J. Chem Soc. Nigeria, Vol. 45, No.

2, pp 324 - 336 [2020]

CHEMICAL CONSTITUENTS AND PHARMACOLOGICAL USE OF ASYSTASIA GANGETICA

(CHINESE VIOLET) AS AN ANTI-ULCER PLANT
I.O Eriamiatoe 1*, M.O Edema 2, T. Eriamiatoe 3 G.E Okpara4
1
Department of Chemistry, University of Benin, Benin City, Nigeria
2
Department of Chemistry, Federal University of Petroleum Resources, Effurun, Delta State, Nigeria.
3 4
Laboratory Technologist Department of Chemistry, University of Benin, Benin, Nigeria, Chemistry
Department University of Benin, Nigeria
*Correspondents Author: isiokem@gmail.com, +2347060498473
Received 10 December 2019; accepted 29 January 2020, published online 29 March 2020

ABSTRACT
The whole plant extract of Asystasia gangetica (Chinese violet) was investigated for chemical
constituents and anti-ulcer activity. The test animal used was albino/wister rat. Asystasia gangetica
(Chinese violet) is used in folklore medicine for various ailments like asthma, rheumatism and pains etc.
Toxicity of the extract was determined using Dietrich„s method. The ability of the methanolic extract of
Asystasia gangetica in preventing and treatment of ethanol-induced ulcer model in rats was investigated.
The extract was prepared by Soxhlet Extraction with 80% methanol. The resultant extract of methanol
was concentrated by removing the solvent using a rotary evaporator and stored in a refrigerator at 4 oC. A
yield of 18.6% w/w dry extract was obtained. The acute toxicity of the methanolic A.G extract showed no
death or observable sign of toxicity at a dosage of 4000mg/kg given per os after 24hrs and to 14 days with
10g/kg body weight of the test animals. Using the ethanol induced ulcer model, 125mg/kg; 250mg/kg and
500mg/kg doses of the plant extract gave an ulcer index and percentage ulcer inhibition of (2.20±0.96,
42.61%), (1.60±0.67, 64.35%) and (0.20±0.18, 89.56%) respectively as compared to the positive control
of 30mg/kg of 20mg/kg omeprazole that gave (4.00±0.63, 29.56%). The result showed a dose dependent
activity for the plant extract, as concentration of the plant extract increases the percentage ulcer inhibition
increases. Asystasia gangetica extract (AGE) exhibited a more gastro-protective effect against ethanol-
induced stomach ulcers at 125, 250 and 500 mg/kg than omeprazole (20 mg/kg) and solvent treated
(control) rats. Major constituents detected by GC-MS were octadecenoic acid ester (RT: 26.55; 8.26%),
an unsaturated fatty acid ester, squalene (RT: 31.82; 1.84%), a cholesterol precursor and beta-sitosterol
(RT: 32.46: 3.78%). The IR bands of the alkaloid fraction were 1380.11cm-1 (C-H) band of methyl,
1652.09cm-1 (N-H band) of amine and 2929.97cm-1 (O-H stretch) of carboxylic acid. The presences of
these compounds in the extract fraction suggest that the plant has rich medicinal value and hence, the
preventive property of A. gangetica justified its ethno medical use as anti-ulcer by patients.
INTRODUCTION by prescription, severe stress (e.g.: trauma,
burns), alcohol, smoking, bile reflux, pancreatic
Peptic ulcer disease (PUD) is a benign lesion enzyme reflux and radiation (7) and if not
which occurs in the stomach or duodenum where properly treated, ulcers can lead to serious health
the mucosal epithelium is exposed to acid and problems, including: bleeding, perforation,
pepsin (1, 2, 3). It develops when the delicate gastric outlet obstruction from swelling or
balance between some gastro protective scarring that blocks the passageway leading
aggressive factors is lost (4; 5; 6). No single from the stomach to the small intestine (6). Most
caused has been found for ulcers. Ulcers can be antiulcer drugs require prolong period of intake,
