EVALUATION OF PHYTOCHEMICAL, ANTI-INFLAMMATORY AND DIURETIC ACTIVITIES OF ETHANOL LEAF EXTRACTS OF PTERIDIUM AQUILINUM (L.) KUHN AND SCLERIA VERUCOSA WILLD.

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ABSTRACT

 

This work investigates the phytochemical analysis, diuretic and anti-inflammatory properties of ethanol leaf extract of P. aquilinum and S. verucosa. Wistar rat model was used in testing the anti-inflammatory and diuretic activities of the two plants. The leaves of the plants were collected, sun-dried, milled and stored for analysis. Some of the milled samples were soaked in ethanol for 48 hours after which it was filtered and the filtrate was subsequently evaporated to dryness and stored for further use. A total of 56 Wistar rats were used for this study. They were grouped into two major groups: diuretic and anti-inflammatory groups. Phytochemical screening (quantitative and qualitative), proximate and mineral compositions screening were done using standard procedures of analysis. Phytochemical results show that the plant, P. aquilinum has significant (p<0.05) concentrations of total phenolics (66628.50 ± 564.79), carbohydrates (4108.24 ± 211.44) and reducing sugars (753.29 ± 22.27) when compared with S. verucosa viz: 22806.45 ± 237.49, 1302.11 ± 23.18 and 596.54 ± 13.63, respectively, while S. verucosa showed significant (p<0.05) concentrations of terpenoids (967.23 ± 9.85), steroids (64.21 ± 15.95) and flavonoids (3058.13 ± 29.47) when compared with P. aquilinum viz: 749.91 ± 6.08, 19.12 ± 0.21 and 944.38 ± 2.25, respectively. Also, anti-inflammatory study indicated that the result of C-reactive protein had a non-significant (p>0.05) and significant (p<0.05) dose dependent decrease when the groups administered low (2.66 ± 0.18) and high (2.03 ± 0.26) doses, respectively of S. verucosa are compared with the inflammation (no treatment) group (2.58 ± 0.03). The groups administered low (2.54 ± 0.07) and high (2.15 ± 0.06) doses of P. aquilinum show significant (p<0.05) decrease when compared with the no treatment group. The result of Nitric oxide shows that there is a non-significant (p>0.05) difference when the treated groups are compared with the normal control as well as the non-treated group, while the group treated with low dose of S. verucosa (25.14 ± 0.33) shows a significant (p<0.05) increase when compared with all the groups, but non-treated group. Lactate dehydrogenase result shows that there is a non-significant (p>0.05) difference when the groups treated with ethanol extracts of the plants are compared with the normal control and other control groups. In diuretic activity, results of sodium, chloride and potassium in both urine and faeces samples indicate that the test groups administered the different extracts showed a significant decrease when compared with the positive control, the bicarbonate concentration in both specimens was of no significant difference when compared with the positive control. Therefore, this thesis indicates that the plants, P. aquilinum and S. verucosa have some medicinal and nutritional benefits.









TABLE OF CONTENTS

 

 

Title Page                                                                                                                                ii

Declaration                                                                                                                             iii

Certification                                                                                                                            iv

Acknowledgements                                                                                                                v

Table of Contents                                                                                                                   vi

List of Tables                                                                                                                          viii

List of Figures                                                                                                                         ix

List of Plates                                                                                                                           x

Abstract                                                                                                                                  xi

 

CHAPTER 1

INTRODUCTION

 

1.1        Background of the Study                                                                                      1

1.2       Statement of Problem                                                                                                 2

1.3       Justification of the Study                                                                                            3

1.4       Aim of Study                                                                                                              4

1.5       Objective of Study                                                                                                      4

 

CHAPTER 2

LITERATURE REVIEW

 

2.1       The Phyto-Medicinal Chemistry of Plants                                                                 5

2.2       Mineral Composition of Plants                                                                                   11

2.3       Proximate Composition of Plants                                                                               15

2.4       Anti-Inflammatory Activities of Medicinal Plants                                                     15

2.5       Diuretic Activity of Plants                                                                                          19

 

CHAPTER 3                                 

MATERIALS AND METHODS

 

3.1       Collection and Identification of Plant Materials                                                  22

3.2       Processing                                                                                                            24

3.3    Preparation of Extracts                                                                                                  25

3.4    Phytochemical Screening of Extracts                                                                            25

3.4.1    Qualitative phytochemical Analysis of Extracts                                                        25

3.4.2    Quantitative Phytochemical Analysis of Extracts                                                      28

3.5       Proximate Analysis of the Plants                                                                                31

