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The bioactive ingredients of three Nigerian spices: Cocoplum (Chysobalanus icaco), African black pepper (Piper guineense) and Aidan fruit (Tetrapleura tetraptera) blends were comparatively assessed in this study. The spices were separately washed with cold water, air dried at room temperature (27±2)oC ,milled, sieved (2mm) and formulated into 9 samples :20:15:65, 56:24:20, 75:15:10, 0:0:100, 47:42:11, 100:0:0, 47:15:38, 29:25:46 and 0:100:0% Cocoplum, African black pepper and Aidan fruit spices) as derived from mixture design of Design Expert-6.0.8 Software. The spices were subjected to proximate, mineral, vitamin, antioxidant and bioactive analysis in duplicates using standard analytical methods. Analysis of variance was carried out on the data obtained using Statistical Package of Social Science version 17.0. Result of proximate analysis revealed the moisture content(8.83 to 14.62%),crude protein(5.41 to 10.92%),crude fibre (0.95 to 8.5%),fat (6.37 to11.55%),ash(3.84to10.34%)and carbohydrate(52.62 to 71.45%)content of the spices. The mineral content of the spices showed that phosphorus ranged from 14.63 to 87.93mg/100g,calcium ranged from 19.77 to54.04mg/100g,magnesium ranged from 15.03 to 34.24mg/100g,sodium ranged from 5.86 to18.77mg/100g,whereas iron, manganese and potassium within 1.27 to 3.51mg/100g,4.38 to 21.68 mg/100g manganese, and 10.44 to54.63 mg/100g respectively. Result of vitamin content revealed the pro-vitamin A (1.56 to 7.07 µg/100g), thiamin(0.25 to9.04 mg/100g),riboflavin(0.38 to 4.52 mg/100g), ascorbic acid (0.74 to16.39 mg/100g).The bioactive compounds of the spices revealed that tannin ranged from 0.37 to1.74 mg/100g,flavonoid ranged from 2.06 to 5.82 mg/100g,alkaloid ranged from 1.63 to 4.06 mg/100g,phenol ranged from 0.33 to 1.05 mg/100g,saponin ranged from 0.65 to 2.02 mg/100g, steroid ranged from 0.13 to 0.40 mg/100g,terpenoid ranged from 0.12 to 0.38 mg/100g,whereas trypsin inhibitor ranged from 0.15 to 0.43 mg/100g.Result of the antioxidant properties showed that FRAP ranged from 10.07 to 20.77 mg/100g whereas DPPH ranged within10.52 to 40.28%.It can be deduced from this study that nutritious spices can be processed from blends of Indigenous and underutilized crops like Cocoplum, African black pepper and Aidan fruit.



Cover Page                                                                                                                              i

Title page                                                                                                                                ii

Declaration                                                                                                                              iii

Certification                                                                                                                            iv

Dedication                                                                                                                              v

Acknowledgments                                                                                                                  vi

Table of Contents                                                                                                                   vii

List of Tables                                                                                                                          xi

List of Plates                                                                                                                           xii

Abstract                                                                                                                                  xiii


1.1      Background of study                                                                                                   1

1.2      Statement of Problems                                                                                                 2

1.3      Justification                                                                                                                  3

1.4      Objective of the study                                                                                                             3


2.1       General overview of spices                                                                                         4

2.2       Values of spices                                                                                                          5

2.2.1    Health benefits of Spices                                                                                            9  Cardiovascular disease                                                                                               9  Cancer                                                                                                                        12  Mental health and cognition                                                                                      14 Type 2 diabetes mellitus                                                                                             16 Osteoarthritis and inflammatory response                                                                  18

2.2.2    Dietary implications of spices                                                                                     19 Food variety and diversity                                                                                          19 Herbs and spices Increase bioavailability of other nutrients                                       20

2.2.3    Antioxidant properties of traditional spices                                                               21

2.3       Cocoplum (Chysobalanus icaco)                                                                                22

2.3.1    Description of cocoplum seed (Chysobalanus icaco)                                                 23

2.3.2    Scientific classification                                                                                               25

