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Antioxidant potentials of the methanol leaf extract of Piper guineense (Uzuza) leaves was evaluated using standard analytical procedures. Properties such as phytochemicals, proximate composition, mineral composition and total antioxidant potentials were determined. The result of the phytochemical analysis showed the presence of alkaloid (20.67 ± 0.85 %), flavonoid (0.70 ± 0.02 %), phenol, (7.21 ± 0.21 %), tannins (0.72 ± 0.01 %), saponins (14.82 ± 0.36 %) and HCN (126.00 ± 10.58 mg/kg). The proximate composition assay revealed that the plant contains moisture content (11.40 ± 0.37 %), crude fat (1.90.00 ± 0.07 %), ash content (17.30 ± 0.46 %), crude fibre (21.64 ± 0.80 %), crude protein (13.62 ± 0.57 %) and carbohydrate (45.54 ± 1.00 %). The mineral elements present were copper (75.1 ± 0.44 mg/kg), iron (33.80 ± 0.53 mg/kg), magnesium (23.60 ± 0.60 mg/kg), manganese (74.00 ± 1.00 mg/kg) and zinc (124.00 ± 10.21 mg/kg). The total antioxidant potentials of Piper guineense leaf extract was 16.5mg equivalent. A graph change of change in absorbance against concentration of the standard (ascorbic acid) (mg/L) and the graph of change in absorbance against the concentration of the extract (mg/L) was plotted. The result showed that piper guineense  leaf aside from having valid nutritional and therapeutic potentials, is also a good source of antioxidant that is capable of enhancing human nutraceutical needs.


Cover page                                                                                                                              i

Title page                                                                                                                                ii

Certification                                                                                                                            iii

Declaration                                                                                                                              iv

Dedication                                                                                                                              v

Acknowledgement                                                                                                                  vi

Table of Contents                                                                                                                   vii

List of Tables                                                                                                                          viii

List of Figures                                                                                                                         ix

List of Plates                                                                                                                           x

Abstract                                                                                                                                  xi        


1.1       Background to the Study                                                                                           1

1.2       Aims and Objective of the Study                                                                               4

1.3       Statement of Problem                                                                                                 5

1.4       Justification of the Study                                                                                           5

1.5       Scope and Lamination of the Study                                                                           6


2.1        Morphology and Description of Piper guineense                                                      7

2.2        Phytochemicals lsolated from Piper guineense                                                         8

2.3       Nutritional value                                                                                                         12

2.4       Uses of Piper guineense                                                                                             13

2.5       Antioxidants                                                                                                               15

2.5.1    Natural and synthetic antioxidants                                                                             17

2.6       Phytochemicals                                                                                                           22

2.6.1    Alkaloids                                                                                                                     22

2.6.2    Saponin                                                                                                                       23

2.6.3    Phenols                                                                                                                        24

2.6.4    Tannins                                                                                                                        25

2.6.5    Flavonoids                                                                                                                  25

2.6.6    Hydrogen cyanide (HCN)                                                                                          26

2.6.7     Cardiac glycosides                                                                                                     27

2.7       Proximate Composition                                                                                              27

2.7.1    Moisture content                                                                                                         28

2.7.2    Ash residue                                                                                                                 28

2.7.3    Total carbohydrates                                                                                                    29

2.7.4    Crude fat                                                                                                                     29

2.7.5    Crude protein                                                                                                             30

2.7.6    Crude fibre                                                                                                                  30

2.8       Mineral Elements                                                                                                        31

2.8.1    Copper (Cu)                                                                                                                32

2.8.2    Iron (Fe)                                                                                                                      32

2.8.3    Manganese (Mn)                                                                                                         33

2.8.4    Zinc (Zn)                                                                                                                     33

2.8.5    Magnesium (Mg)                                                                                                         33


3.1       Materials Used                                                                                                            34

3.2        Plant materials                                                                                                            34

