PHYTOCHEMICAL AND ANTIMICROBIAL STUDIES OF THE METHANOL EXTRACT OF THE ROOT OF NAPOLEONAEA HEUDELOTII (A.JUSS)

PHYTOCHEMICAL AND ANTIMICROBIAL STUDIES OF THE METHANOL EXTRACT OF THE ROOT OF NAPOLEONAEA HEUDELOTII (A.JUSS)

Abstract

The extracts of the foundation a part of Napoleonaea heudelotii have been subjected to phytochemical and anti-microbial studies. Extraction become performed through non-stop Soxhlet extraction the usage of methanol. The phytochemical screening of the crude methanol extract, chloroform and ethyl acetate fractions found out the presence of carbohydrate, cardiac glycosides, saponins, steroids, triterpenes, flavonoids and tannins. The end result of the antimicrobial screening of the crude methanol extract, ethyl acetate and chloroform fractions confirmed interest in opposition to Staphylococcus aureus, Streptococcus pyogenes, Bacillus subtilis, Escherichia coli, Salmonella typhi, Pseudomonas aeruginosa, Proteus vulgaris, and Candida albicans. However, the chloroform fraction become the maximum energetic fraction in opposition to the check microorganisms. The area of inhibition of the methanol extract ranged among sixteen mm and 21 mm, the chloroform fraction ranged among 17 mm and 25 mm at the same time as the ethyl acetate fraction ranged among 15 mm and 21 mm. The MIC outcomes of methanol extract, ranged among 12.five mg/ml and 1.562 mg/ml, chloroform fraction ranged among 12.five mg/ml and 1.562 mg/ml, at the same time as ethyl acetate ranged among 6.25 mg/ml and 1.625 mg/ml. The MBC of methanol extract and chloroform fraction ranged among 12.five mg/ml and 1.562 mg/ml, at the same time as that of ethyl acetate fraction ranged among 6.2 mg/ml and 1.562 mg/ml. The chloroform fraction being the maximum energetic fraction become subjected to giant chromatographic purification; white crystalline strong labelled NHPE have been remoted. The systems of the remoted compounds have been decided to be a combination α-amyrin and β-amyrin the usage of 1D and 2D NMR.

TABLE OF CONTENTS

Title Page
Abstract
Table of Contents
List of Abbreviations

CHAPTER ONE
1.zero INTRODUCTION
1.1 Secondary Metabolites
1.1.1 Alkaloids
1.1.2 Flavonoids
1.1.three Terpenes
1.1.four Steroids
1.1.five Saponins
1.1.6 Tannins
1.1.7 Glycosides
1.three Justification
1.four Aim and Objectives

CHAPTER TWO
2.zero LITERATURE REVIEW
2.1 Botanical Description of Napoleonaea Species
2.1.1 Taxonomy of Napoleonaea Species
2.2 Origin and geographical distribution
2.three Medicinal makes use of
2.four Phytochemical Constituents of Napoleonaea species
2.five Pharmacological Activities of Napoleonaea species
2.6 Scientific Classification of Napoleonaea heudelotti

CHAPTER THREE
three.zero MATERIALS AND METHODS
three.1 Materials
three.1.1 Solvents
three.1.2 Apparatus
three.1.three Reagents for phytochemical screening
three.1.four Test microorganisms used
three.1.five Materials used for chromatographic strategies
three.2 Collection of the Plant Material
three.three Extraction of the Plant Material
three.four Phytochemical Screening
three.four.1 Test for glycosides
three.four.2 Test for cardiac glycoside
three.four.three Test for tannins
three.four.four Test for saponins
three.four.five Test for flavonoids
three.four.6 Test for carbohydrates
three.four.7 Test for blended decreased sugar
three.four.eight Test for steroids/terpenoids
three.four.nine Test for alkaloids
three.five Antimicrobial Screening
three.6 Purification of chloroform fraction
three.6.1 Thin layer chromatography
three.6.2 Preparation of preparative TLC
three.6.three Isolation of the natural compound
three.6.four Melting factor willpower
three.6.five Spectra Analysis

CHAPTER FOUR
four.zero RESULTS
four.1 Percentage ( %) Recovery on Extraction
four.2 Phytochemical Screening of extract and fractions
four.three Antimicrobial Susceptibility check
four.four Zone of Inhibition (mm)
four.five Minimum inhibitory concentration (MIC) of extract and fractions
four.6 Minimum bactericidal concentration (MBC) of extract and fractions
four.7 Preparative skinny layer chromatography
four.eight Chemical check of the remoted compound
four.nine Spectroscopic evaluation of pattern NHPE