3.6       Mineral Analysis of P. aquilinum and S. verucosa                                                    36

3.7       Animal Source and Grouping                                                                                     38

3.8       Sample Collection                                                                                                      40

3.9       Anti-Inflammatory Test                                                                                              41

3.10     Diuretic Test                                                                                                               42

3.11     Statistical Analysis                                                                                                     44

 

CHAPTER 4

RESULT AND DISCUSSION

 

4.1       Results                                                                                                                        44

4.2       Discussions                                                                                                                 58

 

CHAPTER 5

CONCLUSION AND RECOMMENDATIONS

 

5.1       Conclusion                                                                                                                  65

5.2       Recommendations                                                                                                      65

References                                                                                                                  66

Appendix                                                                                                                    85

           

 

 

 


 

LIST OF TABLES

Table 4.1        Qualitative analysis of ethanol leaf extracts of P. aquilinum and

S. verucosa                                                                                                      44

Table 4.2        Quantitative analysis of ethanol leaf extracts of P. aquilinum and

S. verucosa                                                                                                      45

Table 4.3        Proximate composition of ethanol extracts of P. aquilinum and

S. verucosa                                                                                                      46

Table 4.4        Mineral composition of ethanol extracts of P. aquilinum and S. verucosa           47

Table 4.5        Anti-inflammatory activity of P. aquilinum and S. verucosa                                  49

Table 4.6        Diuretic activity of P. aquilinum and S. verucosa                                          53

 

 

 

 

 

LIST OF FIGURES

 

Fig. 4.1:          Effects of ethanol Leaf extracts of P. aquilinum and S. verucosa on

Serum C - reactive protein concentration in Wistar rats.                             49

Fig4.2:            Effects of ethanol extracts of P. aquilinum S. verucosa and on

Serum Nitric Oxide concentration in Wistar rats.                                          50

Fig. 4.3:          Effects of ethanol extracts of P. aquilinum and S. verucosa on

Serum lactate dehydrogenase activity in Wistar rats.                                     51

Fig. 4.4:          Effects of ethanol extracts of P. aquilinum and S. verucosa on

Serum Sodium ion concentration in Wistar rats.                                            54

Fig. 4.5:          Effects of ethanol extracts of P. aquilinum and S. verucosa on

Serum Chloride ion concentration in Wistar rats                                           55

Fig. 4.6:          Effects of ethanol extracts of P. aquilinum and S. verucosa on

Serum Bicarbonate ion concentration in Wistar rats                                      56

Fig. 4.7            Effects of ethanol extracts of P. aquilinum and S. verucosa on

Serum  Potassium ion concentration in Wistar rats                                        57

 

 

 

 

 

 

 

LIST OF PLATES

 

Plate 1:           Freshly collected leaves of Pteridium aquilinum                                           22

Plate 2:           Freshly collected leaves of Scleria verucosa                                                 23

Plate 3a:         Blended leaves of Pteridium aquilinum                                                         24

Plate 3b:         Blended leaves of Scleria verucosa                                                                24

Plate 4a:         Sacrificing the Wistar Rats for Sample collection                                         40

Plate 4b:         Collection of Samples from the sacrificed Wistar Rats                                    40


 

                                                                     

                                      

 

 

 

 

 CHAPTER 1

INTRODUCTION

1.1        BACKGROUND OF THE STUDY

The high usage of plant extracts in pharmaceutical industries makes the study of medicinal plants very important (Nostro et al., 2000). The acceptance of herbal medicine as an alternative means of healthcare and the incessant failure of treatment gave room for medicinal plant research (Hammer et al., 1999).

Pteridium aquilinum (Bracken Fern) is a plant under the phylum pteridophyta. Ferns and their likes consist of about 250 genera and 12000 species. The plants possess xylem and phloem which make them vascular plants. They have stems and roots like other vascularized plants (Chapman, 2010; John et al., 2012). P. aquilinum is commonly called bracken. It is a fern-like herb which belongs to the family Pteridaceae. It is a vascular plant with alternating generations of both sporophyte and gametophyte. It has bipinnate leaf structure. P. aquilinum has a cosmopolitan distribution, occurring in both wet and dry places. (Clapham et al., 1968)

Ferns possess different kinds of phytochemicals which can be beneficial to the health of mankind. Some of these phytochemicals act as antioxidants, antitoxins, anti-inflammatory and diuretics (Hassan et al., 2007; Awe and Amobi, 2015).

