carried
out using Whatman filter paper. The combined filtrate of the alcoholic
extracts was concentrated using Buchi Rotavapor (Interface R-300) with
the speed set at 150 RPM and temperature at 45 °C. The concentrated
*Corresponding Author
Email Address: faheemkhan35@gmail.com (Mohammad Faheem Khan)
ISSN: 2455-0299

135
Tanveer Ahamad et al. / Journal of Natural Products and Resources 3(2) (2017) 134–136
Cite this Article as: Tanveer Ahamad, Devendra Singh Negi, Mohammad Faheem Khan, Phytochemical analysis, total phenolic content, antioxidant and an tidiabetic activity of Sansevieria cylindrica leaves
extract, J. Nat. Prod. Resour. 3(2) (2017) 134-136.
extract was removed from the round bottom flask with ethanol and
poured into weighed beakers. The alcoholic solvent was allowed to
evaporate up to dryness. The dried solid extract was collected, weighed
and used for fractionation. The ethanol extract (50 g) was triturated with
hexane (5.1 g) and the hexane insoluble portion was dissolved in water,
which was successively extracted with chloroform (500 ml x 3) and
methanol (500 mL x 3) which yielded fractions of chloroform (12.7 g),
methanol (20.5 g) and water (8.2 g).
2.4 Phytochemical Screening
Phytochemical screening was carried out for all the extracts and
fractions according to the method described by Trease and Evans [9], with
slight modifications. The screening was performed for steroids, flavonoids,
saponins, tannins, and phenolic acids. The color intensity or the precipitate
formation was used as analytical tests. The qualitative results are
expressed as (+) for the presence and (−) for the absence of
phytochemicals.
2.5 Total Phenolic Content
Total soluble phenolics of the extracts were determined with Folin-Cio
calteau’s reagent with the help of UV-VIS spectrophotometer [10]. Gallic
acid was used as a standard. One mg of Gallic acid was dissolved in 10 mL
of methanol (100 μg/mL) to prepare a stock solution and then further
diluted to 8, 4, 2 and 1 μg/mL. One mL aliquot of each dilution was taken
and diluted with 10 mL of distilled water. Then 3 mL Folin-Ciocalteu
reagent was added and allowed to incubate at room temperature for 5 min,
2 mL of 20% (w/w) Na2CO3 was added in each sample and left to stand for
30 min at room temperature. Absorbance was measured at 765 nm using
UV-VIS spectrophotometer against blank (distilled water). Results were
expressed as milligrams of gallic acid equivalent (GAE) per 100 g of the dry
sample. The coefficient of determination was r = 0.9968.
2.6 DPPH Assay
DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity was
evaluated based on the method described previously [11]. It was
measured a decrease in absorbance at 517 NM of a solution of colored
DPPH in methanol brought about by the sample. Ascorbic acid was used as
a reference compound. Briefly, 5 mL of methanol, DPPH (0.1 mM) was
added to 1 mL of the sample solution in 5 µg/mL, 10 µg/mL, 25 µg/mL, 50
µg/mL and 100 µg/mL concentrations. These mixtures were incubated for
30 min at room temperature. After this, the absorbance was measured at
517 nm against a blank. The inhibition of DPPH radical was calculated as
follows:
where Abscontrol is the absorbance of DPPH radical + methanol; Abssample is
the absorbance of DPPH radical + extract/standard.
2.7 D-glucose-6-phosphate phosphorylase (from Rat Liver) Assay
The livers of male rats of Wistar strain were exercised. A 10%
homogenate was prepared in 150 mM KCl using Potter Elvejhem glass
homogenizer fitted with Teflon pestle. The homogenate was centrifuged at
1500 rpm for 10 min; supernatant was decanted and used as enzyme
source. The effect of extracts and fractions were studied by pre-incubating
the compound in 1.0 mL reaction system for 15 min and then determining
the residual glucose-6-phosphatase activity according to the method of
Hubscherand West [12]. The 1.0 mL assay system contained 0.3 M citrate
buffer (pH 6.0), 28 mM EDTA, 14 mM NaF, 200 mM glucose-6-phosphate,
and enzyme protein. The mixture was incubated at 37°C for 30 min after
which 1.0 mL of 10% TCA was added. Estimation of inorganic phosphates
(Pi) in protein free supernatant was done according to the method of
Taussky and Shorr [13]. Glucose-6-phosphatase activity was defined as
micromole Pi released per minute per milligram protein.