CHAPTER FIVE
5.0 DISCUSSION

CHAPTER SIX
6.0 SUMMARY, CONCLUSION, AND RECOMMENDATION
6.1 Summary
6.2 Conclusion
6.3 Recommendation
REFERENCES

List of Abbreviations

PPM Part per million

NMR Nuclear magnetic resonance

TLC Thin layer chromatography

DEPT Distortionless Enhancement by Polarization Transfer

MIC Minimum Inhibitory Concentration

MBC Minimum Bactericidal Concentration

1H NMR Proton Nuclear Magnetic Resonance

13C NMR Carbon Nuclear Magnetic Resonance

NHPE Napoleonaea heudelotii plant extract

CHAPTER ONE

1.0 INTRODUCTION

Medicinal plants have been identified and used throughout human history. Plants have the ability to synthesize a wide variety of chemical compounds that are used to perform important biological functions, and to defend against attack from predators such as insects, fungi and herbivorous mammals (Babalola, 2009). Chemical compounds in plant mediate their effects on the human body through processes identical to those already well understood for the chemical compounds in conventional drugs; thus herbal medicines do not differ greatly from conventional drugs in terms of how they work. This enables herbal medicines to be as effective as conventional medicines, but also gives them the same potential to cause harmful side effects. Ethnobotany (the study of traditional human uses of plants) is recognized as an effective way to discover future medicines. In 2001, researchers identified 122 compounds used in modern medicine which were derived from ethnomedical plant sources (Babalola, 2009). Many of the pharmaceuticals currently available to physicians have a long history of use, as herbal remedies, including aspirin, digitalis, quinine, and opium. Treatment of diseases is almost universal among non-industrialized societies, and is often more affordable than purchasing expensive modern pharmaceuticals (Beltrame et al., 2002). The World Health Organization (WHO) estimates that 80 percent of the population of some Asian and African countries presently use herbal medicine for some aspect of primary health care (Beltrame et al., 2002). Studies in the United States and Europe have shown that the use of herbal medicine is less common in clinical settings, but has become increasingly more in recent years as scientific evidence about it effectiveness has become more widely available. The annual global export value of pharmaceutical plants in 2011 accounted for over US$ 2.2 billion. Plants have continued to be major source of medicine either in the form of traditional medicine preparations or as pure active principles (Hill, 2011). This has made it important to identify plants with useful therapeutic actions for possible isolation and characterization of their active constituents. About 80 % of the world population relies on the use of traditional medicine which is predominantly based on plant materials (Brunton et al., 2006). Plant have been part of our lives since beginning of time, we get numerous products from plants, most of them, not only good and beneficial but also crucial to our existence. The use of plant to heal or combats illness is probably as old as human kind. Out of these simple beginning came the pharmaceutical industry. Yet the current view of plant is very different from how it all started. The acceptance of traditional medicine as an alternate form of health care and the development of microbial resistance to the available antibiotics has led researchers to investigate the antimicrobial herbal extract (WHO, 1993). In Africa, particularly Nigeria is rich in plants which are used in herbal medicine to cure diseases and to heal injuries. Some of these plants exhibit a wide range of biological and pharmacological activities such as antihelmenthics, oxytoxic laxative (Hostettmann et al., 2012). The secondary metabolites of plant provide human with numerous biological active components which have been used extensively as drugs, foods, additives, flavours, insecticides and chemicals. They exhibited remarkable biological activities, which include inhibitory effects on enzymes, modulatory effects on some cell types, protect against allergies antioxidants (Dongmo et al., 2001).

1.2 Secondary metabolism

1.2.1 Alkaloids

Alkaloids are a group of naturally occurring chemical compounds containing mostly basic nitrogen atoms. B. cornine (1) and quinine (2). This group also includes some related compounds with neutral and weakly acidic properties. Some synthetic compounds of similar structure are also attributed to alkaloids. In addition to carbon, hydrogen and nitrogen, alkaloids may contain oxygen, sulfur and, rarely, other elements such as chlorine, bromine and phosphorus. Alkaloids are produced by a wide variety of organisms, including bacteria, fungi, plants, and animals, and belong to a group of natural products (also called secondary metabolites). Many alkaloids can be purified from crude extracts by acid-base extraction. Many alkaloids are toxic to other organisms (Kumar et al., 2010). They often have pharmacological effects and are used in medicine, recreational medicine, or theological rituals (Kumar et al., 2010).