2.4      African Black Pepper (Piper guineense)                                                                      26

2.4.1     Description of African black pepper (Piper guineense)                                             26

2.4.2    Uses of African black pepper (Piper guineense}                                                        26

2.4.3    Nutritional composition of African black pepper (Piper guineense)                          28

2.4.4    Antioxidating & preservative properties of African black pepper (Piper guineense)29

2.4.5    Phytochemistry of African black pepper (Piper guineense)                                       30

2.4.6    Scientific classification of African black pepper (Piper guineense)                           31

2.5       Aidan Fruits (Tetrapleura tetraptera)                                                                         31

2.5.1    Description of Aidan fruit (Tetrapleura tetraptera)                                                   32

2.5.2    Uses of Aidan fruit (Teirapleura tetraptera)                                                              33

2.5.2    Nutritional composition of Aidan fruit (Tetrapleura tetraptera)                                34

2.5.3    Scientific classification of Aidan fruit (Tetrapleura tetraptera)                                 35

2.6       Bioactive Ingredients                                                                                                  35

2.6.1    Flavonoids                                                                                                                  36 Benefits and uses of flavonoids                                                                                 37

2.7       Carotenoids (Pro-Vitamin A)                                                                                     38

2.8.      Tannin                                                                                                                         40

2.8.1    Benefits and Uses of tannin                                                                                       41 CHAPTER 3: MATERIALS AND METHODS

3.1       Materials/Sources of Materials                                                                        45

3.2       Sample Preparation                                                                                                     46

3.2.1    Sample Formulation                                                                                                    46

3.3       Proximate Determination                                                                                            48

3.3.1     Determination of protein content                                                                              48

3.3.2    Fat content determination                                                                                           49

3.3.3    Total Ash determination                                                                                             49

3.3.4    Crude Fiber determination                                                                                          50

3.3.5    Moisture Content determination                                                                                 50

3.3.6    Carbohydrate determination                                                                                                   51

3.4       Vitamin and Mineral Content Determination                                                             52

3.4.1    Determination of Pro - vitamin A (Carotene)                                                             52

3.4.2    Determination of vitamin C                                                                                        53

3.4.3    Determination of Thiamine (Vitamin Bi)                                                                    53

3.4.4    Determination of Riboflavin (Vitamin B2)                                                                 54

3.5       Determination Mineral Content                                                                                  54

3.5.1    Determination of phosphorus content                                                                        54

3.5.2    Determination of calcium and magnesium                                                                 55

3.5.3    Determination of Sodium, Zinc, Iron, Manganese and Potassium Contents            56

3.6       Determination of Bioactive Ingredients (Phytochemicals)                                         57                               

3 .6. 1  Determination of tannin contents                                                                               57

3.6.2    Determination of flavonoid contents                                                                          58       

3.6.3    Determination of alkaloid contents                                                                            58                                                                   

3.6.4    Determination of trypsin inhibitor contents                                                                59                                                  

3.6.5    Determination of Saponin contents                                                                            61

3.6.6    Determination of steroid contents                                                                              61

3.6.7    Determination of phenol contents                                                                              62

3.6.8    Determination of terpenoid contents                                                                          63

3.7       Determination of Antioxidant Contents                                                                     63

3.7.1    DPPH Radical Scavenging Activity                                                                           63

3.7.2    Ferric Reducing Antioxidant Power Assay (FRAP)                                                  64

3.8       Statistical Analysis                                                                                                      64


4.1       Proximate Composition of the Spice Samples                                                            65

4.1.1    Moisture composition of the spice samples                                                                65

4.1.2    Protein composition of the spice samples                                                                   67

4.1.3    Crude fibre composition of the spice samples                                                            68

4.1.4    Fat composition of the spice samples                                                                         69

4.1.5    Ash composition of the spice samples                                                                        70

4.1.6    Carbohydrate content of the spice                                                                              71