3.3       Sample Preparation                                                                                                     34

3.4        Extraction                                                                                                                  34

3.5       Phytochemical Analysis                                                                                              35

3.5.1    Determination of saponins                                                                                          35

3.5.2    Determination of tannins                                                                                            35

3.5.3    Determination of alkaloids                                                                                         36

3.5.4    Determination of phenol                                                                                             37

3.5.5    Determination of flavonoids                                                                                       37

3.5.6    Determination of cyanogenic glycoside (HCN)                                                         38

3.6       Proximate Composition                                                                                              38

3.6.1    Determination of ash residue                                                                                      38

3.6.2    Moisture of content determination                                                                             39

3.6.3    Fat determination                                                                                                        39

3.6.4    Crude fibre determination                                                                                          40

3.6.5    Protein determination                                                                                                 40

3.6.6    Determination of carbohydrate                                                                                   42

3.7       Determination of Minerals                                                                                          42

3.7.1    Preparation of ferric reducing antioxidant power (FRAP) reagent                            42

3.7.2    Serial dilution of the stock solution of ascorbic acid                                                  43

3.7.3    Measurement of total antioxidant potentials of the methanol exract

using FRAP method                                                                                                   44


3.8       Statistical Analysis                                                                                                      44


4.1       Phytochemical                                                                                                             46

4.2       Proximate Composition                                                                                              47

4.3       Mineral Element                                                                                                          49

4.4       Result of FRAP Assay                                                                                               50                   


5.1       Conclusion                                                                                                      54

5.2       Recommendation                                                                                            54









2.1       Ethanomedicinal uses of Piper guineense                                                                  15


4.1       The results of qualitative and quantitative phytochemical

analysis                                                                                                                        45


4.2       The results of proximate analysis                                                                                47


4.3       The results of qualitative and quantitative mineral element

Analysis                                                                                                                      49


4.4       Results of FRAP Assay                                                                                                          50


4.5       Concentration of Standard and change in absorbance                                               51


4.6       Concentration of Methanol extract and change in absorbance                                   52








2.1       Chemical structure of some Phytochemcials Isolated

from Piper guineense                                                                                      12


2.2       Different types of Antioxidant compounds (Rajani, 2004)                                       16


2.3       (a) Natural Antioxidant (Tocopherol), (b) Synthetic

antioxidant (BHA)                                                                                                      18


2.4        Antioxidant reaction                                                                                                  19


2.5       DPPH Radicals Scavenging Capacity: Transformation of

DPPH form to non-radical form                                                                                 20


2.6       Transformation of ABTS radical form to reduced

non-radicals form                                                                                                        21


2.7       FRAP antioxidant reaction with ferric salt                                                                 22


2.8       The structure of Alkaloid – Piperine                                                                           23

2.9       Structure of Saponin – Glycoside                                                                               24

2.10     Structure of phenol – flavone                                                                                                 24

2.11     Structure of Tannins Theaflavin                                                                                 25

2.12     Structure of Flavonoids - Flavan                                                                                26

2.13     Structure of Cyanide                                                                                                  27

2.14     Structure of Cardia glycosides                                                                                   27


4.1       Absorbance against concentration of the Standard (ascorbic acid)                            51


4.2       Absorbance against concentration of Standard (Mg/L) and

that of  the  extract.  (µg/L)                                                                                        52








2.1                   Piper guineense showing (A) Ripe seeds (B) Leaves

            (C) Unripe seeds                                                                                 8









An antioxidant is a chemical substance that inhibits the oxidation of other chemicals. They preserve the vital key cell constituents in the human body by counteracting the harmful effects of free radicals, which are natural byproducts of cell metabolism (Miller et al., 2000). The oxidative stress (OS) as influenced by reactive oxygen species (ROS) can be described as a dynamic imbalance between the amounts of free radicals generated in the body and levels of antioxidants to douse and/or scavenge them and safeguard the body against their harmful consequence (Shirwaikar et al., 2006).