1.2.2 Flavonoids

Flavonoids are a class of plant secondary metabolites. Flavonoids have also been described as non-ketone polyhydroxypolyphenolic compounds, more specifically called flavonoids. B. Isoflavans (3) and neoflavonoids (4). The three rings or heterocycles of the flavonoid skeleton are commonly referred to as the A, B, and C rings. Ring A typically shows a phloroglucine substitution pattern. Flavonoids are ubiquitous in plants and serve many functions (Dongmo et al., 2001). Flavonoids are the main plant pigments for flower coloring, producing yellow or red/blue pigmentation in petals to attract pollinators.In higher plants, flavonoids are UV-filtering, symbiotic nitrogen-fixing , and is involved in flower pigmentation. They also function as chemical messengers, physiological regulators, and cell cycle inhibitors (Kumar et al., 2010).

1.2.3 Terpenes

Compounds classified as terpenes constitute arguably the largest and most diverse class of natural products. Most of these compounds are found only in plants, but some of the larger, more complex terpenes are found in animals. B. Isopentenyl pyrophosphate (5) is the basic unit by which terpenes are found in natural organisms. Instead, carbon number and structural organization are the key features. Terpenes can be assumed to be composed of isoprene (specifically isopentane) units, an empirical property known as the isoprene rule (Dongmo et al., 2001). 1.2.4 Steroids

An important class of lipids called steroids, which are actually metabolic derivatives of terpenes, are usually treated as a separate group. recognizable by structure. These rings are synthesized by biochemical processes from the cyclization of a 30-carbon chain. Animals, fungi and plants contain hundreds of steroids. B. Cholesterol (6), sex hormones, estradiol, and testosterone (Kaisar et al., 2011).

1.2.5 Saponins

Saponins are a class of compounds found in many plant species. More specifically, they are amphiphilic glycosides, grouped by soap-like foams produced when shaken in aqueous solutions, and structurally combined with lipophilic triterpene derivatives to form one or more are classified by having a hydrophilic glycoside moiety of (Kaisar et al., 2011).

Although saponins were historically thought to be of plant origin, they have also been isolated from marine organisms. Saponins are actually found in many plants, and their name comes from the soapwort plant (Saponaria genus, Caryophyllaceae), whose roots were historically used as soaps. Saponins are also found in the plant family

Sapindaceae with its distinct genus Sapindaceae (soapberries or soapnuts) and closely related families Saccharaceae (maple) and Birchaceae. An example of a saponin is solanine (7). Gynostemma pentaphyllum (Gynostemma, Cucurbitaceae) contains a lot of gypenosides, and Korean ginseng or red ginseng (Panax, Araliaceae) contains a lot of ginsenosides. Within these families are found chemical compounds of this class.

1.2.6 Tannins

Tannins are astringent, bitter plant polyphenolic compounds that bind and precipitate proteins and a variety of other organic compounds, including amino acids and alkaloids.

The term tannin (from the Old High German word tanna, which means oak or fir, like tannabaum), refers to the use of oak wood tanning agents in tanning animal hides into leather. That is why the terms “tanning” and “tanning” are used to describe the treatment of leather. However, the term ‘tannin’ is broadly applied to large polyphenolic compounds containing sufficient hydroxyl groups and other suitable groups (such as carboxyl groups) to form strong complexes with various macromolecules (Kaisar et al., 2011).

Tannin compounds (such as gallic acid (8)) are widely distributed in many plant species and play a role in protecting against predators and possibly as insecticides and in regulating plant growth. Tannin astringency causes a dry, shriveled mouthfeel after drinking unripe fruit or red wine. Similarly, the destruction or alteration of tannins over time plays an important role in fruit ripening and wine aging (Kaisar et al., 2011). .

1.2.7 Glycosides

Glycosides are sugar compounds attached to non-carbohydrate groups. Used to treat congestive heart failure and arrhythmia. An example of a glycoside is salicin (9). These glycosides are found as secondary metabolites in some plants and some insects (Kaisal et al., 2011).

1.3 Presentation of research topic

There are many different types of drugs used to treat different types of infections, but few are known to researchers. Therefore, we need to know more about these compounds that are effective against these diseases.

1.4 Legitimacy of research

Based on the ethnomedical claims of Napoleonaea heudelotti, scientific research is needed to identify the active ingredient responsible for these effects.

1.5 Purpose

Isolation and characterization of bioactive compounds that may be present in the root part of Napoleonaea heudelotii and determination of their activity against common microorganisms

1.6 Purpose

The objectives of this research work are achieved through the following objectives:

of. Identify the phytochemical constituents present in the root part of plants.

b. isolate some of the compounds present in plant roots

c. Assess the antibacterial activity of plant root parts and isolated compounds

spectroscopic techniques are used to elucidate the structures of isolated compounds.