Tulsi: An important medicinal plant

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Tulsi: An important medicinal plant

1.1 General introduction:

History of many drugs can be traced back to their natural origin. At the very dawn of civilization, coexistence of disease with the emergence of life in the earth compelled the primitive man to search a cure from his surroundings. Since then plant was being used as remedy for many diverse disease ranging from simple skin infection to such formidable foes as heart diseases and cancers. The medicinal use of some plants by Indo-Aryans was noted in the Rig Veda (4500-1600BC) and the medicinal value of many plant constituents used by Egyptians was recorded in Papyrus Abus (1500BC).The great physician Hippocrates, who is called ‘the father of medicinal science’, also used plants for treatment of many diseases. In this respect medicine can be considered as an ancient art consisting of plant materials.1

Starting from the stone-age, the use of plants as traditional medicine has increased with the age of civilization. Herbal medicines are still in use in many societies. Although most of the modern medicines are simple compounds, but in many cases the drugs have been originated from the nature, more specifically from plant sources. Plants are considered as natural chemical factory, because synthetic process in biological systems particularly in plants is going on by nature’s ordinary condition of temperature and pressure. The laboratory synthesis of anti-malarial drug quinine requires an intensive work extending over half a century but chicona plant can do it everyday without difficulty. Till today, extensive phytochemical analysis of plants yielded diversified chemical compounds as steroids, terpenoids, flavonoids, chalcones, alkaloids, glycosides etc. clinically used plant metabolites such as taxol isolated from Taxes brevifolia, Vineristine and Vinblastine obtained from Vinca rosea Linn. and digitalis derived from Digitalis purpura are only few of the many striking examples of developing life saving drugs from plant sources.2

Once the plant medication was provided to the ancient people in the crude form, it often exhibited many unwanted effects due to the presence of some toxic components beyond the active constituent. Extensive phytochemical investigation and isolation of active component in the pure form thus became necessary to avoid untoward effects and to ensure safe use of herbal medicines, phytochemical studies of medicinal plants got a rapid pace and the presence of many chemical compounds of diversified nature including many new compounds came in light. These plant derived compounds often played an important role in directing laboratory synthesis of many new classes of drug molecules. In some cases, the plant components became the starting material in the synthetic process of industrial production of many drug molecules. For example, the use of sterol diosgenin, isolated from Mexican yam, for laboratory synthesis of oral contraceptive progesterone reduce the cost of progesterone from a value of $80 per gram to $1.75 per gram.3

Sometimes phytochemical analysis for many plants yielded such chemical constituent having no remarkable therapeutic interest. Sometimes the crude drugs containing several constituents were found to be ineffective in case of therapy for which it is used. The phytochemical investigation of periwinkle plant (Vinca rosea), once used traditionally as an anti-diabetic drug, was found to contain alkaloidal constituents having hypoglycemic potency in minute quantities. The dried seeds of plant Amni visanaga was used as diuretic and antispasmodic in renal collie in Eastern Mediterranean countries and in Arabia, but the research carried out by G.V. Anrep and coworkers resulted in the isolation of Khelin, a component having vasodilator effect. Khelin appeared in an anti-anginel drug after subsequent clinical trial. The research on Ranwolfia serpentine, which was traditionally used as an antidote for snake bite, revealed the presence of an antihypertensive agent reserpine. Thus systemic research with medicinal plant might open the door of many unknown therapeutic choices. Serendipity has also much impact in discovery of drug leads from natural origin.4

From the above discussion it is evident that the search of plant constituents having therapeutic interest requires bioactivity studies with the crude extracts prior to phytochemical investigation. Only the bioactive extracts or fraction would be of interest for next phytochemical analysis. Without prior biological studies, the phytochemical analysis alone can provide the chemical composition of the plant which may or may not have therapeutic interest. Thus in conducting research on medicinal plants, bioactivity guided phytochemical analysis might be a rational approach.5