caused by infection with a type of bacteria yet ulcer relapse is a common occurrence (7 )
called Helicobacter pylori (H. pylori), use of and many of these antiulcer drugs have various
painkillers called non-steroidal anti- adverse effects (8). Over the years, plant and
inflammatory drugs (NSAIDs), such as aspirin, plant products have been used for treatment of
naproxen, ibuprofen and many others available
 J. Chem Soc. Nigeria, Vol. 45, No. 2, pp 324 - 336 [2020]
several ailments and diseases and even now,
medicinal plants is becoming a more viable
alternative treatment over synthetic drugs on
peptic ulcer management/ treatment due to its
lower cost, effectiveness and accessibility as
well as little or no side effects.
Asystasia gangetica is a member of the
Acanthaceae family. It is a perennial
herbaceous, ascending and branching plant.
Flowers are pale purple blue to violet or lime
white in colour(9). Report on phytochemical
screening of the plants extract showed the
presence of carbohydrates, protein, alkaloids,
steroidal aglycones, saponins, flavonoids,
reducing sugars and tritepenoids (10). The plant
has been claimed to have anti-inflammatory,
anti-diabetic, anti-asthmatic, analgesic and
bronchospasmolytic (11; 12; 13). In India, the
whole plant is used as juice for treating
rheumatism and the root paste for skin allergies
(13). In Nigeria the extract of the whole plant is
given to women in labour to ease the pain during
childbirth. It is also given to pregnant women
with constipation in late pregnancy (12). This
research was aimed to determine the chemical
constituents and the pharmacological use of
Asystasia gangetica as an anti-ulcer agent in
order to provide a scientific validation of its use.
2.0 MATERIALS AND METHODS
2.1 Plant Material
The whole plants of AG were collected from the
environment of university of Benin. It was
Identified and authenticated in the Department
of Plant Biology and Biotechnology. A voucher
specimen (No. UBH A460) has been deposited
for future reference.
2.2 Preparation of aqueous extract
The fresh plant samples were air dried in the
laboratory for about five weeks and then milled
with a milling machine. The mass of the crushed
sample was found to be 1200g. The continuous
soxhlet extractor apparatus was used as the
chemical method for obtaining the organic
extract from dried powdered sample of Asystasia
gangetica plant using methanol solvent. 46g of
solid sample was placed in a porous thimble in
the chamber, extracting solvent in the boiling
flash below. The resultant extract of methanol
was concentrated by removing the solvent using
a rotary evaporator and stored in a refrigerator at
4oC. The concentration and percentage yield of
the extract were determined.
2.3 Phytochemical analysis
The preliminary phytochemical analysis of A.
gangetica aqueous extract was carried out for
glycosides, carbohydrates, saponins, flavonoids,
triterpenoids, tannins and alkaloids. It was
performed using standard procedures by
Sofowora (1993), Trease and Evans (1989), as
well as Odebiyi and Sofowora (1978). (14,
15,16).
2.4 Isolation of Alkaloids
50ml of diethyl ether was used as a solvent to
dissolve the plant extract in a 100ml separating
funnel. 20ml of 2M HCl was added, shaken and
allowed to separate. The lower aqueous layer
was run-off into a conical flask. The extraction
was repeated with 2 more portions of 20ml of
2M HCl. The acidic fraction was then
neutralized to pH 7 using aqueous solution of
Na2CO3. The organic compound was then
separated from the aqueous mixture by
extracting with diethyl ether. The ether was
evaporated to obtain the isolate.
2.5 TLC and GC-MS analysis