2.8 Statistical Analysis
The values are expressed as Means±SD. The experiments were repeated
three times. Data were analyzed using one way ANOVA followed by
Dunnett’s test for multiple comparisons using the graph pad prism v5.0
(Graph pad software, inc., USA). P values of less than 0.05 were taken to be
significant in the experiments.
3. Results and Discussion
3.1 Phytochemical Screening
The preliminary phytochemical screening of leaf extracts of SC (Table
1) revealed the presence of various compounds such as phenols, alkaloids,
saponins, steroidal-spanning, flavonoids, fatty acids and coumarins. When
1ml of the methanol or aqueous extract was taken in test tube followed by
a few drops of 10% ferric solution was added. Formation of blue or green
color indicates presence of phenols. For the steroids, 1 mL of extract in
chloroform and few drops of conc. H2SO4 was mixed to form brown ring.
Fluorescence was detected by the UV test (365 nm) for chloroform fraction
which indicate the presence of coumarins. Flavonoids were verified for the
studies chloroform, methanol and residual aqueous fraction with Shinoda
test. In this test to the test solution few magnesium turnings and
concentrated hydrochloric acid was added dropwise pink, scarlet or green
to blue color appears after minutes. The presence of saponins was
confirmed by foam-producing properties of these compounds. They were
identified in methanol and aqueous fractions.
Table 1 Results of phytochemical analysis of leaf extracts of SC
Entry
Phytochemicals
Hexane
fraction
Chloroform
fraction
Methanol
fraction
Aqueous
fraction
1.
Phenols
-
+
+++
++
2.
Saponins
-
-
+++
+++
3.
Steroids
+
+++
+
-
4.
Flavonoids
-
+
+++
+++
5.
Coumarins
+
+++
+
-
6.
Fatty acids
+++
+
-
-
+++: Strong intensity reaction; ++: Medium intensity reaction;
+: Weak intensity reaction; −: Nondetected
3.2 Extraction Yield
Results of extraction yield showed that the amount of extraction crude
of SC depends upon the solvent nature and it varied from 10.2to 41.0%
with a descending order of methanol >water > chloroform > hexane (Table
2). Extraction with methanol resulted in the highest amount of total
extractable compounds, whereas the extraction yield with hexane was
small in comparison to other solvents. Higher extraction yield in methanol
might be due to the fact that it easily penetrates the cellular membrane
and extracts the intracellular ingredients from the plant material. These
results showed that SC contains more of polar compounds than the others.
3.3 Total Phenolic Content
The quantitative determination of total phenol was determined with the
Folin-Ciocalteu reagent. The total phenols were expressed as mg/g Gallic
acid equivalent using the standard curve of gallic acid (Fig. 1). Linearity of
calibration curve was achieved between 1 to 8 µg/mL and calculated as an
equation: y = 0.0837x-0.0513, R2= 0.999, where y is absorbance at 760 nm
and x is total phenolic content in the extracts and fractions. The maximum
phenolic content was found in the methanol fractions (86.2 ± 2.6). In the
hexane and chloroform fractions, phenolic compounds could not be
detected. These results demonstrate clearly that the content of phenolic
compounds is dependent on the polarity of the solvent used; higher the
polarity of the solvent, higher the content of phenolic compounds.
Moreover, SC leaves can be considered as a good source of phenolic
compound.
