PHYTOCHEMICAL EVALUATION OF PLEUROTUS OSTREATUS GROWN ON HCL TREATED PALM OIL BUNCH

ABSTRACT

Plant foods are receiving more scientific attention because of their potentiall to ameliorate certain ailments in the human system. P.ostreatus is a fungi popularly called oyster mushroom, which has been known for producing biologically active secondary metabolites. This study was conducted with the objective to determine the concentrations of phytochemicals present in this edible mushroom grown on palm bunches soaked in varying concentrations of HCl ranging from 0.1M HCl to 0.5M HCl and control which was totally void of HCl. The phytochemical screening was carried out using standard methods. Results of this screening revealed the presence of phenols, flavonoids, alkaloids, terpenoids, tannins, glycosides, reducing sugars and steroids.  The result revealed that phenols, flavonoids, alkaloids and reducing sugars were present in appreciable quantities, while the others were low. Phenols was highest in 0.1M HCl with a mean±S.E.M of 188.96±6.01 mg/g while the lowest was found in 0.2M HCl with a concentration of 106.56±1.76 mg/g. Tannin was highest in 0.1M HCl, having a concentration of 5.89±0.83 mg/g and the least concentration was found in 5.24±0.04 mg/g. The HCl concentration with the highest flavonoid was 0.2M HCl (160.20±1.85 mg/g) while the least was in control (51.93±0.67 mg/g). Steroids was highest in control (0.48±0.01 mg/g) and least in 0.5MHCl (0.05±0.01 mg/g). Alkaloids was highest in 0.2M HCL (46.88±0.43) while the least was found in 0.5M HCl (24.10±0.94 mg/g). Terpenoids had the highest concentration in control (9.37±0.39 mg/g) while the least was found in 0.5M HCl (3.75±0.25 mg/g). Glycosides had the highest concentration in 0.2M HCl (1.98±0.00 mg/g) and the least in 0.5M HCl (1.67±0.00 mg/g). Reducing sugars was highest in control (23.95±0.96 mg/g) while the least was in 0.2M HCl (18.73±1.62). These phytochemicals have a broad spectrum of biological activities which includes antioxidant, anti-inflammatory, antiviral, antimicrobial, analgesic, immunostimulatory, hepatoprotective, antibacterial, antitumor anticancer amongst a host of others. This study therefore suggests that the cultivation and consumption of P.ostreatus on palm bunches soaked in varying HCl concentrations can have a positive effect on the health of the human body in several ways and can be harnessed in the management and control of diseases.

TABLE OF CONTENTS

Title page                                                                                                    i

Declaration                                                                                                 iii

Certification                                                                                                         iv

Dedication                                                                                                  v

Acknowledgement                                                                                                vi

Table of contents                                                                                        vii

List of tables                                                                                               xi

List of figures                                                                                             xii

Abstract                                                                                                      xiii

CHAPTER ONE

1.0 INTRODUCTION

1.1 Background of the study                                                                      1

1.2 Statement of the problem                                                                     2

1.3 Justification of the study                                                                      3

1.4 Objectives of the study                                                                         3

CHAPTER TWO

2.0 LITERATURE REVIEW

2.1 Edible mushrooms (pleurotus ostreatus)                                                         4

2.2    Mushrooms as therapeutic agents.                                                    6

2.2.1 Antioxidant activity                                                                                    7

2.2.2 Anti-inflammatory activity                                                                7       

2.2.3 Antitumor activity                                                                                      8

2.2.4 Immunomodulatory activity                                                              8

2.2.5 Antidiabetic effects                                                                                      8

2.2.6 Antiviral                                                                                            9       

2.3   Nutraceutical properties of pleurotus ostreatus                                           9

2.4   Phytochemicals                                                                                   10

2.5   Biological activities of phytochemicals                                                        11

2.5.1 Phenolic compounds                                                                         12

2.5.2 Flavonoids                                                                                         13

2.5.3 Tannin                                                                                               14

2.5.4 Alkaloids                                                                                           16

2.5.5 Saponins                                                                                           18

2.5.6 Cyanogenic glycosides                                                                       19

2.5.7 Terpenoids                                                                                        20

2.5.8 Steroids                                                                                             21

2.5.9 Reducing sugars                                                                                22

CHAPTER 3

MATERIALS AND METHODS

3.1.0 Source of raw materials                                                                     23

3.1.1 Spawn preparation                                                                            23     

3.1.2 Substrate preparation                                                                        24

3.1.3 Substrate inoculation                                                                         24

3.2    Equipment and apparatus                                                                           28

3.3:   Chemicals and reagents                                                                     29

3.4   Quantitative determination of phytochemicals in

         P.ostreatus mushroom.                                                                      29