The thin-layer chromatography was carried out
on the extract of Asystasia gangetica and Gas
chromatography- mass spectrometric (GC-MS)
analysis was carried out on a GC- mass
spectrometer filled with an HP-5 MS (5%
phenysiloxane) column at a temperature
programme of 70oC (2 minutes) increase at
10oC/ min to 280oC and held for 7 minutes. The
carrier gas was nitrogen and flow rate,
1.80mL/min.
2.6 FT-IR Spectroscopic analysis
The extracted Asystasia gangetica alkaloid
isolate was analysed by Fourier transform IR
(FTIR) instrument. using KBr.
2.7 Test animals
 J. Chem Soc. Nigeria, Vol. 45, No. 2, pp 324 - 336 [2020]
Wistar/ albino rat of both sexes weighing the
range 140-230g, purchased from the animal
house, Department of Pharmacology, University
of Benin, kept in multiple cages at room
temperature and maintained on standard diet
(Grower mash Feeds Ltd, UNIBEN, Edo State)
and water ad libitum for four weeks before
commencement of the experiments were used.
They were fasted for 24 hours, with free access
to water prior to the experiments. Ethical rules
guiding the use of animals for experimentation
were strictly adhered to.
2.8 Acute oral toxicity test
Thirty (30) matured albino Wistar rats of both
sexes were marked with 10% picric acid,
weighed and randomly separated into 5 groups
(A – E) with each group having 5 rats. Range
finding studies were done to determine the doses
and these rats were administered orally with
1000, 2000 and 5000mg/kg of the extract and 5
% v/v Tween-80 in water (negative control),
respectively. They were observed within 24 h
for any death or observable signs of toxicity and
for 14 days thereafter (17).
2.9 Ethanol - induced ulcers
Twenty-five rats treated as given above and
divided into cage (1– 5) of 5 animals each
labeled A-E, were administered orally with 5 %
v/v of tween-80 in water (negative control), 125,
250 and 500 mg/kg of the extract, and 30 mg/kg
omeprazole (positive control), respectively. One
hour thereafter, 1 ml of absolute ethanol was
administered (p. o.) to all the rats. Another 1 h
later, the animals were sacrificed after
chloroform anaesthesia, their stomach isolated,
opened along the greater curvature, and gently
washed in distilled water. They were spread out
on clean sheets of white cardboard paper, held in
position with the aid of office pins.
The ulcer lesions were counted using a
magnifying glass and their diameters were
measured using a straight rule and scored on a
scale of 0 - 10 (17) as follows:
Score 0: No ulcer; 1: haemorrhagic and slightly
dispersed ulcers, diameter < 2 mm; 2: one ulcer
as described above, diameter up to 5 mm; 3:
more than one ulcer, diameters up to 5 mm; 4:
one ulcer, diameter > 5 mm; 5: more than one
ulcer, diameters > 5 mm; 10: total ulceration and
haemorrhage.
The ulcer index (UI) was calculated thus:
UI = UN + Us + Up x 10-1.
Where UN= Average number of ulcers per
animal; US = Average of severity score; Up =
Percentage of animals with ulcers.
Percentage ulcer inhibition was calculated as:
UIC
– UIT X 100
UIC
UIC: Ulcer index of control; UIT: Ulcer index of
test.
Statistical analysis
Statistical analysis was done using student„s t-
test and significance of difference was accepted
at p < 0.05. Data are presented as mean ±
standard error of mean (SEM).
3.0 RESULTS
3.1 Percentage Yield Of Methanol Extract of
Asyatasia gangetica.
Total Mass of the ground plant = 241.9g
Mass of concentrated extract (methanol) =
44.99g
Therefore, % yield = 44.99 x 100
= 18.6w/w
241.9
3.2 Taste of Sample
The taste of the whole plant was found to be
tasteless.
3.3 PHYTOCHEMICAL SCREENING
 J. Chem Soc. Nigeria, Vol. 45, No. 2, pp 324 - 336 [2020]