1.2 Plant Based Drugs and Medicines:

Today there are at least 120 distinct chemical substances derived from plants that are considered as important active compounds for preparation of drugs currently in use in one or more countries in the world. These chemical substances are shown in the table below. Several of the drugs sold today are simple synthetic modifications or copies of the naturally obtained substances. The original plant substance/chemical name is shown under the “Drug” column rather than the finished patented drug name. For example, many years ago a plant chemical was discovered in a tropical plant, Cephaelis ipecacuanha, and the chemical was named emetine.6 a drug was developed from this plant chemical called Ipecac which was used for many years to induce vomiting mostly if someone accidentally swallowed a poisonous or harmful substance. Ipecac can still be found in pharmacies in many third world countries but has been mostly replaced by other drugs in the United States. Another example of this is the plant chemical named taxol shown in the drug column below. The name taxol is the name of the plant chemical originally discovered in the plant. A pharmaceutical company copied this chemical and patented a drug named Paclitaxe which is used in various types of tumors today in the U.S. and many other countries.7-8

The 120 substances given in table-1 are sold as drugs worldwide but not in all countries. Some European countries regulate herbal substances and products differently than in the United States. Many European countries, including Germany, regulate herbal products as drugs and pharmaceutical companies prepare plant based drugs simply by extracting out the active chemicals from the plants.9

Some of the drug/chemicals shown in table-1 are still sold as plant based drugs requiring the processing of the actual plant material. Others have been chemically copied or synthesized by laboratories and no plant materials are used in the manufacture of the drug. A good example of this is the plant chemical quinine, which was discovered in a rainforest tree (Cinchona ledgeriana) over 100 years ago. For many years the quinine chemical was extracted from the bark of this tree and processed into pills to treat malaria. Then a scientist was able to synthesize or copy this plant alkaloid into a chemical drug without using the original tree bark for manufacturing the drug. Today, all quinine drugs sold are manufactured chemically without the use of any tree bark. However, another chemical in the tree called quinidine which was found to be useful for various heart conditions couldn’t be completely copied in the laboratory and the tree bark is still harvested and used to extract this plant chemical from it. Quinidine extracted from the bark is still used today to produce quinidine-based drugs.10

Table-1: List of drugs or chemicals of plant origin having clinical use.