The World Health Organization (WHO) estimated that 80% of the populations of developing countries in Asia and Africa rely on traditional medicines, mostly plant drugs, for their primary health care needs. (WHO, 2008).

Recently, there has been increase on the use of medicinal plants in the developed countries and this has been linked to the distillation and production of various medicines and chemotherapeutics from these plants as well as from locally used herb remedies. The medicinal properties of these plants could be hinged on the antioxidant, antimicrobial, antipyretic impact of the secondary metabolites present in them (Soetan and Iyelaagbe, 2009).

Rephrage, plant extracts and plant-derived drugs have contributed immensely to the overall health and wellbeing of man (Anyanwu and Nwosu, 2014).

In 2008, World Health Organization highlighted the significance of scientific research into alternative medicine. Herbs are widely evaluated for their attributes as food as well as medicine. Many under developed nations in the world rely on herbs as a possible addition to World Health Organization collection of “important drugs” once their benefit has been clinically proven.

 Of the 265,000 species of flowering plants that have been identified on planet earth, only 0.5% of them have been studied in detail for chemical structure and therapeutic value (Omodamiro and Ekeleme, 2013).


Infact, modern scientists only know the chemical composition of less than 5% of the flora in the rainforest (Jackson, 1989). At least 12,000 such compounds have been isolated and this number is estimated to be less than 10% of the total (Lai and Roy, 2014; Tapsel et al., 2006).

Chemical composites in plants regulate their outcome in the human body via procedures similar to the ones that are well known for the chemical compositions in orthodox drugs. Therefore, herbal medicines are not different from orthodox ones in terms of their curative effect. This makes herbal medicines to be as efficient as orthodox medicines but also present them with similar capacity to engender damaging consequence.

Research into uses of plants has been accepted as a result of efficient means to determine future medicines. The use of these plants in medicine is due to the availability of phytochemical compounds such as phenols, flavonoids, tannins and alkaloids, present either in the seeds, leaves, stems and roots (Tapsel et al., 2006).

Medicinal plants have been used since medieval times as origin of medicine for the treatment of all kinds of ailments. Traditional medicine and herbs remain functional in the primary healthcare systems of most cultures.

About 25% of the drugs prescribed worldwide and found in modern pharmacopoeias come from plants, 121 such active compounds being in current use. Of the 252 medicines regarded as fundamental and necessary by the World Health Organization (WHO), 11% are sourced from plant and a significant percentage are artificial medicines derived from natural sources. Examples of essential medicines derived from plants include; digoxin from Digitalis spp. used for managing congestive cardiac heart failure (CCF), quinine (antimalarial) and quinidine (antiarrythmic) from Cinchona spp., vincristrine and vinblastine (anticancer agents) from Catharanthus roseus, atropine from Atropa belladonna and morphine and codeine from Papaver somniferum. It is calculated that 60% of drugs use in treating tumour and infections are currently in the market or undergoing clinical trial are from plants. (Rates, 2001).

This resurgence of interest in medicinal plants and traditional medicine as a whole is owing to the fact that, conventional drugs could be ineffective (e.g. side effects and ineffective therapy), wrong dosage of chemically formulated medicines may cause harmful effects and related problems, a greater part of the global population do not use synthetic drugs for treatment, perhaps due to inaccessibility of healthcare centers and the ever increasing cost of synthetic drugs. Finally, traditional medicine and environmental consciousness imply that herbal derivatives are harmless. (Rates, 2001).

Medicinal plant extracts comprise of diverse bioactive substance with diversities of biological activities of valued beneficial guides. The protective effects of plant are attributed to the activities and potentials of bioactive substances (Etim et al., 2013), which are the non-nutrient compounds. They are classified into three main groups namely alkaloids, isoprenoids (terpenoids and steroids) and phenols (phenolic acid, flavonoids, tannins and coumarin). The plant derived phytoconstituents are capable of terminating free radical reactions and hinder the body or biological systems from oxidative damage and protect against chronic diseases (neurodegenerative and cardiovascular (Etim et al., 2013).