4.2       Mineral Composition of the Spice Samples                                                                72

4.2.1    Phosphorus composition of the spice                                                                          74

4.2.2    Calcium composition of the spice samples                                                                 75

4.2.3    Magnesium composition of the spice                                                                          75

4.2.4    Zinc content of spices                                                                                                 76

4.2.5    Iron content of spices                                                                                                 77

4.2.6    Potassium content of spices                                                                                        78

4.2.7    Manganese content of spices                                                                                      79

4.3 Vitamin content of spices, Cocoplum, African Black Pepper and Aidan Fruit blends    80

4.3.1    Pro - vitamin A content of spices                                                                               80

4.3.2    Thiamin content of spices                                                                                           83

4.3.3    Riboflavin content of the spices                                                                                 83

4.3.4    Ascorbic acid content of the spices                                                                            84

4.4       Bioactive Compounds of Spices; Cocoplum, African Black Pepper and Aidan

            Fruit Blend                                                                                                                  85

4.4.1    Tannin content of the spices                                                                                       86

4.4.2    Flavonoid content of the spices                                                                                  88

4.4.3    Alkaloid content of the spices                                                                                    88

4.4.4    Phenol content of the spices                                                                                                   89

4.4.5    Saponin content of the spices                                                                                     90

4.4.6    Steroid content of the spices                                                                                      91

4.4.7    Terpenoid content of the spices                                                                                  92

4.4.8    Trypsin inhibitor content of the spices                                                                        93

4.5       Antioxidant Properties of Spices; Cocoplum, African Pepper, and Aidan

            Fruit Blends                                                                                                                94

4.5.1     FRAP radical scavenging ability of the spices                                                          94

4.5.2    DPPH radical scavenging ability of the spices                                                           97 CHAPTER 5: CONCLUSION AND RECOMMENDATIONS

5.1       Conclusion                                                                                                                  173

5.2       Recommendation                                                                                                        174

References                                                                                                                  175

Appendices                                                                                                                 200 






2.1  Binomial nomenclature of Cocoplum (Chysobalanus icaco)                                           25


2.2  Binomial nomenclature of African Black Pepper (Piper guineense)                               31


2.3  Binomial nomenclature of Aidan fruit (Tetrapleura tetraptera)                          35


3.1: Mixture formulations of spice blends from Mixture Design                                           47


4.1: The comparative nutritional composition of spices; Cocoplum

        (Chysobalanus icaco), African black pepper (Piper guineense)

        and Aidan fruit (Tetrapleura tetraptera) blends                                                             66                                                                                       


4.2: Mineral composition of spices; Cocoplum (Chysobalanus icaco),

       African black pepper (Piper guineense) and Aidan fruit

        (Tetrapleura tetraptera) blends                                                                                       74


4.3: Vitamin composition of spices; Cocoplum (Chysobalanus icaco),

        African black pepper (Piper guineense) and Aidan fruit

        (Tetrapleura tetraptera) blends                                                                           82


4.4: Bioactive compounds of spices; Cocoplum (Chysobalanus icaco),

        African black pepper (Piper guineense) and Aidan fruit

        (Tetrapleura tetraptera) blends                                                                                       87


4.5: Antioxidant properties of spices; Cocoplum (Chysobalanus icaco),

        African pepper (Piper guineense) and Aidan fruit (Tetrapleura tetraptera)

        Blends African black                                                                                                      96












1: Cocoplum (Chysobalanus icaco)                                                                            45


2: African black pepper (Piper guineense)                                                                  45


3: Aidan fruit (Tetrapleura tetrapterd)                                                                       45


4: Blend of cocoplum, African black pepper and Aidan fruits spices                        46











