PHYTOCHEMICAL ANALYSIS OF THIN LAYER CHROMTOGRAPHIC STUDY AND U.V ANALYSIS OF Ocimum gratissimum

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Product Code: 00005995

No of Pages: 35

No of Chapters: 1 - 5

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Table of Content


CHAPTER ONE     

1.0 INTRODUCTION         

1.1 Background of the study           

1.2 STATEMENT OF PROBLEM

1.3 JUSTIFICATION OF THE STUDY  

1.4 AIM         

1.5 OBJECTIVES OF THE STUDY     

   

CHAPTER TWO    

2.0 LITERATURE REVIEW        

2.1. TRADITIONAL USES O. gratissimum         

2.2. ALTERNATIVE AND COMPLEMENTARY MEDICINAL       

2.3 OCIMUM SPECIES    

 

CHAPTER THREE

3.0 Materials and Methods  

3.1 MATERIALS    

3.2 METHODOLOGY        

3.2.1 Preparation of the Extract

3.2.2 Preliminary Phytochemical Screening          

3.2.3 Test for Alkaloids        

3.2.4 Test for Flavonoids     

3.2.5 Test for Anthraquinones        

3.2.6 Test for Saponins        

3.2.7 Test for Steroids and Triterpenes      

3.2.8 Salkowski test: 

3.2.9 Test for Tannins          

3.2.10 Test for Carbohydrates        

3.2 Test for Phenols 

3.2.1 Test for Glycosides      

3.2.2 Thin-layer Chromatographic Studies (TLC)

3.2.3 UV spectroscopic analysis     

 

CHAPTER FOUR   

4.0 RESULTS AND DISCUSSION          

 

CHAPTER FIVE    

5.0 SUMMARY, RECOMMENDATION AND CONCLUSION          

5.1 SUMMARY

5.2. RECOMMENDATION           

5.3 CONCLUSION  

References     

ABSTRACTS

 

The Phytochemical analysis, thin-layer chromatographic study and U.V analysis of Ocimum gratissimum was carried out. Ocimum gratissimum is a variable, polymorphic species with many forms. It has at times been treated as a number of distinct species, in particular Ocimum suave Willd., and Ocimum viride Willd., have been recognized as distinct in many treatments. However, it is difficult to define clear differences between these three species because variations between them form a continuum merging from one to another. All three are treated here as part of the one complex species Ocimum gratissimum. The plant was collected from Jigawa state polytechnic, Dutse, the sample was extracted from methanol, hexane and chloroform for both qualitative and quantitative phytochemical analysis. The analysis shows the presence of all the phytochemicals in all the three solvents Alkaloid, Saponin, carbohydrate, tannin, glycoside, flavonoid, anthraquinones, phenols steroids and triterpenes were found to be present in all the three solvents.  however, anthraquinones and steroids were absent in methanol extract while carbohydrates and steroids were absent in hexane extract. carbohydrates, steroids and    terpenes were also found to be absent in the chloroform extract excepts steroids and carbohydrate which are absent in all the solvent and anthraquinones which is found to be absent in methanol extract.  The thin layer chromatographic analysis reveals the presence of only one pale green spot in hexane, two green and yellow spots in Hexane and two green and pink spots in chloroform extracts. The result of the U.V spectroscopy of Occinum gratissinum at various wavelength was carried out, 400nm was studied to be the highest peak with higher activity.