TABLE 1A: Chemical Tests on Asystasia ed

gangetica Methanol Extract
Dark Alkalo
Expect Metha ALKALI Reddish ids
brown
ed nol ODS brown presen
precipi
Name of Observ extrac infere Dragendo precipit tate t
S/N Test ation t nce ff‟s ate observ
Pale reagent ed
yellow Alkalo
Reddish brown ids
PHENO colour Largel precipi
LIC ation y brown presen
precipit tate t
1COMPO Pale observ presen observ
.UND yellow ed t Wagner‟s ate
reagent ed
Brown Glyco light Alkalo
2 ring sides Yellow yellow ids
.GLYCO Brown observ presen 9Picric precipit precipi presen
SIDES ring ed t .reagent ate tate t
Frothi Saponi
Persiste ng ns
3SAPONI nce observ presen 3.3 TOXICITY
.N frothing ed t
The acute toxicity study of A. gangetica whole
Yello plant methanolic extract (up to 10 g/kg) showed
w no death nor was there any observable sign or
precipi Flavon manifestation of toxicity after 24 h and for 14
Yellow tate oids days thereafter. This indicated that the plant
4FLAVO colourat observ presen may be relatively safe at the tested doses.
.NOID ion ed t
An intense and widespread thickened gastric
Reddis lesion of the mucosa was evident in the control
h rats that received 1ml of absolute ethanol.
Bluish brown Tanni Ethanol is found in various alcoholic beverages
5 precipit precipi ns and drinks consumed by about 45 – 50 % of the
.Tannin ate tate absent Nigerian population (19). Hence, ethanol was
used as the test model to determine any
Green usefulness of this plant in preventing this. (18)
Pale precipi
yellow tate Eugen TABLE 2A: Weight of Each Rat in Each of
6 precipit observ ol the Labeled Cages
.Eugenol ate ed absent CAGES A(g) B(g) C(g) D(g) E(g)
Blue or Blue Steroi 1 169.3 201.9 142.3 169.5 208.0
green colour ds 2 226.1 162.3 146.5 200.0 180.9
7 colourat observ presen 3 212.3 201.6 169.3 201.9 200.0
.Steriod ion ed t 4 226.4 216.8 201.6 212.0 191.5
5 204.2 220.0 210.1 223.0 216.0
Reddish Reddis Terpe
brown h nes
8 colourat brown presen Cage one was used as the negative test
.Terpenes ion colour t control which was given 10ml/kg of 5%
observ tween 80.
 J. Chem Soc. Nigeria, Vol. 45, No. 2, pp 324 - 336 [2020]

Cage two was used as the positive test Therefore Rat A will be administered 1.36ml of
control which was given 20mg/kg 20mg/kg omeprazole solution.
omeprazole.
This calculation was repeated for other rats in
Cage three was given 125mg/kg of cage 2.
concentrated methanolic plant extract
TABLE 2C: Doses of 30mg/kg of 20mg/kg
Cage four was given 250mg/kg of omeprazole administered to the labeled Rats
concentrated methanolic plant extract in Cage 2
Cage five was given 500mg/kg of Cage 2 A B C D E
concentrated methanolic plant extract Ml 1.4 1.0 0.9 1.2 1.1
Calculations for cage 1:
Calculations for cage 3:
For Rat A=
For Rat A=
2.5ml of tween 80 was made to mark with
distilled water in 50ml standard flask. 450mg/kg of concentrated methanolic plant
extract was dissolved in 15ml of 5% Tween-80
If 1000g Rat get 10ml of 5% tween-80, therefore
solution. Therefore 30mg/kg of concentrated
means that 169.3g Rat will be administered Xml
methanolic plant extract will be dissolved in
Rat A: Xml = 169.3g x 10 = 1.693ml 1ml.
approximately 1.7ml
450 x 1 = 30mg/kg
1000
15
TABLE 2B: Doses of 10ml/kg of 5% tween 80
If 1000g Rat is administered 125mg/kg of
administered to the labeled Rats in Cage 1
concentrated methanolic plant extract therefore
Cage A(ml) B(ml) C(ml) D(ml) E(ml) 212.3g Rat will be administered = 212.3 x 125
1 = 26.53mg/kg. 1000
1.7 2.0 1.4 1.7 2.1 If 30mg/kg will be dissolved in 1ml, therefore
26.53mg/kg will be dissolved in
Calculations for cage 2: 26.53 x 1= 0.89ml.
For Rat A= 30
20mg/kg omeprazole was dissolved in 4ml Therefore Rat A will be administered 0.89ml
Tween-80. concentrated methanolic plant extract solution.
If 1000g Rat is administered 30mg/kg of TABLE 2D: Doses of 125mg/kg of
20mg/kg omeprazole therefore concentrated methanolic plant extract
226.1g Rat will be administered 226.1 x 30 = administered to the labelled Rats in Cage 3
6.783mg/kg, Cage 3 A B C D E
1000 Ml 0.9 0.8 0.7 0.8 0.8

If 20mg/kg is dissolve in 4ml, therefore means

that 6.783mg/kg will be dissolved in Calculations for cage 4:

6.783 x 4 = 1.36ml. For Rat A=

20 450mg/kg of concentrated methanolic plant

extract was dissolved in 15ml of 5% Tween-80
 J. Chem Soc. Nigeria, Vol. 45, No. 2, pp 324 - 336 [2020]

solution. Therefore 30mg/kg of concentrated 102.1 x 1 = 3.40ml.