3.4.1 Alkaloid determination                                                                      29

3.4.2 Flavonoids determination                                                                           30

3.4.3 Determination of steroids                                                                            30

3.4.4 Determination of terpenoid                                                               30

3.4.5 Determination of tannins                                                                            31

3.4.6 Determination of glycosides                                                              31

3.4.7 Determination of reducing sugars                                                      31

3.4.8 Determination of phenols                                                                            31

CHAPTER FOUR               

4.0 RESULT AND DISCUSSION                                                            34                           

4.1 Discussion                                                                                            35

CHAPTER FIVE                  

5.0 CONCLUSION AND RECOMMENDATION                                          

5.1 Conclusion                                                                                           38

5.2 Recommendation                                                                                  38

REFERENCES

APPENDIX

LIST OF TABLES

Table 3:1 equipment/ apparatus used and their model.                              28

Table 4.1: phytochemical result of mushroom grown on

                substrates of varying HCl

                concentrations                                                                                      34

LIST OF FIGURES

Fig.2.1: P.ostreatus grown at Michael Okpara University

  of agriculture, umudike         .                                                                  5

Fig. 2.2: structure of different flavonoids.                                                  13

Fig. 2.3: structure of typical tannins                                                           15

Fig. 2.4: structure of different classes of alkaloids.                                          17         

Fig. 2.5: structure of typical glycosides.                                                     19

Fig. 2.6: structure of monoterpenoid and sesquiterpenoid                                   21

Fig. 2.7: structure of a typical phytosterol                                                 22

Fig. 3.1: Flow chart for spawn production.                                                         26

Fig. 3.2 Flow chart for substrate preparation.                                            27     

CHAPTER ONE

1.0 Introduction

1.1 Background of the study

 The word "Phytochemicals" was derived from the Greek word "phyto", meaning plants. Plants are made up of chemicals of various kinds (Breslin, 2017). These chemicals are known as phytochemicals. By definition, phytochemicals are defined as those chemicals that are produced by plants either through primary or secondary metabolism. Thus, they are referred to as secondary metabolites (Molyneux et al., 2007).

Phytochemicals can also be defined as biologically active, naturally occurring chemical compounds found in plants which protect plant cells from environmental hazards such as pollution, stress, drought, ultra violet exposure and pathogenic attack (Ali et al., 2006). Not only do they protect plants against environmental hazards, they also give protection against predators, thus they are said to serve protective function. They are not required for sustenance of life but possess important properties which help prevent or fight against some common diseases (Holst et al., 2008). Some phytochemicals serve antioxidant purposes which confers a medicinal property to the plants. Antioxidants are chemicals which scavenge free radicals which are produced as a result of some reactions taking place during cellular metabolism

The benefits of phytochemicals are numerous, they include low toxicity, low cost, easy availability, not excluding their biological benefits such as antimicrobial effects, stimulations of the immune system, decrease of platelet aggregation and modulation of hormone metabolism and anti-neoplastic properties. (Andre et al; 2010). Phytochemicals include flavonoids, alkaloids, glycosides, oxalates, phytates, tannins, saponin etc.

Pleurotus ostreatus is an edible mushroom with fleshy spores bearing fruiting body of a fungus, typically produced above or beneath the ground which have been found to be growing on decaying trees and can be harvested wild or cultivated. (Valverde et al., 2015).

It has been proven that some species of mushrooms have outstanding and remarkable bioactivities such as antioxidant, antitumor, antiviral, anti-inflammatory, anticomplementary, anticoagulant, antidiabetic, hypolipidemic, hepatoprotective, immunostimulant and immunological activities which made them suitable for use in food, cosmetics and biomedicine (Loria et al., 2014).

The phytochemicals present in Pleurotus ostreatus confers an antioxidant property to it and as such is highly recommended as a natural source of antioxidant since synthetic antioxidants have been found to produce side effects dangerous to the health (Kozarski et al., 2014). Mushrooms aid in enhancing antioxidant defense through diets to reduce the level of oxidative stress caused by reactive oxygen species (ROS).

1.2 STATEMENT OF THE PROBLEM

The nutritional value of P.ostreatus is well known but the phytochemical constituent needs to be emphasized to provide knowledge of their biological functions when consumed. The phytochemical evaluation of P.ostreatus goes beyond its nutritional value.

1.3 JUSTIFICATION OF THE STUDY

The younger generation who are now used to meat and fish are ignorant of the use of edible mushrooms which they find growing around them. The knowledge of the phytochemical properties of P.ostreatus will lead to a better understanding of its nutritive and medicinal role in human health and can as well lead to an increase in the cultivation and consumption of this mushroom.

1.4 OBJECTIVES OF THE STUDY

The main objective of this research work is to determine the phytochemicals present in P.ostreatus. grown on palm bunches soaked in HCl concentrations ranging from 0.1M HCl to 0.5M HCl and control which was totally void of HCl.

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