                                                   CHAPTER ONE


1.0 INTRODUCTION

1.1 Background of the study

Organic chemistry is an art executed science. The art of synthesis and isolation of natural products and bioactive compounds makes it one of the most interesting and finest areas of modern chemistry. The key role played by plant-based systems in the healthcare of different cultures has been extensively documented, and the World Health Organization (WHO) has estimated that approximately 65-80% of the world’s population relies mainly on plant-derived traditional medicines for their primary health care. Most of these natural products are secondary plant metabolites. Keeping this in view, we felt an urge to synthesize some natural products and test their bioactivity and also planned for the isolation of some bioactive natural compounds. Natural products have played a key role in health care and prevention of diseases for the past thousands of years. Several ancient civilizations, such as Indians, Chinese, and North Africans have provided written evidence for the use of natural sources for curing various ailments. The importance of natural products as anticancer agents can be seen between the years 1981- 2006, where about a hundred anticancer agents have been developed, of which, nine were pure natural products, eleven were derived from a natural product pharmacophore, eighteen were natural product mimics, and twenty-five were natural product derivatives. Thus, making the natural sources as significant contributors to the health care system. Ocimum gratissimum locally called as Nimma tulasi is widespread in India and South Africa and also grows across tropical regions of the globe. It is a traditional herb with a wide array of phytochemical constituents and has been reported for its diverse physiological properties. Earlier reports showed that O. gratissimum possess a wide range of phytochemicals such as flavonoids, polyphenols and volatile compounds like eugenol, thymol, and geraniol Its medicinal uses include vasorelaxation, anti-inflammatory, anti myco toxigenic and antioxidant activities. Plants show enormous versatility in synthesizing complex materials which have no immediate obvious growth or metabolic functions. These complex materials are referred to as secondary metabolites. Plants secondary metabolites have recently been referred to as phytochemicals (Farath.,2014). Phytochemicals are naturally occurring and biologically active plant compounds that have potential disease inhibiting capabilities. It is believed that phytochemicals may be effective in combating or preventing disease due to their antioxidant effect. Antioxidants protect other molecules (in vivo) from oxidation when they are exposed to free radicals and reactive oxygen species which have been implicated in the aetiology of many diseases and in food deterioration and spoilage. Medicinal plants have been used for centuries before the advent of orthodox medicine. Leaves, flowers, stems, roots, seeds, fruit, and bark can all be constituents of herbal medicines. The medicinal values of these plants lie in their component phytochemicals, which produce definite physiological actions on the human body. The most important of these phytochemicals are alkaloids, tannins, flavonoids and phenolic compounds Ocimum gratissimum Linn (Labiatae) is grown for the essential oils in its leaves and stems. Eugenol, thymol, citral, geraniol and linalool have been extracted from the oil. Essential oils from the plant have been reported to possess an interesting spectrum of antifungal properties. The antinociceptive property of the essential oil of the plant has been reported. The whole plant and the essential oil are used in traditional medicine especially in Africa and India. The essential oil is also an important insect repellent (Afolabi ., et al. 2010.) O. gratissimum is germicidal and has found wide use in toothpastes and mouth washes as well as some topical ointments. It is used as an excellent gargle for sore throats and tonsillitis. It is also used as an expectorant and a cough suppressant. The plant extract is used against gastrointestinal helminths of animals and man. In addition, O. gratissimum carminative properties make it a good choice for upset stomach. It is used as an emetic and for hemorrhoids. The plant is also used for the treatment of rheumatism, paralysis, epilepsy, high fever, diarrhoea, sunstroke, influenza, gonorrhoea and mental illness. In addition, the plant is used as a spice and condiment in the southern part of Nigeria. The plant is found throughout the tropics and subtropics and its greatest variability occurs in tropical Africa and India. In South East Asia, it is cultivated as a home garden crop but it is grown on a commercial scale in Vietnam. It is used for a variety of reasons. In culinary, it is used in salads, soups, pastas, vinegars and jellies in many parts of the world. The Thai people are popularly known to use it in food flavouring. In traditional medicine, the leaves have been used as a general tonic and anti-diarrhea agent and for the treatment of conjunctivitis by instilling directly into the eyes; the leaf oil when mixed with alcohol is applied as a lotion for skin infections and taken internally for bronchitis. The dried leaves are snuffed to alleviate headaches and fever among other uses. Although, conventional antibiotics have been very useful in orthodox medicine, it has been argued by many that its concomitant use with herbal extracts is not desirable as one normally antagonizes the activity of the other. Considering the fact that Ocimum gratissimum is used in most local dishes/foods to achieve a variety of purposes, there is need to ascertain if its extract antagonizes or acts as a synergy when used together with conventional antibiotics. In addition, despite the fact that the various extracts of O. gratissum have been tested in vitro and shown to be active against some bacterial and fungal isolates, specific strain differences supposes that a lot more strains of bacteria and fungi across other regions be tested to ascertain their in vitro activity against this spice and this was one of the motivations for our study. This study thus investigated the antibacterial and antifungal properties of O. gratissimum ethanolic leaf extract and its interaction with some disc antibiotics.

The ethanolic extract of the leaves of Ocimium gratisimum L. (Lamiaceae), used in traditional medicine for the treatment of several ailments such as urinary tract, wound, skin and gastrointestinal infections, was evaluated for its antibacterial properties against four clinical bacteria isolates namely: Escherichia coli, Proteus mirabilis, Staphylococcus aureus and Pseudomonas aeruginosa and the antifungal properties using a clinical isolate of Candida albicans. A typed bacterium of Escherichia coli ATCC 11775 and another typed fungal strain of Candida albicans (ATCC 90028) were also included. The study also intended to verify if the concomitant administration of conventional antibiotics with Ocimium gratisimum which is normally taken as food (spice) will negatively affect its activity.

 

 Organic chemistry is an art executed science. The art of synthesis and isolation of natural products and bioactive compounds makes it one of the most interesting and finest areas of modern chemistry.

The key role played by plant-based systems in the healthcare of different cultures has been extensively documented, and the World Health Organization (WHO) has estimated that approximately 65-80% of the world’s population relies mainly on plant-derived traditional medicines for their primary health care.

Most of these natural products are secondary plant metabolites. Keeping this in view, we felt an urge to synthesize some natural products and test their bioactivity and also planned for the isolation of some bioactive natural compounds.

Natural products have played a key role in health care and prevention of diseases for the past thousands of years. Several ancient civilizations, such as Indians, Chinese, and North Africans have provided written evidence for the use of natural sources for curing various ailments.

The importance of natural products as anticancer agents can be seen between the years 1981-2006, where about a hundred anticancer agents have been developed, of which, nine were pure natural products, eleven were derived from a natural product pharmacophore, eighteen were natural product mimics, and twenty five were natural product derivatives.