Oxidative stress is defined as a disruption in the balance between the making of reactive oxygen species (ROS), free radicals and antioxidant defenses (Eboh, 2014). This imbalance leads to damage of important molecules such as proteins, lipids, nucleic acids and is also involved in the pathogenesis of various forms of diseases including cardiovascular diseases; cancer etc. Moreover, antioxidants are different group of chemicals that inhibit the oxidation of other molecules, neutralizing free radicals and protect specific organs against deleterious effect of dangerous xenobiotics (Durackova, 2010; Shirwaikar et al., 2006).

The deleterious impacts of reactive oxygen species (ROS) are eradicated by antioxidants and are of two kinds, namely, enzymatic and non-enzymatic (Eboh, 2014).

Halliwell and Gutteridge, (1989); Huang et al., (2005), defined antioxidant in food science as a constituent in foods when present at low concentration compared to those of oxidizable substrate greatly reduces or inhibits negative impacts of reactive species, such as reactive oxygen and nitrogen species (ROS/RNS), on normal biological roles in humans. Ndiamaka, (2019), reported that, “antioxidants are substance that are able to prevent the oxidation of easily oxidization materials.”

Natural and artificial antioxidants are extensively applied in conventional medicine. Some of them have been proven to be effective protectors. That is, they prolong the life span of laboratory animals when applied to food or drinking water on a regular basis. In vitro, antioxidants restrain free radical chain oxidation reactions, culminating in oxidation of fatty acids, edible fats etc. More so, their effectiveness as scavengers of oxygen free radicals in cells and tissues is insignificant in relationship to natural antioxidant enzymes (Koltover, 2010).


            1.2              AIMS AND OBJECTIVE OF THE STUDY

This Project work aims to extract and isolate antioxidant principles from methanol leaf extract of Piper guineense (Uziza) using the following specific objectives:

i)                    To obtain the methanol leaf extract by cold maceration.

ii)                  To subject the methanol leaves extract to phytochemical screening (qualitative and quantitative), proximate and mineral element composition.

iii)                To determine the total antioxidant potentials in   Piper guineense leaves extract.

iv)                To draw conclusion and offer suggestions on the possible therapeutic utilization of the Piper guineenses leaves based on the result obtained.



While a lot has been reported about nutritional and health values of Piper guineense (Uziza) leaves, there is very little scientific evidence for verifying the efficacy of the antioxidant properties, possible health and nutritional values of Piper guineense   leaves. Lack of information on the economic values of Piper guineense   leaves resulted in the under cultivation and utilization of its products. Hence, this research aimed at evaluating the phytochemical, micro nutritional and antioxidant characteristics of Piper guineense leaf.


The paucity of knowledge of the phytochemical constituents, antioxidant and nutritional benefits of Piper guineense (Uziza) leaves has resulted in its neglect and under-utilization.  It is envisioned that the outcome of this research will motivate the extraction of the preservative, nutraceutical and healing capacities of these cooking herbs.  Nigeria is home to great biological diversity including indigenous plants from which potential lead structures for use in formulation of novel drugs can be obtained. Piper guineense is used in traditional medicine to boost uterine compression for the removal of placenta during child birth, regulation of weight and treatment of pains. Anti-parasitic, antimicrobial and antifungal activities of Piper guineense leaf has also been reported. Such knowledge will inform pharmaceutical industry on the therapeutic importance of these plants, and will also validate indigenous knowledge as valuable resource for medicine and pharmaceutical explorations.


The experiments will focus mainly on determining the phytochemical constituents, antioxidant and nutritional benefits of Piper guineense (Uziza) leaves. The study covers nutritional benefits and antioxidant benefits in order to establish the indigenous claim that Uziza has some medicinal and pharmaceutical potentials.


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