Fig. 1 Standard curve of Gallic acid
Table 2 Extraction yield and Total phenolic content of SC leaves extracts
S. No.
Solvent
Extraction yield
(%w/w)
Total phenolic content (mg
GAE/g extract)
1
Ethanol
10.0 ± 1.5a
81.8 ± 3.4a
2
Hexane
10.2 ± 0.5b
ND
3
Chloroform
25.4 ± 2.1c
21.9± 1.8b
4
Methanol
41.0 ± 1.8d
86.2 ± 2.6c
5
Water
16.4 ± 2.4e
ND
Values are mean ± standard deviation of triplicate experiments. Different letters in
columns show significant differences at p < 0.05; ND, not detected

136
Tanveer Ahamad et al. / Journal of Natural Products and Resources 3(2) (2017) 134–136
Cite this Article as: Tanveer Ahamad, Devendra Singh Negi, Mohammad Faheem Khan, Phytochemical analysis, total phenolic content, antioxidant and an tidiabetic activity of Sansevieria cylindrica leaves
extract, J. Nat. Prod. Resour. 3(2) (2017) 134-136.
3.4 Antioxidant Activity
Most of the methods of determination of total antioxidant activity
characterize the ability of the tested compound or product to scavenge
free radicals where DPPH is the best example to measure the free radical
scavenging activity. It is recommended for studies with electron and
hydrogen donating compounds such as phenols or flavonoids. Our result
showed that the methanol fraction has the highest percentage of phenol
content. Keeping in view, we evaluated the antioxidant activity of the
ethanol extract and its fractions viz hexane, chloroform, methanol and
aqueous by determining the percentage inhibition of DPPH radical.
Ascorbic acid was used as standard for the present investigation. A
significant inhibition of DPPH free radical was observed in ethanol extract
and methanol fraction at the concentration of 100 μg/mL (Table 3).
Ethanol extract inhibits 80.5% of the DPPH free radical at 100 μg/mL,
whereas methanol fraction showed 83.6% inhibition of DPPH radicals at
the same concentration (Fig. 2). These results were compared with
ascorbic acid as standard drug which shows 92.1% inhibitory activity. In
addition, hexane and chloroform fraction were also evaluated for
inhibition of DPPH radical, but they have been found to show weak
inhibitory activity.
Fig. 2 In vitro antioxidant activity of ethanol extract and methano l fraction at
different concentration
Table 3 DPPH assay showing % inhibition of the ethanol extract and its fractions
S.No.
Concentration
(μg/mL)
% inhibition of free radicals
Ethanol
extract
Hexane
fraction
Chloroform
fraction
Methanol
fraction
Aqueous
fraction
1
5
62.8
20.1
25.6
68.3
ND
2
10
73.1
21.7
27.4
76.6
ND
3
25
75.1
21.9
30.8
80.2
ND
4
50
76.2
24.5
32.7
78.5
ND
5
100
80.5
29.3
35.1
83.6
ND
ND: not determine
3.5 Antidiabetic Activity
Increased oxidative stress is involved in diabetes by the generation of
oxygen derived free radicals. The generation of free radicals may lead to
lipid peroxidation in diabetes mellitus by glucose degradation, non-
enzymatic glycation of proteins and the subsequent oxidative degradation
[14]. Glucose 6-phosphatase is as hydrolyzing enzyme and plays a key role
in the homeostatic regulation of blood glucose levels via formation of a
phosphate group and a free glucose on hydrolysis [15]. Ethanol extract and
all the fractions were evaluated for glucose-6-phaphatase inhibitory
activity against standard drug sodium orthovanadate. The ethanol extract
showed 51.3% inhibitory effect (Table 4). Furthermore, the methanol
fraction exhibited 57.9% inhibition of glucose-6-phosphatase enzyme at
100 µM concentration which was most active in comparison to other
fractions such as hexane, chloroform and aqueous fraction.
Table 4 In-vitro glucose-6-phosphatase enzyme inhibition results of extract and
fraction at 100 μM concentration
S.No.