In the recent times, the bioactivities of ferns were analysed and they were reported to possess antioxidant and antimicrobial activities. The antioxidant activity is the most essential for human health (Chen et al., 2007; Ding et al., 2008). Scleriaverucosa commonly known as sedge is a plant which belongs to the family Cyperaceae. It is a monocotyledonous flowering plant which specifically resembles grasses. The leaves are greenish and bladed (Burkil, 1985).

1.2       STATEMENT OF PROBLEM

Since many years of existence of the mankind, many diseases abound. Most of these diseases are caused by the virulence of pathogenic micro-organisms while some arise due to physiological and environmental factors. As many diseases of human abound in the universe, so do green plants which undoubtedly possess the necessary phytochemicals that can fight these diseases of mankind.

However, the failure of man to identify these medicinal plants and their associated phytochemical constituents has continued to fester the virulence of these diseases on the human race. These varying diseases of human constitute serious issues and challenges to our helpless society and so need to be confronted via medicinal plants researches geared towards development of new drugs. Today, it is pathetic to behold the high prevalence of many health challenges that were non-issues during the olden days. These diseases have become rampant in our present society.

Lower plants like Bracken Fern (Pteridium aquilium) and Sedge (Scleria verucosa) abound in our environment. Despite the abundance of the lower plants enough researches have not been carried out to investigate their medical potentials like the higher plants. This challenge gave room for this research work. In a bid to find a solution to the dangers posed by many recurring diseases of man, Bracken ferns (P. aquilinum) and Sedge (S. verucosa) were assayed to determine their phytochemicals, proximate and minerals potentials as well as their diuretic and anti-inflammatory activities.

1.3       JUSTIFICATION OF THE STUDY

Plants with alkaloids, tannins, flavonoids, saponins, glycosides and phenolics are known to possess antimicrobial, antioxidant and diuretic activities (Hostettmann et al., 1995; Isaac and Chinwe, 2001). Since the existence of these active compounds in plants constitutes well-pronounced health benefits, the two plants (Pteridium aquilium and Scleria verucosa) were analysed for the existence of these compounds. The medicinal efficacies of these active compounds were tested using animal model. This research is meant to assist in the discovery of medicines for the cure of various diseases of man via medicinal plants research.

Many herbal medicine practitioners extensively use Pteridium aquilinum and Scleria verucosain preparing many combination therapies for ailing persons. So, the curiosity over the medicinal potentials of these plants gave birth to this reseach work. This investigative researchwas carried out with a view to revealing the bioactive compounds occurring in the twoplants and the therapeutic effects of the compounds via in-vivo studies.

The two African plants, Pteridium aquilinum and Scleria verucosa were used in this medicinal plant research. Phytochemical analysis was conducted on the ethanol leaf extracts of the two plants to determine their various phytochemical potentials. Also, proximate and mineral analyses were carried out to ascertain their mineral and proximate compositions. In other parameters, the anti-inflammatory and diuretic activities of the ethanol leaf extract of P. aquilinum and S. verucosa were tested using Wistar rats.

Phytochemical analyses of Pteridium aquilinum and Scleria verucosa serve to test, verify or falsify previous medicinal plant researches. This research serves to open a door to further research that will be harnessed by phytochemists and pharmacists in the development of new drugs that can withstand the drug resistance posed by many diseases.

1.4       AIM OF STUDY

To evaluate the phytochemical, anti-inflammatory, and diuretic activities of the ethanol leaf extracts of Pteridium aquilinum (L.) Kuhn and Scleria verucosa Willd.

1.5       OBJECTIVE OF STUDY

The objectives of this study are to:

i.             Determine the qualitative phytochemical properties of ethanol extract of Pteridium aquilinum (L.) Kuhn and Scleria verucosa Willd.

ii.            Determine the quantitative phytochemical properties of ethanol extract of Pteridium aquilinum (L.) Kuhn and Scleria verucosa Willd.

iii.          Determine the proximate composition of ethanol extract of Pteridium aquilinum (L.) Kuhn and Scleria verucosa Willd

iv.          Determine the mineral composition of ethanol extract of Pteridium aquilinum (L.) Kuhn and Scleria verucosa Willd.

v.            Determine theanti-inflammatory activities of ethanol leaf extracts of Pteridium aquilinum (L.) Kuhn and Scleria verucosa Willd in Wistar rats.

vi.          Determine the diuretic activities of ethanol leaf extract of Pteridium aquilinum (L.) Kuhn and Scleria verucosa Willd in Wistar rats.


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