methanolic plant extract will be dissolved in
1ml. 30

450 x 1 = 30mg/kg Therefore Rat A will be administered 3.40ml

concentrated methanolic plant extract solution.
15
TABLE 2F: Doses of 500mg/kg of
If 1000g Rat is administered 125mg/kg of concentrated methanolic plant extract
concentrated methanolic plant extract therefore administered to the labelled Rats in Cage 5
226.4g Rat will be administered = 226.4x 250
= 56.6mg/kg. Cage 5 A B C D E
Ml 3.4 3.7 3.5 3.7 3.6
1000
If 30mg/kg will be dissolved in 1ml, therefore Table 2g: Results Of Measurement Of Ulcer
26.53mg/kg will be dissolved in And Scores of Ulcer
56.6 x 1 = 1.89ml. RAT MEASUREMENT SCORE
LABELLED OF LENGTH OF
30
IN CAGE 1 (mm) ULCER
Therefore Rat A will be administered 1.89ml
concentrated methanolic plant extract solution. A 0.6, 0.5, 0.1, 0.1, 1
0.6, 0.7, 0.1, 0.2, 0.7
TABLE 2E: Doses of 250mg/kg of
concentrated methanolic plant extract B 0.4, 0.2, 0.5, 1.2, 1
administered to the labeled Rats in Cage 4 0.3, 0.7, 0.5, 0.6,
0.7, 0.1, 0.1, 0.2,
Cage 4 A B C D E 0.7, 0.5, 0.1, 0.3
Ml 1.9 1.8 1.7 1.8 1.6
C 0.7, 0.5, 0.6, 0.7, 1
0.1, 0.1, 0.2, 0.7,
Calculations for cage 5: 0.5, 0.1, 0.3, 0.2
For Rat A= D 0.6, 0.5, 0.1, 0.1, 1
450mg/kg of concentrated methanolic plant 0.6, 0.1, 0.2, 0.7,
0.5, 0.1, 0.3
extract was dissolved in 15ml of 5% Tween-80
solution. Therefore 30mg/kg of concentrated E 0.4, 0.2, 0.5, 1.2, 1
methanolic plant extract will be dissolved in 0.3, 0.7, 0.5, 0.6,
1ml. 0.7, 0.1, 0.1, 0.2,
450 x 1 = 30mg/kg 0.7, 0.5, 0.1

15 RAT MEASUREMENT SCORE

LABELLED OF LENGTH OF
IN CAGE 2 (MM) ULCER
If 1000g Rat is administered 125mg/kg of A 1.0, 0.1, 0.1, 0.2, 1
concentrated methanolic plant extract therefore 0.5
204.2g Rat will be administered = 204.2x 500
= 102.1mg/kg. B 0.4, 0.2, 0.5, 1.2, 2
0.7, 0.5, 0.6, 0.7,
1000 0.1, 0.1, 0.2, 0.7.
If 30mg/kg will be dissolved in 1ml, therefore C 0.7, 0.5, 0.6, 0.7, 1
102.1mg/kg will be dissolved in
 J. Chem Soc. Nigeria, Vol. 45, No. 2, pp 324 - 336 [2020]

D 0.6, 0.5, 0.1 1 Asystasia 125 2.20 42.61

gangetica ±0.96
E 0.4, 0.2 1
Asystasia 250 1.60 64.35
RAT MEASUREMENT SCORE gangetica ±0.67
LABELLED OF LENGTH OF
IN CAGE 3 (MM) ULCER Asystasia 500 0.20 89.56
gangetica ±0.18
A 0.6, 0.5 1
Omeprazole 30 4.00 29.56
B 0.4, 0.2, 0.5, 0.7, 0.8 1 ±0.63
C 0.5 1 Values are Mean ± S.E.M. p < 0.05,
significantly different from control. p < 0.05,
D 0.6, 0.5 1 significantly different from omeprazole using
E 0.4 1 Students t-test. N = 5 animals.

