Anti-Diarrhoeal Potential Of Ethanol Extract of Gouania longipetala Leaves
Subscribe to read and download this work.
ABSTRACT
This study investigated the anti-diarrhoeal potential of Gouania longipetala crude extract and its six different fractions in different anti-diarrhoeal study groups in rats, to establish a scientific basis for its use in traditional medicine as an anti-diarrhoeal. The crude ethanol extract was subjected to phytochemical, gas chromatography-mass spectrometry (GC-MS), molecular docking and FTIR analyses. Acute toxicity evaluation of the crude extract was also done to establish its LD50 value. Its crude extract was evaluated using different doses (200, 400, 600 and 800mg/kg body weight) orally for antidiarrheal activity using castor oil-induced diarrhea, charcoal meal transit time and castor oil-induced enteropooling in different groups of albino rats and was repeated with the fractions. Results of quantitative phytochemical analysis showed the alkaloids to be the most abundant in the crude extract (34.30±0.14 mg/100g) followed by phenols (23.19±0.12 mg/100g). Cardiac glycosides had the least amount (3.89±0.04 mg/100g). Others were saponins (18.10±0.05 mg/100g), steroids (13.74±0.19 mg/100g), flavonoids (16.49 ±1.08 mg/100g), terpenoids (7.45±0.10 mg/100g) and tannins (9.72±0.25 mg/100g). Acute toxicity (LD50) value for the extract was found to be >5000 mg/kg body weight. Antimicrobial effects of the crude extract and that of the most effective fraction (F3) were also evaluated. The activities of the crude extract and fractions at different doses up to 800mg/kg body weight were compared with that of the standard drug, loperamide (0.5 mg/kg). Data were analyzed by the software, statistical package for the Social Sciences (SPSS), Version 18.0. Results obtained showed that the crude extract had significant antimicrobial activity at 200%, eliciting zones of inhibition of 10.20±0.85, 12.00±3.30, 9.60±0.72, and 10.37±1.72 mm against the test isolates Klebsiella pneumonia, Shigella flexneri., Citrobacter feundii and Salmonella paratyphi respectively. In the in vivo anti-diarrhoeal studies, the crude extract at all doses used showed significant (P<0.05) antidiarrhoeal activity evidenced by delay in the onset of diarrhea of up to 56.00±4.36 mins when compared with the control 58.60±3.65. The extract also significantly decreased the distance travelled by the charcoal meal in dose- dependent pattern with activities which compared favourably with that of loperamide (P>0.05). Reduction in the intraluminal fluid accumulation in the castor oil-induced diarrheal model was also observed in all animals treated with the extract. All fractions (F1-F6) at 800mg/kg bodyweight of the leaves of Gouania longipetala exhibited significant (P<0.05) antidairreal activity but fraction 3 (F3) had the highest activities in the castor oil induced diarrhea, enteropooling and charcoal meal transit time. GC-MS spectral analysis of the ethanol extract of Gouania longipetala revealed twenty compounds including spartein, kamferol, oleic acid, 2-tetradecanol, dodecanoic acid, 1-octadecene, dodecanoic acid, 5,6-dehydrolupanine, propanoic acid, 3-chloro methyl ester, luparine, sapogenin A, catechin, flavon-3-ol, anthocyanin, resveratrol, linoelaidic acid, anagyrine, methyl 9,12-heptadecadienoate, baptifoline and ethyl oleate. The biological activities of each compound were discussed in present attempt. The molecular docking assay revealed Sapogenin A to possess an analogous feature with loperamide, which suggests that the antidiarrheal activities of Gouania longipetala could be due to the presence of Sapogenin A. The results of this study showed that Gouania longipetala has a significant (P<0.05) antidiarrhoeal effect and could be seen as a potential source of a new anti-diarrhoeal agent.
