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European Journal of Applied Sciences – Vol. 10, No. 1
Publication Date: February 25, 2022
DOI:10.14738/aivp.101.11689. Egbunefu, C. O., Iyama, W. A., & Gbode, Y. L. (2022). Phytochemical Evaluation and Characterization of Methanol Extracts from the
Stem-back of Jatropha curcas Plant. European Journal of Applied Sciences, 10(1). 324-332.
Services for Science and Education – United Kingdom
Phytochemical Evaluation and Characterization of Methanol
Extracts from the Stem-back of Jatropha curcas Plant
Egbunefu, Chukwudi Omeni
Rivers State College of Health Science and Management Technology
Oro-Owo, Rumueme, Port Harcourt
Iyama, William Azuka
Rivers State University, Institute of Geosciences and
Environmental Management, Port Harcourt
Gbode, Yenor Lekia
Rivers State College of Health Science and Management Technology
Oro-Owo, Rumueme, Port Harcourt
ABSTRACT
The phytochemical screening and characterization of the alkaloid and glycosides
fractions of methanol extracts of stem-back of Jatropha Curcas plant were carried
out. The phytochemical screening showed that the stem-back had 0.62 ± 0.18%
flavonoids, 0.52 ± 0.42% tannin, 6.99 ± 0.37% Saponin, 28.78 ± 0.39mg/100g
alkaloid and 51.92 ± 0.35mg/100g glycosides. Furthermore, two fractions of these
phytochemicals (i.e. the alkaqloid and glycoside) were characterized using
standard methods. The alkaloid fraction was found to contain curcin (0.44 ±
0.20mg/100g), tetramethylpyrazine (1.22 ± 0.12mg/100g), atropin (2.10 ±
0.32mg/100g), diphenoxylate (0.81 ± 0.32mg/100g), curcarcycline A (0.49 ±
0.17mg/100g) and curcain (0.80 ± 0.20mg/100g). The glycoside fraction contained
linamarin (0.25 ± 0.09mg/100g), lotaustralin (2.14 ± 0.09mg/100g), prunasin (0.40
± 0.12mg/100g), dhurrin (2.49 ± 0.32mg/100g), amygdelin (15.90 ± 0.55mg/100g),
quabain (3.98 ± 0.45mg/100g), digitoxin (13.69 ± 0.40mg/100g), digitalis (4.14 ±
0.25mg/100g) and dioxin (8.89 ± 0.51mg/100g). These active ingredients found in
this herbal plant are of great importance in the pharmaceutical industry and can be
harnessed to better the lot of humanity more especially in this Covid-19 era when
all hands are on deck to find a possible cure for the pandemic that has ravaged the
entire world.
Keywords: Alkaloids, glycosides, Jatropha curcas, screening, phytochemicals, stem-back
INTRODUCTION
Nature has endowed mankind with all that is required to take care of it on planet earth. The
earth is filled with so many natural resources at man’s disposal. These enormous resources at
man’s disposal includes the vegetation (i.e abundant plant resources) that serves not only for
food but also to take care of the health need of man. Several plants exist with very high nutritive
and medicinal values and yet remain unexploited for human and animal benefits. Plants are
very vital because they are a fundamental part of life on earth, because they generate oxygen,
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Egbunefu, C. O., Iyama, W. A., & Gbode, Y. L. (2022). Phytochemical Evaluation and Characterization of Methanol Extracts from the Stem-back of
Jatropha curcas Plant. European Journal of Applied Sciences, 10(1). 324-332.
URL: http://dx.doi.org/10.14738/aivp.101.11689
food and medicine that allows human and other higher life forms to exists (Izevbige et al.,
2004). In addition, to using green leaves as food, specific green leaves make excellent natural
medicines. They work as Hippocrates, the father of medicine said “let food be your medicine
and medicine be your food” (Izevbige et al., 2004). Plants form the main ingredients of medicine
in traditional system of healing and have been the source of inspiration for several major
pharmaceutical drugs (Obute, 2005). African happens to be one of the continent that is greatly
blessed with different varieties of this natures endowment known as plants, the majority of
which are of medicinal value. But her short coming has been how to fully harness this natures
gift and get the best out of it. The use of medicinal plants is increasing worldwide, in view of the
tremendous expansion of traditional medicine and growing interest in herbal treatments.
Plants are used in medicine to maintain and augment health-physically, mentally and spiritually
as well as to treat specific condition and ailments (Igoli et al., 2005). The plant kingdom
contributed immensely to human health when no synthetic medicines were available, and when
no concept of surgery existed. There is therefore need to conserve these plants associated with
indigenous knowledge for our development and good health (Esimone et al., 2009). One of the
smallest countries in Africa (Madagascar) came to global limelight and popularity this Covid- 19 period for their alleged claim of having been able to come up with herbal remedy that could
cure covid-19 ailment. This claim drew the attention of some Western Countries like Russia and
others who placed order for the claimed herbal remedy. Unfortunately, for them, the clinical
trials conducted on this herbal remedy by these Western nations could not proof its efficacy,
which is an indication of work not properly done. This fact does not therefore rule out the
efficacy of herbal remedies. It is in the light of these, that this study, Phytochemical evaluation
of extract of stem-bark of this medicinal plant, Jatropha curcas is carried to ascertain the active
indigents(Phyto-chemicals) in its stem-bark that could be of great benefits to the
pharmaceutical industries in formulating or compounding remedies (drugs) that could be of
great use in treatment of ailments, Covid-19 not excluded.