RAT MEASUREMENT SCORE

LABELLED OF LENGTH OF A
IN CAGE 4 (MM) ULCER

A 0.1, 0.1, 0.1, 0.2 1

B 0.1, 0.3 1
C -- 0
D 0.1, 0.1 1
E -- 0
RAT MEASUREMENT SCORE
LABELLED OF LENGTH OF
IN CAGE 5 (MM) ULCER

A ---- 0
B ---- 0
B
C -- 0
D --- 0
E 0.2 1

Table 2H: Effects of pre-administration of

Asystasia gangetica extract on ethanol- C
induced ulcers.
Extract/drug Dose Ulcer % ulcer
(mg/kg) (mg/kg) index inhibition
Control 0.00 12.00 0.00
±1.02
 J. Chem Soc. Nigeria, Vol. 45, No. 2, pp 324 - 336 [2020]

Table 3B: Rf Value and Colour Reaction of

Methanolic Asystasia gangetica Extract
Using
Solvent System: Chloroform
Solvent Front: 8.4

D Distance
travel by Solvent Rf Colour
Origin sample front value change
0.00 0.6 8.4 0.07 greenish
yellow
0.00 0.1 8.4 0.12 Light
green

E 0.00 2.1 8.4 0.25 faint

yellow
Plate 1: Representative rat stomach
presentation in the ethanol- induced ulcer (A 0.00 2.8 8.4 0.33 Dark
– tween-80 10 ml/kg; B -20mg/kg Omeprazole yellow
30 mg/kg; C– AGE 125 mg/kg; D – AGE 250
mg/kg; and E- AGE 500mg/kg).

3.4 Thin Layer Chromatographic Analysis of Table 3C: Rf Value and Colour Reaction of
Acidic, Basic And Neutral Components Of Methanolic Asystasia gangetica Extract
Asystasia gangetica Using
Table 3A: Rf Value and Colour Reaction of Solvent System: Methanol
Methanolic Asystasia gangetica Extract
Using Solvent Front: 8.4

Solvent System: Ethanol Distance

Solvent Front: 8.4 travel
by Solvent Rf Colour
Distance Origin sample front value change
travel by Solvent Rf Colour
Origin sample front value change 0.00 2.5 8.4 0.30 Pale
yellow
0.00 2.7 8.4 0.32 Pale
yellow 0.00 3.2 8.4 0.38 Light
green
0.00 3.2 8.4 0.38 Light
green 0.00 3.7 8.4 0.44 Dark
yellow
0.00 3.3 8.4 0.39 Dark
yellow 0.00 4.0 8.4 0.48 Dark
green
0.00 3.5 8.4 0.42 Dark
green 0.00 4.1 8.4 0.49 Pale
green
 J. Chem Soc. Nigeria, Vol. 45, No. 2, pp 324 - 336 [2020]

0.00 4.4 8.4 0.52 Light chemical compounds identified in the oil
yellowish fraction are presented in Table 4.
green
Table 4: GC-MS Analysis of alkaloid fraction
0.00 4.8 8.4 0.57 Greenish of methanolic extract of Asystasia gangetica
yellow
Retenti
Pea Area
on Name of
k Percenta
Time Compound
Table 3D: Rf Value and Colour Reaction of No ge (%)
(Rt)
Methanolic Asystasia gangetica Extract Using
1. 6.27 Nonen-1-ol 0.55
Solvent System: Hexane
2. 10.06 2- Dodecenal 1.08
Solvent Front: 8.4
3. 12.63 2- Undecenal 1.69
Origin Sample Solvent Rf Colour
distance front 4. 13.78 3,7- 0.34
dimethylnonane
0.00 0.00 8.4 0.00 Green
5. 18.78 Dodecyl 0.95
trichloroacetate
Table 3E: Rf Value and Colour Reaction of 6. 18.99 2-ethylhexylester 0.83
Methanolic Asystasia Gangetica Extract
Using 7. 20.59 9-octadecene 0.31