Drug/Chemical Action/Clinical Use Plant Source
Acetyldigoxin Cardiotonic Digitalis lanata
Adoniside Cardiotonic Adonis vernalis
Aescin Anti-inflammatory Aesculus hippocastanum
Aesculetin Anti-dysentery Frazinus rhychophylla
Agrimophol Anthelmintic Agrimonia supatoria
Ajmalicine Circulatory Disorders Rauvolfia sepentina
Allantoin Vulnerary Several plants
Allyl isothiocyanate Rubefacient Brassica nigra
Anabesine Skeletal muscle relaxant Anabasis sphylla
Andrographolide Baccillary dysentery Andrographis paniculata
Anisodamine Anticholinergic Anisodus tanguticus
Anisodine Anticholinergic Anisodus tanguticus
Arecoline Anthelmintic Areca catechu
Asiaticoside Vulnerary Centella asiatica
Atropine Anticholinergic Atropa belladonna
Benzyl benzoate Scabicide Several plants
Berberine Bacillary dysentery Berberis vulgaris
Bergenin Antitussive Ardisia japonica
Betulinic acid Anticancerous Betula alba
Borneol Antipyretic, analgesic, anti-inflammatory Several plants
Bromelain Anti-inflammatory, proteolytic Ananas comosus
Caffeine CNS stimulant Camellia sinensis
Camphor Rubefacient Cinnamomum camphora
Camptothecin Anticancerous Camptotheca acuminata
(+)-Catechin Haemostatic Potentilla fragarioides
Drug/Chemical Action/Clinical Use Plant Source
Chymopapain Proteolytic, mucolytic Carica papaya
Cissampeline Skeletal muscle relaxant Cissampelos pareira
Cocaine Local anaesthetic Erythroxylum coca
Codeine Analgesic, antitussive Papaver somniferum
Colchiceine amide Antitumor agent Colchicum autumnale
Colchicine Antitumor agent, anti-gout Colchicum autumnale
Convallatoxin Cardiotonic Convallaria majalis
Curcumin Choleretic Curcuma longa
Cynarin Choleretic Cynara scolymus
Danthron Laxative Cassia species
Demecolcine Antitumor agent Colchicum autumnale
Deserpidine Antihypertensive, tranquillizer Rauvolfia canescens
Deslanoside Cardiotonic Digitalis lanata
L-Dopa Anti-parkinsonism Mucuna sp
Digitalin Cardiotonic Digitalis purpurea
Digitoxin Cardiotonic Digitalis purpurea
Digoxin Cardiotonic Digitalis purpurea
Emetine Amoebicide, emetic Cephaelis ipecacuanha
Ephedrine Sympathomimetic, antihistamine Ephedra sinica
Etoposide Antitumor agent Podophyllum peltatum
Galanthamine Cholinesterase inhibitor Lycoris squamigera
Gitalin Cardiotonic Digitalis purpurea
Glaucarubin Amoebicide Simarouba glauca
Glaucine Antitussive Glaucium flavum
Glasiovine Antidepressant Octea glaziovii
Glycyrrhizin Sweetener, Addison’s disease Glycyrrhiza glabra
Gossypol Male contraceptive Gossypium species
Drug/Chemical Action/Clinical Use Plant Source
Hemsleyadin Bacillary dysentery Hemsleya amabilis
Hesperidin Capillary fragility Citrus species
Hydrastine Hemostatic, astringent Hydrastis Canadensis
Hyoscyamine Anticholinergic Hyoscyamus niger
Irinotecan Anticancer, antitumor agent Camptotheca acuminata
Kaibic acud Ascaricide Digenea simplex
Kawain Tranquillizer Piper methysticum
Kheltin Bronchodilator Ammi visage
Lanatosides A, B, C Cardiotonic Digitalis lanata
Lapachol Anticancer, antitumor Tabebuia sp.
a-Lobeline Smoking deterrant, respiratory stimulant Lobelia inflate
Menthol Rubefacient Mentha species
Methyl salicylate Rubefacient Gaultheria procumbens
Monocrotaline Antitumor agent (topical) Crotalaria sessiliflora
Morphine Analgesic Papaver somniferum
Neoandrographolide Dysentery Andrographis paniculata
Nicotine Insecticide Nicotiana tabacum
Nordihydroguaiaretic acid Antioxidant Larrea divaricata
Noscapine Antitussive Papaver somniferum
Ouabain Cardiotonic Strophanthus gratus
Pachycarpine Oxytocic Sophora pschycarpa
Palmatine Antipyretic, detoxicant Coptis japonica
Papain Proteolytic, mucolytic Carica papaya
Papavarine Smooth muscle relaxant Papaver somniferum
Phyllodulcin Sweetner Hydrangea macrophylla
Physostigmine Cholinesterase Inhibitor Physostigma venenosum
Picrotoxin Analeptic Anamirta cocculus
Drug/Chemical Action/Clinical Use Plant Source
Pilocarpine Parasympathomimetic Pilocarpus jaborandi
Pinitol Expectorant Several plants
Podophyllotoxin Antitumor anticancer agent Podophyllum peltatum
Protoveratrines A, B Antihypertensives Veratrum album
Pseudoephredrine* Sympathomimetic Ephedra sinica
Pseudoephedrine, nor- Sympathomimetic Ephedra sinica
Quinidine Antiarrhythmic Cinchona ledgeriana
Quinine Antimalarial, antipyretic Cinchona ledgeriana
Qulsqualic acid Anthelmintic Quisqualis indica
Rescinnamine Antihypertensive, tranquillizer Rauvolfia serpentine
Reserpine Antihypertensive, tranquillizer Rauvolfia serpentine
Rhomitoxin Antihypertensive, tranquillizer Rhododendron molle
Rorifone Antitussive Rorippa indica
Rotenone Piscicide, Insecticide Lonchocarpus nicou
Rotundine Analagesic, sedative, traquillizer Stephania sinica
Rutin Capillary fragility Citrus species
Salicin Analgesic Salix alba
Sanguinarine Dental plaque inhibitor Sanguinaria Canadensis
Santonin Ascaricide Artemisia maritma
Scillarin A Cardiotonic Urginea maritime
Scopolamine Sedative Datura species
Sennosides A, B Laxative Cassia species
Silymarin Antihepatotoxic Silybum marianum
Sparteine Oxytocic Cytisus scoparius
Drug/Chemical Action/Clinical Use Plant Source
Stevioside Sweetner Stevia rebaudiana
Strychnine CNS stimulant Strychnos nux-vomica
Taxol Antitumor agent Taxus brevifolia
Teniposide Antitumor agent Podophyllum peltatum