MATERIALS AND METHODS
The Jafropha curcas stem-bark used for the work was obtained in one of the local communities
(Egwi) in Etche Local Government Area of Rivers State, Nigeria. It is a life tree that is used for
medicinal purpose. The stem-bark was sun-dried and pulverized and stored in an air-light
container for further use (Nwala et al., 2013).
Preparation of Extract
Abort 250g of the pulverized stem-bark sample was extracted with methanol. The extract was
filtered using whatman’s No 1 filter paper and the filtrate was concentrated to dryness in a
vacuo using a rotary evaporator to remove the methanol (Nwala, et al.,2013).
Phytochemical Screening Test for the Extract
A small portion of the dry extract was subjected to the phytochemical test using methods to test
for tannins, saponins, flavonoids, alkaloids and glycosides (Makkor & Becker,1999; Hodek,
et.al.,2002; Li et al., 2003; Akinpelu et al.,2009; Kaur & Arora, 2009; Nwala et al., 2013; Igbinosa,
et al.,2009).
Test for Tannins
About 1g of the sample was put into 250ml conical flask containing 100ml of distilled water and
the flask was boiled for 1hour and then filtered through a whatman (No 541) filter paper (While
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European Journal of Applied Sciences (EJAS) Vol. 10, Issue 1, February-2022
Services for Science and Education – United Kingdom
Warm) into a volumetric flask. The sample solution was hence made up to 100ml after cooling
at room temperature.
Tannic acids standard solution was freshly prepared by dissolving 0.01g of tannic acid in
distilled water diluting to 100ml (1.omg/ml). Zero to 10ml ()-10ml) tannic acids standard
solution was pipette into 100ml volumetric flask to give standard solution of concentration
varying from 0.00-10mg tannic acid. Ten milliliters (10ml) of the sample wee then pipetted into
100ml volumetric flask. Then 5ml of folin-Denis reagent and 20ml 0f 17% (W/V) sodium
carbonate solution were added to both the standard and sample solutions. The solutions were
then diluted to the 100ml mark of the volumetric flask with distilled water and mixed. They
were kept in a water bath at 370C for 20 minutes. Their absorbances were measured
spectrophotometrically at 760nm. The value of the tannin content of the sample was
extrapolated from the standard graph as mg tannin/100g sample.
Saponin Content Determination
One green (1g) of the sample was folded in a filter paper (in a thimble) and put in a socket
extractor and a refuse condenser filtered on top. Extraction was done with acetone in 250ml
capacity round bottomed flask for 3 hours. The apparatus was dismantled and a 15ml capacity
round bottom flask containing 100cm3 of methanol was filtered to the extractor and another
extraction was carried out for 3 hours with methanol. The methanol was recovered by
distillation and the flask oven dried and weighted. The second extraction was done in order to
maintain the change in weight at the end of the second extraction and the saponin content was
then calculated.
Determination of Flavonoids
Ten grams of the sample was extracted in a clinical flask with a rubber lip placed in an ultrasonic
bath with 100ml Ethanol - water (80;20, v/v) solution at 25oCfor 15 minutes repeated 3 times.
The extract was isolated by membrane filtration and them transferred to a rotatory evaporator
devices and concentrated under vacuum until the residual volume was 5lm; the residual was
washed by Ethyl acetated successively followed by filtering and drying, the flavoniod was then
calculated.
Determination of Alkaloids
Two hundred (200ml) of 20% acericacide in ethanol was added to 5g powered sample, covered
and allowed to stand for 4hours. The filtrate was then concentrated on a water bath to 1/4 of
its original volume. Concentrated ammonium hydroxide was added drop wise to the extract
until the precipitation was complete. The whole solution was allowed to settle; collected
precipitation we're washed with diluted ammonium hydroxide and then filtered. The reside
due was, weight and expressed as the Alkaloids.
Determination of Glycosides
One from (IG) of the smoke was weight into 800ml round bottom flask and 200ml of distilled
water added to the flask and allowed to stand for 24hrs. The solution was then distilled to
150ml of distillate, were added 8ml of 6M ammonium hydroxide (NH4OH) and 2ml of 5%
potassium iodide (KI). The resultant solution was titrated against 0.02N silver nitrate (AgNO3).
The titre value was multiplied by 1.08 mg HCN. One (1) ml of 0.02NAgNO3 being equivalent to
1.08mg HCN was used and the glycoside content was then calculated.