Solvent System: Methanol : Ethanol (1:1) 8. 22.44 1-heptadecene 3.57

Solvent Front: 8.4 9. 22.55 Octacosane 1.68

10. 24.86 Henicosan-1-ol 6.89
Distance 11. 24.94 Octadecane 2.19
travel by Solvent Rf Colour
Origin sample front value change 12. 26.55 Ethyloleate ((Z)-9- 8.26
octadecenoic acid
0.00 2.1 8.4 0.25 light ethylester)
yellow
13. 36.84 Tetracosan-1-ol 11.33
0.00 3.0 8.4 0.36 Dark (lignoceryl alcohol)
yellow
14. 27.53 Cis-9-octadecenal 3.46
0.00 3.3 8.4 0.39 Green
15. 27.60 15- 1.51
0.00 3.6 8.4 0.43 Greenish hydroxylpentadeca
yellow noic acid
16 27.80 Cyclopentadecanon 3.22
e
3.5 GC-MS ANALYSIS
17. 28.57 1-docosanol 5.81
GC-MS Analysis
18. 29.90 Cyclhexanecarbonx 1.64
The GC-MS chromatogram of the isolated plant ylic acid, Undec-
extract given in figure 2 showed 26 peaks 10-enyl
indicating from the search list of the chemical
abstract service twenty- six compounds. The
 J. Chem Soc. Nigeria, Vol. 45, No. 2, pp 324 - 336 [2020]

19. 29.40 2,4- 16.41 FIG 2: GC-MS chromatogram alkaloid

bis(dimethylbenzyl fraction of methanolic extract of Asystasia
) phenol gangetica
20. 29.72 Bis(2-ethylhexyl) 13.83 3.6 FT-IR ANALYSIS
phthalate
FT-IR Analysis
21. 30.14 Lignoceryl alcohol 4.80
The GC-MS chromatogram of the isolated plant
22. 30.66 Hexanoic acid, 1.82 extract given in figure 3 showed 10 signals
hexyldecylester indicating the several functional groups that are
present in the oil fraction are presented in Table
23. 21.53 1-octacosanol 0.50 5.
24. 31.82 Squalene 1.84 Table 5: FT-IR Analysis of alkaloid fraction
(2,6,10,14,18,22- of methanolic extract of Asystasia gangetica
tetracosahexane,
2,6,10,15,19,23- S/ Frequen Appeara Band Function
hexylmethyl N cy (cm- nce al group
1
)
25. 32.46 Beta-sitosterol 3.78 1. 1067.64 Short C-O Ether
STRET ROR
26. 33.10 Lignoceryl alcohol 1.71
CH
TOTAL 100 2. 1380.11 Short C-H Methyl-
BAND
3. 1455.34 Strong C-H Alkyl-
BAND
4. 1652.09 Short C=O Amide
stretch
5. 2341.66 Short C≡N Nitrile-
STRET
CH
6. 2929.97 Broad O-H Carboxy
STRET lic acid
CH
7. 3175.90 Broad O-H Carboxy
STRET lic acid
CH