TABLE OF CONTENTS
Title Page i
Declaration ii
Certification iii
Dedication iv
Acknowledgements v
Table of Contents vi
List of Tables xi
List of Figures xiii
List of Plates xiv
List of Abbreviations xv
Abstract xvi
CHAPTER ONE: INTRODUCTION 1
1.1 Background of the Study 1
1.2 Aim of the Study 3
1.3 Objectives of the Study 3
1.4 Statement of the Problem 4
1.5 Justification for the Study 4
CHAPTER TWO: LITERATURE REVIEW 6
2.1 The Gastrointestinal Tract 6
2.1.1 Basic functions of the gastrointestinal tract 7
2.1.2 Gastrointestinal motility 7
2.1.2.1 Gastrointestinal secretion and absorption 7
2.1.2.2 Gastrointestinal as a barrier 8
2.1.3 Gastrointestinal pathophysiology 9
2.1.3.1 Gut immune system 10
2.1.4 Flavonoids and gastrointestinal tract 10
2.1.4.1 Gastrointestinal tract and flavonoid metabolism 11
2.1.4.2 Gastrointestinal tract and flavonoids in health and disease 12
2.2 Diagnosis 17
2.2.1 Treatment for ibs 19
2.2.2 Inflammatory bowel diseases 20
2.2.2.1 Pathogenesis of Ulcerative colitis 21
2.2.2.2 Crohn’s disease 22
2.3 Diarrhea: A Disorder of the Gastrointestinal Tract 25
2.3.1 Acute diarrhea 25
2.3.1.1 Epidemiology and etiology 26
2.3.1.2 Pathogenesis of acute diarrhea 26
2.3.1.3 Clinical assessment 27
2.3.1.4 Laboratory evaluation 27
2.4 Management Approaches for Diarrhea 28
2.4.1 Fluid therapy 28
2.4.2 Antidiarrheals 28
2.4.3 Antiemetics 30
2.4.4 Antimicrobials 30
2.4.5 Prevention 30
2.5 Electrolyte Transport Dysbiosis in Diarrhoeal Disease 31
2.6 Mechanisms of Action of Anti-Diarrhoeals 36
2.6.1.1 Mechanism of action of probiotics 37
2.6.2.1 Mechanism of action of antimotility agents 41
2.7.0 The Plant Gouania longipetala 41
2.8 Loperamide (immodium) 44
2.8.1 Mechanism of action of loperamide 48
CHAPTER THREE: MATERIALS AND METHODS 49
3.1 Materials 49
3.1.1 Instruments/equipments 49
3.1.2 Materials 49
3.1.3 Chemicals 50
3.2.1 Collection of plant materials and authentication 51
3.2.2 Preparation of Gouania longipetala leaf extract 51
3.3.1 Animals 52
3.3.2 Microorganisms 52
3.4.1 Qualitative phytochemical screening of the extract 52
3.4.2 Quantitative phytochemical screening of the extract 54
3.5 Gas Chromatography-Mass Spectrometry (GC-MS) Analysis of the Extract 58
3.6 Acute Toxicity (LD50) Evaluation of Gouania longipetala crude Extract 59
3.7 In vivo evaluation of the effect of gouania longipetala extract on
charcoal meal transit in rats 59
3.8 Effect of Gouania longipetala extract on castor oil-induced diarrhoea
in rats 60
3.9 Effect of crude extract of Gouania longipetala on castor oil-induced
fluid accumulation and serum electrolyte concentrations in rats 62
3.9.1 Estimation of serum electrolytes 63
3.10 Bioassay-guided Fractionation of the Ethanol Extract of gouania longipetala
leaf exract 66
3.10.1 Preparation of sample slurry 66
3.10.2 Packing of the glass column 67
3.10.3 Gradient elution solvents 68
3.10.4 Thin- layer chromatography (TLC) 70
3.10.4.1 Plate preparation for thin-layer chromatography 69
3.10.4.2 TLC solvent systems and detection 69
3.11 In vitro evaluation of anti-microbial activity of gouania longipetala leaf
crude extract and fractions 73
3.11.1 Preparation of stock solution of gouania longipetala leaf crude extract
and most bioactive fraction 73
3.11.2 Reactivation of stock cultures of test organisms 73
3.11.3 Inoculation of test organisms 73
3.11.4 Test for anti-microbial activity 73
3.11.5 Determination of minimum inhibitory concentration 74
3.12 Procedure for Fourier Transform Infrared (FTIR) 75
3.13 Docking Procedure and Analysis 75
3.14 Statistical Analysis 76
CHAPTER FOUR: RESULTS AND DISCUSSION 77
4.1 Results 77
4.1.1 Qualitative phytochemical analysis of gouania longipetala 77
4.1.2 Quantitative phytochemical analysis of gouania longipetala
leaf extract 79
4.1.3 Acute toxicity evaluation of Gouania longipetala leaf extract 81
4.1.4 Antimicrobial activities of Gouania longipetala 84
4.1.5 Castor-oil induced diarrhea 87
4.1.6 Castor oil- induced enteropooling 91
4.1.7 Assay of fractions on castor oil- induced diarrhea 93
4.1.8 Assay of fractions on castor oil- induced enteropooling 97
4.1.9 Measurement of charcoal meal transit time 101
4.1.10 Evaluation of fractions of the extract on charcoal meal transit time 104
4.1.11 Estimation of serum electrolytes for castor oil-induced antidiarrhoeal
Model 107
4.1.12 Evaluation of effects of fractions and crude extract on electrolyte levels 109
4.1.13 Result of GC-MS analysis of the crude extract 111
4.1.14 Result of molecular docking of Gouania longipetala leaf extract 116
4.2 Discussion 122
CHAPTER FIVE: SUMMARY AND CONCLUSION 131
References 133
LIST OF TABLES
3.1: Solvent proportion 69
3.2 Pooling of fractions 72
4.1: Qualitative phytochemical composition of Gouania longipetala leaf