% inhibition of glucose-6-phosphatase enzyme
Ethanol
extract
Hexane
fraction
Chloroform
fraction
Methanol
fraction
Aqueous
fraction
1
51.3
25.1
33.5
57.9
ND
Percentage inhibition of sodium orthovanadate is 53.4% at 100 μM concentration
4. Conclusion
In conclusion, our results show the presence of various phytochemicals
in fractions of plant which may be responsible for the pharmacological
activity. Total phenolic content, antioxidant and antidiabetic activity were
also discussed. Methanol fraction has highest phenolic content that is 86.2
± 2.6. Ethanol extract and methanol fraction showed maximum DPPH
radical scavenging activity and they inhibited 80.5% and 83.6% free
radicals at 100 μg/mL concentration, respectively. In order to evaluate the
antidiabetic activity, methanol fraction showed 55.5% inhibition of
glucose-6-phosphatase enzyme. Extracts of SC appear to be attractive
materials for further studies leading to possible drug development for
antioxidant and diabetes, which is relatively inexpensive, less time
consuming and more economical for drug development and discovery.
Acknowledgements
Authors are highly grateful to EET (Era’s Educational Trust), Era’s
Lucknow Medical College & Hospital, Lucknow, UP for financial assistance
to carry out this work.
References
[1] B.B. Petrovska, Historical review of medicinal plants’ usage, Pharmacogn Rev.
6 (2012) 1-5.
[2] F. Firenzuoli, L. Gori, Herbal medicine today: Clinical and research issues, Evid.
Based Complement Alternat. Med. 4 (2007) 37-40.
[3] M. Ekor, The growing use of herbal medicines: issues relating to adverse
reactions and challenges in monitoring safety, Front. Pharmacol. 4 (2013) 177-
187.
[4] J.W. Li, J.C. Vederas, Drug discovery and natural products: End of an era or an
endless frontier, Science 325 (2009) 161–165.
[5] M.S. Brewer, Natural Antioxidants: sources, compounds, mechanisms of action,
and potential applications, Compr. Rev. Food Sci. Food Saf. 10 (2011) 221-247.
[6] R. Takawira, I. Nordal, The genus of Sansevieria (family Dracaenaceae) in
Zimbabwe, Acta. Hortic. 552 (2003) 89-199.
[7] A. Said, E.A. Aboutabl, F.R. Melek, GARA. Jaleel, M. Raslan, Steroidal saponins
and homoisoflavanone f rom the aerial parts of Sansevieria cylindrica Bojer ex
Hook, Phytochem. Lett. 12 (2015) 113-118.
[8] A.A. Da Silva, B.P. Da Silva, J.P. Parente, A.P. Valente, A new bioactive steroidal
saponin from Sansevieria cylindrical, Phytother. Res. 17 (2003) 179–182.
[9] G.E. Trease, W.C. Evans, Pharmacology 11th Ed., Brailliar Tiridel and Macmillian
Publishers, London, 1989.
[10] J.Y. Lin, C.Y. Tang, Determination of total phenolic contents in selected fruits
and vegetables as well as their stimulatory effect on mouse splenocyte
proliferation, Food Chem. 101 (2007) 140-147.
[11] R.P. Singh, K.N.C. Murthy, G.K. Jayaprakash, Studies o n the antioxidant
polyphenol content of aqueous extract from pomegranate peel and seed
extracts using in vitro models, J. Agric. Food. Chem. 50 (2002) 86-89.
[12] G. Hubscher, G.R. West, Studies on the fractionation of mucosal homogenates
from the small intestine, Nature 205 (1965) 799-800.
[13] H.H. Tussky, E. Shorr, A micro colorimetric method for the determination of
inorganic phosphate, J. Biol. Chem. 202 (1953) 675-685.
[14] R.P. Gutierrez, E.G. Baez, Evaluation of antidiabetic, antioxidant and
antiglycating activities of the Eysenhardtia poylstachya, Phcog. Mag. 10 (2014)
404-418.
[15] A. Ghosh, J.J. Shieh, C.J. Pan, M.S. Sun, J.Y. Chou, The catalytic center of glucose -
6-phosphatase. HIS176 is the n ucleophile forming the phosphohistidine-
enzyme intermediate during catalysis, J. Biol. Chem. 277 (2002) 32837-32842.