(THC)Antiemetic, decrease occular tensionCannabis sativaTetrahydropalmatineAnalgesic, sedative, traquillizerCorydalis ambiguaTetrandrineAntihypertensiveStephania tetrandraTheobromineDiuretic, vasodilatorTheobroma cacaoTheophyllineDiuretic, bronchodilatorTheobroma cacao and othersThymolAntifungal (topical)Thymus vulgarisTopotecanAntitumor, anticancer agentCamptotheca acuminataTrichosanthinAbortifacientTrichosanthes kirilowiiTubocurarineSkeletal muscle relaxantChondodendron tomentosumValapotriatesSedativeValeriana officinalisVasicineCerebral stimulantVinca minorVinblastineAntitumor, Antileukemic agentCatharanthus roseusVincristineAntitumor, Antileukemic agentCatharanthus roseusYohimbineAphrodisiacPausinystalia yohimbeYuanhuacineAbortifacientDaphne genkwaYuanhuadineAbortifacientDaphne genkwa

1.3 Role of Plant on Cancer and Aids Research:

The National Cancer Institute (NCI) has several ongoing collaborative programs which screen plants for the possibility of new drugs and active plant chemicals for cancer and AIDS/HIV.

As well over 50 percent of the estimated 250,000 plant species found on earth come from tropical forests, NCI concentrates on these regions. Plants have been collected from the African countries of Cameroon, the Central African Republic, Gabon, Ghana, Madagascar, and Tanzania. Collections are now concentrated in Madagascar (one of the most rapidly disappearing rainforest regions in the world), and collaborative programs have been established in South Africa and Zimbabwe.

In Central and South America, samples have been collected from Belize, Bolivia, Colombia, the Dominican Republic, Ecuador, Guatemala, Guyana, Honduras, Martinique, Paraguay, Peru, and Puerto Rico. The NCI has established collaborative programs in Brazil, Costa Rica, Mexico, and Panama. Southeast Asian collections have been performed in Bangladesh, Indonesia, Laos, Malaysia, Nepal, Pakistan, Papua New Guinea, the Philippines, Taiwan, Thailand, and Vietnam. Collaborative programs have been established in Bangladesh, China, Korea, and Pakistan. In each country, NCI contractors work in close collaboration with local botanical institutions.14

Thus far seven plant-derived anticancer drugs have received Food and Drug Administration (FDA) approval for commercial production: 15

• Taxol/paclitaxel

A chemical discovered in the Pacific Yew tree (Taxus brevifolia) is now the first drug of choice in several tumorous cancers including Breast Cancer.


A chemical discovered in the Madagascar Periwinkle in the 1950s. Vinblastine is the first drug of choice in many forms of leukemia and since the 1950’s it has increased the survival rate of childhood leukemias by 80%.

• Vincristine

Another antileukemic drug discovered in the Madagascar periwinkle.

• Topotecan

It has been approved by the FDA for the treatment of ovarian and small cell lung cancers. It is currently in clinical trials, either alone or in combination with other anticancer drugs, for several types of cancer. Topotecan is an analog (a synthesized chemical) of a plant alkaloid discovered in the Chinese tree species, Camptotheca acuminata.

• Irinotecan

Another chemical analog which has been developed from yet another plant alkaloid discovered in the same tree Camptotheca acuminata. It has been approved by the FDA for the treatment of metastatic colorectal cancer. It is currently in clinical trials for a variety of other cancers.

• Etoposide

It is a semi synthetic derivative of a plant chemical epipodophyllotoxin discovered in the May apple plant family (Podophyllum peltatum).

• Teniposide

It is another semi synthetic derivative of a plant chemical discovered in the May apple plant family (Podophyllum peltatum).