extract 78
4.2: Quantitaive phytochemical composition of G. longipetala leaf extract 80
4.2 Result of acute toxicity evaluation of Gouania longipetala leaf extract 80
4.3: Phase 1 result of acute toxicity evaluation of Gouania longipetala leaf
extract 82
4.4: Phase 2 result of acute toxicity evaluation of Gouania longipetala
leaf extract 83
4.5: Diameter of zones of inhibition of the ethanol extract of
gouania longipetala on the test bacteria isolates (mm) 86
4.6: Effect of the ethanol extract of gouania longipetala on castor
oil-induced diarrhea 89
4.5 Castor oil- induced enteropooling 90
4.6 Assay of fractions on castor oil- induced diarrhea 92
4.7: Effect of ethanol extract of leaves of Gouania longipetala on castor
oil-induced enteropooling in experimental albino rats 92
4.8: Effect of six different fractions and the crude extract of
Gouania longipetala leaves on castor oil- induced diarrhea in
experimental rats 94
4.9: Effects of six different fractions and crude extract of Gouania longipetala
on castor oil induced enteropooling in experimental albino rats 99
4.10: Effect of the ethanol extract of the leaves of Gouania longipetala on
charcoal meal test intestinal transit time 103
4.11: Effect of six different fractions and crude extract of leaves of
Gouania longipetala on charcoal meal intestinal transit time 106
4.12: Effects of the ethanol extract of the leaves of Gouania longipetala
on concentration of electrolytes in the intestinal contents 108
4.13: Effect of six different fractions and crude extract of the leaves of
Gouania longipetala on concentration of electrolytes in the
intestinal contents 110
4.14: GC-MS spectral analysis of ethanolic extract of leaves of
Gouania longipetala 112
4.15: Binding affinities of the best ligand poses with specific macromolecules 118
LIST OF FIGURES
4.1: Inhibition of stooling frequency in rats following treatment with
graded doses of ethanol extract of Gouania longipetala and also
the standard drug (Loperamide). 90
4.2: Percentage inhibition in stooling frequency following treatment with
the the crude extract and fractions of Gouania longipetala leaves
extract and also the standard drug (Loperamide). 96
4.3: Inhibition of enteropooling 100
4.4: 2D molecular interactions of best docking models of ligands vs macromolecule(6PT3). 119
LIST OF PLATES
2.1: Gouania longipetala plant photographed at a bush in Nsukka,
Enugu State, Nigeria 43
2.2: Gouania longipetala plant photographed from a bush in Nsukka,
Enugu State, Nigeria 43
LIST OF ABBREVIATIONS
AAP - American Academy of Pediatrics
AMPs – Antimicrobial proteins
CD – Celiac disease
CNS – Central nervous system
COX – Cyclooxygenase
CRC - Colorectal cancer
DPP-IV – Dipeptidylpeptidase
DRA - Down Regulated in Adenoma
EC- Epithelial cells
EEC – Enteroendocrine cells
ETEC – Enterotoxigenic E. coli
EIEC - Enteroinvasive E. coli
FTIR – Fourier transform infrared spectroscopy
GALT - gut-associated lymphoid tissue
GCMS – Gas chromatography and mass spectroscopy
GERD - gastroesophageal reflux disease
GLP 1 and 2 – Glucagon-like-peptide 2
GIP - G-insulinotropic polypeptide
HPA - hypothalamic pituitary axis
HIC - High-income countries
HIV – Human immunodeficiency virus
IBD – Inflammatory bowel disease
IBS – Inflammatory bowel syndrome
IECs – Intestinal epithelial cells
IL- Interleukin
JAMs - Junctional adhesion molecules
JAK - Janus kinase
LABs – Lactic acid bacteria
LMIC - low- and middle-income countries.
LPS – Lipopolysaccharides
mRNA – Messenger-RIBONUCLEIC ACID
TGFβ1 – Transforming growth factor beta1
ORT – Oral rehydration therapy
PCR - Polymerase chain reaction
PAT - Putative Anion Transporters
TJ – Tight junctions
T2D – Type 2 Diabetes
TLR4 – Toll-like receptor 4
TNFα – Tumour necrotic factor alpha
UC – Ulcerative colitis
WHO – World Health Organization
Reviews
No reviews yet.
APA
NWACHUKWU, & CHIWUBA, K. (2023). Anti-Diarrhoeal Potential Of Ethanol Extract of Gouania longipetala Leaves. Michael Okpara University of Agriculture. Retrieved June 7, 2026, from http://repository.mouau.edu.ng/works/anti-diarrhoeal-potential-of-ethanol-extract-of-gouania-longipetala-leaves-7-2
MLA
NWACHUKWU, and KENNETH CHIWUBA. "Anti-Diarrhoeal Potential Of Ethanol Extract of Gouania longipetala Leaves." Michael Okpara University of Agriculture, 15 Aug. 2023, http://repository.mouau.edu.ng/works/anti-diarrhoeal-potential-of-ethanol-extract-of-gouania-longipetala-leaves-7-2. Accessed June 7, 2026.
Chicago
NWACHUKWU, and KENNETH CHIWUBA. "Anti-Diarrhoeal Potential Of Ethanol Extract of Gouania longipetala Leaves." Michael Okpara University of Agriculture (2023). Accessed June 7, 2026. http://repository.mouau.edu.ng/works/anti-diarrhoeal-potential-of-ethanol-extract-of-gouania-longipetala-leaves-7-2