Bioactive ingredients/compounds are types of chemicals found in small amount in plants such as fruits, vegetables, nuts, seeds, oils and whole grains that provide health benefits beyond the basic nutritional value (Gokmen, 2016). These compounds have actions in the body that may present therapeutic potential with influence on energy intake, reducing pro-inflamatory conditions, oxidative stress and metabolic disorders (Siriwardhana et al., 2013).They otherwise promote good health. There are several bioactive ingredients in both plants and animals,however, studies indicate that consuming foods that are rich in bioactive ingredients at a higher rate have positive effect on humans and could prevent risk of numerous diseases such as cancer,heart disease,stroke,diabetes,cataracts and age related functional problems (Hassimotto et al, 2009).Some of these bioactive ingredients include lycopene, tannins, flavonoids, carotenoids, phytosterols, steroids, phenols and polyphenols, anthocyanins, alkaloids, terpenes etc.These compounds (bioactive ingredients) according to (Carbonell-Capella et al; 2014) are capable of modulating metabolic processes and show positive properties such as antioxidant effect, inhibition of receptor activities, inhibition of enzymes and induction of gene expression. These ingredients could as well be found in most of our indigenous spices, if not all.

Spices as the name implies, according to FDA, (2015) are aromatic vegetable substances, in whole, broken or ground form whose significant function in food is seasoning rather than nutrition. Spices are mainly used to improve the sensory characteristics of foods and beverages (Gunasekar et al., 2012).However, most of the common spices are usually extracted from plants grown in warm or tropical climates while herbal plants can be grown in nearly all conditions and geographical areas around the world,spices have more intense taste as used in small amount (Ranton,2004).They are known to have beneficial effects for humans including digestive stimulant, anti-infamatory, antimicrobial, antioxidant and anticarcinogenic activities mostly attributed to the polyphenols and other constituents which contribute to the plant's biological activity resulting in benefits for human health and this health benefits are mediated by their ability to prevent free radical formation, remove radicals, repair oxidative damage, eliminate damaged molecules by their antioxidant ability (Gordon,1996).The spice antioxidant activity is related to variety of chemical compounds. The most effective antioxidants act by interrupting the free chain reaction. These compounds donate H+ radicals to the free radicals formed during oxidation becoming radicals themselves that are stabilized by the resonance delocalization of the electron within the aromatic ring(Nawar,1996).Many of the phenolics also lack positions suitable for molecular oxygen attack or quench free radical oxygen (Brewer,2011).


There is little or no knowledge in the comparartive assessment of the nutrient composition and bioactive ingredient levels of herbs and spices as they are only seen from functional food perspective and usage in diet. The use of these indigenous and non-indigenous spices are      normally inherited from tradition which vary from community to community in Africa as such, it is paramount to establish the nutrient, bioactive ingredient compositions of these spices as this will help to make bettwe use of them in promoting health of consumers. Nutrient composition, bioactive activity and antioxidant activities of Cocoplum, African black pepper and Aidan fruit blends are paramount for their effective utilization.                                 


Considering the problems posed on human health, studies on nutrient composition, bioactive and antioxidant properties of these commonly used  Cocoplum (Chysobalanus icaco), African black pepper (Piper guineese) and Aidan fruit(Tetrapleura tetraptera) blends will bring out their real value in food and even medicinal application. This study is not only expected to provide comparative assessment to consumers of the spices on bioactive and antioxidant activities but also create general awareness of their nutritional potentials.  This would form a basis for improved utilization of these spices as food ingredients. The information will also help food professionals to develop of novel products. The information will also create awareness and encourage the use of these spices at local and industrial levels.


The main objective of this work is to comparatively assess, the bioactive ingredients of three Nigerian spices; Cocoplum (Chysobalanus icaco), African black pepper (Piper guineese) and Aidan fruit (Tetrapleura tetraptera) blends.

 The specific objectives are to:

                    i.            Formulation of the blends

           ii.            Determine the proximate composition of Cocoplum (Chysobalanus icaco), African black pepper (Piper guineese) and Aidan fruit (Tetrapleura tetraptera) blends.

                iii.            Determine the mineral and vitamin contents of Cocoplum (Chysobalanus icaco), African black pepper(Piper guineese) and Aidan fruit (Tetrapleura tetraptera) blends.

                iv.            Determine the bioactive ingredients of Cocoplum (Chysobalanus icaco),African black pepper(Piper guineese) and Aidan fruit(Tetrapleura tetraptera) blends.

                  v.            Assess the antioxidant activities of the spice blends.


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