Since 1986, over 40,000 plant samples have been screened, but thus far only five chemicals showing significant activity against AIDS have been isolated. Three are currently in preclinical development. Before being considered for clinical trials in humans, these agents must show tolerable levels of toxicity in several animal models. For AIDS, three agents are presently in preclinical or early clinical development. The following are plants and chemicals which are still under research for cancer and AIDS/HIV:

• (+)-Calanolide A and (-)-Calanolide B (costatolide) are isolated from Calophyllum lanigerum and Calophyllum teysmanii, respectively, trees found in Sarawak, Malaysia. Both these agents are licensed to Medichem, Inc., Chicago, which is developing them in collaboration with the Sarawak State Government through a joint company, Sarawak Medichem Pharmaceuticals, Inc.(+)-Calanolide A is currently in early clinical trials in the United States.

• Conocurovone, isolated from the shrub species, Conospermum incurvum (saltbush), found in Western Australia, has been licensed for development to AMRAD, a company based in Victoria, Australia.

• Michellamine B, from the leaves of Ancistrocladus korupensis, a vine found in the Korup rainforest region of southwest Cameroon, has undergone extensive preclinical study, but is considered too toxic for advancement to clinical trials.

• Prostratin, isolated from the wood of Homolanthus nutans, a tree found in Western Samoa, has been placed on low priority, largely due to its association with a class of compounds shown to be tumor promoters.

• A tree native to China, Camptotheca acuminate, is the source of four promising anticancer drugs, two of which have been approved by the FDA and are described above. The other two chemicals still under research include:

• 9AC (9-aminocamptothecin): Currently in clinical trials for several types of cancer, including ovarian and stomach cancers and T-cell lymphoma.

• Camptothecin: While no clinical trials are being performed in the United States, trials are ongoing in China.

• Homoharringtonine from the Chinese tree, Cephalotaxus harringtonia are in early clinical trials.

• Perillyl alcohol, and flavopiridol, a totally synthetic compound based on a flavone isolated from Dysoxylum binectiferum are in early clinical trials.16-17

1.4 Bioactivity guided phytochemical investigation of the plants:

The use of plant products is increasing in many segment of the population. At present thousands of plant metabolics are being successively used in the treatment of variety of diseases. According to estimation 40% of the world’s populations rely upon plants for their medication. The use of medicinal plants is increasing in many developed countries where 35% of drugs involve natural products. Since Bangladesh has a vast resource of medicinal plants, the present study might be a significant way of making the best use of these natural resources. Majority of our population, who are impoverished, have to rely upon indigenous system of medication because of their inability to meet the cost of modern medicine. Thus in order to strengthen the existing health care system, biological activity directed chemical analysis of indigenous plant Ocimum sanctum is the primary objective of bioactivity directed phytochemical investigation of plant products. Moreover the standardization of herbal medicines has made them popular in many developed countries. So the economic impact of the present study may be reflected through export standard, high quality herbal drugs, which would increase our natural reserve. Thus the rationality of the present study lies in meeting the challenge of developing herbal medicine for the coming twenty first century which needs a systematic research on indigenous medicinal plants for the welfare of the humanity.

1.5 Objective:

Main objectives of the proposed research program are:

    1. To extract and isolate biologically active compounds from the leaves of O. sanctum using various solvents such as methanol, chloroform, ethyl acetate, n-hexane and butanol and to identify the structure of the isolated compounds using various analytical and instrumental techniques including UV, IR, 1H-NMR, 13C-NMR and mass spectroscopy.
    1. To evaluate the medicinal value of the Isolated Compounds obtained from the extracts of the leaves of O. sanctum by means of several studies related to antibacterial, antifungal.
    1. To identify and suggest the possible area of the utilization of the isolated compounds from the extract of seeds in the cure of various diseases.

1.6 Literature Survey:

1.7 General Information:

Kingdom : Plantae
Subkingdom : Tracheobionta
Superdivision : Spermatophyta
Division : Magnoliophyta
Class : Magnoliopsida
Order : Lamiales
Family : Lamiaceae
Genus : Ocimum
Species : O. sanctum
Indian Name: Tulsi
Botanical or Latin Name : Ocimum sanctum
English name : Sacred Basil / Holy Basil

1.8 Origin of Ocimum sanctum:

Holy basil is native to tropical Asia but has been dispersed by humans so that it now grows in many tropical parts of the world. It is a sacred plant in Hindu religion, and has been cultivated in India in courtyards or temples, and in pots in homes, for about 3000 years.