Fig 3: FTIR spectra for alkaloid fraction of

methanolic extract of Asystasia gangetica
 J. Chem Soc. Nigeria, Vol. 45, No. 2, pp 324 - 336 [2020]
4.0 Discussion
The result of the phytochemical screening in
Table 1A indicated the presence of phenolic
compounds, glycosides, saponins, flavonoids,
steroids, terpene and alkaloid. Tannins and
eugenols were not detected. The presence of
alkaloids, saponins, flavonoids among others in
any medicinal plants has been reported to be
sources of useful medicines (19). The thin layer
chromatographic results showed that different
phytochemicals show their presence in the
different solvent that was used and changing the
polarity of the solvent helps to get maximum
information about the phytochemicals present
and many of the phytochemicals were found
using methanol and ethanol extract and some
were also present using chloroform extract and
the methanol and ethanol ratio (1:1) extract. The
thin layer chromatography is usually done for
better identification of bioactive constituents of
plant extract and it was observed that among the
five solvents (chloroform, methanol, ethanol,
hexane, methanol and ethanol ratio (1:1)),
methanol was effective extracting maximum
secondary metabolites. Table 3A- 3D gives us
information of the several Rf values that were
obtained from the different solvent used
indicating the solvent polarity that may also help
in selecting a particular solvent system for
further isolation of any further isolation of any
compound from the plant extracts using
chromatographic and spectroscopic techniques
(20).
The acute toxicity of Asystasia gangetica
methanol extracts in this research show no death
or any observable sign of toxicity after 24hrs
and to 14 days with 10g/kg body weight of the
test animals. The ethanol induced ulcer model
adopted in this work involves 5% v/v of tween –
80 in water (negative control); 125mg/kg;
250mg/kg and 500mg/kg of the plant extract
while positive control group were administered
30mg/kg of 20mg/kg omeprazole. The
measurement of the ulceration in millimeters
indicated the prevalence of ulcer in cage 1
(negative control); for cage 2 (positive control)
which was administered 30mg/kg of 20mg/kg
omeprazole offered a fairly protective gastric
mucosa while for cage 3, 4, 5 which were
administered 125mg/kg, 250mg/kg and
500mg/kg doses respectively, ulceration was
reduced. The ulcer index and percentage ulcer
inhibition was recorded as control (12.00±1.02,
0.00%), 125mg/kg dose of plant extract
(2.20±0.96, 42.61%), 250mg/kg dose of plant
extract (1.60±0.67, 64.35%), 500mg/kg dose of
plant extract (0.20±0.18, 89.56%) and positive
control (4.00±0.63, 29.56%). The result showed
a dose dependent activity for the plant extract, as
concentration of the plant extract increases the
percentage ulcer inhibition increases.
From table 4, major component detected from
the alkaloid fractions of the methanol A.G plant
extract are octadecenoic acid ester (RT: 26.55;
8.26%), an unsaturated fatty acid ester,
tetracosan-1-ol (RT: 36.84; 11.33%), a long
chain alcohol, 2,4- bis (dimethylbenzyl) phenol
(RT: 29.40; 16.41%), lignoceryl alcohol (RT:
30.41; 4.80%), a cholesterol precursor,
Ethyloleate ((Z)-9-octadecenoic acid ethylester)
(RT: 264.86; 8.26%), bis(2-ethylhexyl) phthalate
(RT: 29.72; 13.83%) and beta-sitosterol (RT:
32.46; 3.78%). Hexadecanoic acid, octadecenoic
acid ester, unsaturated fatty acid ester and beta-
sitosterol have been found to show the anti-
inflammatory, antioxidant, anticancer,
antibacterial etc [24, 25, 26].The presence of
these 26 compounds in the GC-MS analysis of
the crude methanol A.G extract fraction plays
vital role in biological fields. These compounds
may be responsible for the antiulcer effect of the
plant extract.
From table 5, the methanol extract of A.
gangetica showed characteristic absorption
bands at 1062.64cm-1 (C-O stretch) of ethers;
1380.11cm-1 (C- H band) of methyl; 1455.34cm
-1
(C-H band) of alkyl groups; 1652.09cm-1
(C=O stretch) of amide and 2929.97cm-1 (O-H
stretch) of carboxylic acid. This result indicates
that the alkaloid fraction could be in form of an
amide salts as confirmed by the C=O stretch at
1652.09cm. The –O-H stretch at 2929.97cm-1
has the ability of hydrogen bonding capacity
may probably indicate the high potential of
methanol extract towards the inhibitory activity
against microorganisms. Thus, the presence of
these functional groups in the extract suggests
that the plant has rich phytoconstitutents that has
 J. Chem Soc. Nigeria, Vol. 45, No. 2, pp 324 - 336 [2020]
medicinal value. This research has corroborated
the traditional use of A. gangetica in the
treatment of ulcer.
5.0 CONCLUSION
The findings from this research have indicated
the presence of important chemical constituents
like alkaloids, flavonoids, phenolic and
saponins. Several plants containing high amount
of saponins and its derivatives has been shown
to possess anti-ulcer activity in ethanol induced
ulcer mode(21) and the protective actions of
saponins are probably due to activation of
mucous membrane protective factor and not
necessary the inhibition of gastric acid secretion
(22). Alkaloids are secondary metabolites that
are known for wide biological activities such as
anti-cholinergic, antitumor, antiulcer,
antimicrobial and anti-inflammatory etc (23).
The anti - ulcer activity revealed that the plant
extract had a dose dependent activity and
compete favourably well with the positive
control ulcer drug used for this work. The anti-
ulcer activity of the whole plant of A.gangetica
can also be linked to the presence of alkaloids,
flavonoids, phenolic and saponins. However,
more research work is needed to purify,
characterised, commercialised the active
ingredients of this Plant extract for the treatment
of gastric ulcer.
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