Thus making the natural sources as significant contributors to the health care system.

Ocimum gratissimum locally called as Nimma tulasi is widespread in India and South Africa and also grows across tropical regions of the globe. It is a traditional herb with a wide array of phytochemical constituents and has been reported for its diverse physiological properties. Earlier reports showed that O. gratissimum possess a wide range of phytochemicals such as flavonoids, polyphenols and volatile compounds like eugenol, thymol, and geraniol Its medicinal uses include vasorelaxation, anti-inflammatory, anti mycotoxigenic and antioxidant activities.

Plants show enormous versatility in synthesizing complex materials which have no immediate obvious growth or metabolic functions. These complex materials are referred to as secondary metabolites. Plants secondary metabolites have recently been referred to as phytochemicals [1].

Phytochemicals are naturally occurring and biologically active plant compounds that have potential disease inhibiting capabilities. It is believed that phytochemicals may be effective in combating or preventing disease due to their antioxidant effect. Antioxidants protect other molecules (in vivo) from oxidation when they are exposed to free radicals and reactive oxygen species which have been implicated in the aetiology of many diseases and in food deterioration and spoilage. Medicinal plants have been used for centuries before the advent of orthodox medicine he genus Ocimum is a member of the Lamiaceae family, comprising more than 150 species. It grows widely and is distributed throughout temperate regions of the world with the greatest number of species in Africa. The best known species are the strongly aromatic herb Ocimum basilicum (Thai basil) and Ocimum gratissimum (African basil) as well as the medicinal herb Ocimum tenuiflorum, also known as Ocimum sanctum (holy basil or tulsi in Hindi). Basil is the main ingredient of pesto sauce but is also used to flavor other sauces and soups. Different parts including the leaves, stems, flowers, roots, seeds, and even the whole plant are useful. The seeds are edible, and when soaked in water become mucilaginous. The leaves can be eaten as a salad. Basil is widely used in traditional medicine. It is used in Ayurveda and in traditional Chinese medicine for treating digestive system disorders, such as stomach ache and diarrhoea, kidney complaints, and infections.  Ocimum gratissimum is herbaceous plant belonging to Lamiaceae family. The plant is indigenous to tropical areas especially India and West Africa. It is known by different names in various parts of the world. In India, the most commonly used ones being Vriddhu tulasi (Sanskrit), Ram tulasi (Hindi), Nimma tulasi (Kannada). Ocimum gratissimum has been used extensively in the traditional system of medicine in several countries. It is used for medicinal, condiment and culinary purpose. The flowers and the leaves of this plant are rich in essential oils so it is used in preparation of tea and infusion. The plant is commonly used in folk medicine to treat different diseases such as upper respiratory tract infections, diarrhoea, headache, eye and skin diseases, pneumonia, cough, fever and conjunctivitis. In India, the whole plant has been used for the treatment of sunstroke, headache and influenza as a diaphoretic, antipyretic and for its anti-inflammatory activity. The infusion of Ocimum gratissimum leaves is used as pulmonary antisepticum, antitussivum and antispasmodicum. Since no work has so far been cited on Ocimum gratissimum root, the present study deals with the investigation of medicinally useful bioactive phytoconstituent in root extracts and their antimicrobial activity along with total phenolic content, (Alfred .,2017).

1.2 STATEMENT OF PROBLEM

The ethanolic extract of the leaves of Ocimium gratisimum, used in traditional medicine for the treatment of several ailments such as urinary tract, wound, skin and gastrointestinal infections, was evaluated for its antibacterial properties against four clinical bacteria isolates namely: Escherichia coli, Proteus mirabilis, Staphylococcus aureus and Pseudomonas aeruginosa and the antifungal properties using a clinical isolate of Candida albicans. A typed bacterium of Escherichia coli and another typed fungal strain of Candida albicans were also included. The study also intended to verify if the concomitant administration of conventional antibiotics with   which is normally taken as food (spice) will negatively affect its activity. Phytochemical analysis of the extract performed following established methods Results The extract showed good but varying in vitro activities against all the isolates tested. While ampicillin showed synergistic interaction with the plant extract against clinical isolates of E. coli and P. mirabilis, septrin was synergistic against the clinical isolate of E. coli only. Similarly, the activity of the extract against C. albicans isolate was synergistic with ketoconazole and nystatin.

1.3 JUSTIFICATION OF THE STUDY

 Ocimum gratissimum is a well-known plant used in the Indian system of medicine. Folklore medicine claims its use in headache, fever, diarrhoea, pneumonia etc. Research carried out using different in vitro and in vivo techniques of biological evaluation supports most of the claims. This review presents the ethnobotanical, natural product chemistry, pharmacological, clinical and toxicological data of the plant.

1.4 AIM

The aim of this research is to study the thin layer and U.V spectroscopic characteristics of the phytochemical’s presence in Ocimum gratissimum.

1.5 OBJECTIVES OF THE STUDY

       I.            To determine the phytochemical components of Ocimum gratissimum

    II.            To determine the thin layer properties of Ocimum gratissimum

 III.            To determine U.V characteristics of chemicals from Ocimum gratissimum.

 



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