Holy basil has been cultivated in India for thousands of years.

1.9 History:

The history of the plant in South Asia is closely linked with folklore and mythology. It represents Vishnupriya or Beloved of Vishnu, since it is believed to be the embodiment of the goddess Lakshmi, the spouse of Vishnu. What is apparent is that it has been valued and cultivated since ancient times in India as an intimate link between the household and the spiritual world.

The Aryans, who structured the forms of Hinduism, were nature-worshippers and their poetry and imagery were rich with the evocation of nature. Perhaps they were drawn to holy basil because of its fragrance and delicacy. It may also have been already well-entrenched in the myths of the indigenous people and from there absorbed into Hinduism.

Holy basil is mentioned in the Rig Veda, written in about 1500 BC, and its holiness is celebrated in the Puranas. It is highly regarded in the Ayurvedic system of medicine and is noted in medical treatises such the Charaka Samhita written between the 2nd century BC to the 2nd century AD.

1.10 Description of OCIMUM SANCTUM:

Tulsi plant, is a shrub reaching a height of 0.5 to 1.5 m. The leaves are 2-4 cms in length. There are several varieties of the plant. However, commonly used one is with dark leaves. The inflorescence is a long spike with tiny purple flowers. There are two varieties: a red- and a green one. Red holy basil has a stronger smell. It is cultivated in gardens throughout Bengal, /East Nepal and Deccan Peninsula; it is said to be a common wild plant in Western India.

1.11 Characteristics of Constituents:

The leaves contain an essential oil which has been studied with gas chromatography. The oil contains eugenol, eugenal, carvacrol, methylchavicol, limatrol and caryophylline. The seeds contain an oil composed of fatty acids and sitosterol. The mucilage is compared of sugars – xylose and polysaccharides.

Tulasi as an Ayurvedic medicine

Tulasi, as used in Ayurveda.

Tulasi’s extracts are used in ayurvedic remedies for common colds, headaches, stomach disorders, inflammation, heart disease, various forms of poisoning, and malaria. Traditionally, tulasi is taken in many forms: as an herbal tea, dried powder, fresh leaf, or mixed with ghee. Essential oil extracted from Karpoora Tulsi is mostly used for medicinal purposes and in herbal toiletry. For centuries, the dried leaves of Tulasi have been mixed with stored grains to repel insects.

Recent studies suggest that Tulasi may be a COX-2 inhibitor, like many modern painkillers, due to its significant amount of Eugenol (1-hydroxy-2-methoxy-4-allylbenzene).<href=”#_note-4″ title=””>[5]<href=”#_note-5″ title=””>[6] Studies have also shown Tulsi to be effective for diabetes, by reducing blood glucose levels.<href=”#_note-6″ title=””>[7] The same study showed significant reduction in total cholesterol levels with Tulsi. Another study showed that Tulsi’s beneficial effect on blood glucose levels is due to its antioxidant properties.<href=”#_note-7″ title=””>[8]

Tulasi also shows some promise for protection from radiation poisoning<href=”#_note-8″ title=””>[9] and cataracts.<href=”#_note-9″ title=””>[10] Some Vaishnavites do not use Tulasi for medicine, though, out of reverence. However, the use of Tulsi for purification and as a medicine is widespread throughout India. Many Hindus — along with the ancient tradition of Ayurveda — believe that the healing properties of sacred herbs such as Tulsi were given by the Lord himself, and can be used as a medicine out of reverence.

1.12 Parts used and where grown

Holy basil is native to the Indian subcontinent and other parts of tropical Asia. The leaf and seed oil are used therapeutically.

Holy basil has been used in connection with the following conditions (refer to the individual health concern for complete information):

Science Ratings Health Concerns

Type 2 diabetes Poison oak and poison ivy dermatitisReliable and relatively consistent scientific data showing a substantial health benefit.

Contradictory, insufficient, or preliminary studies suggesting a health benefit or minimal health benefit.

For a herb, supported by traditional use but minimal or no scientific evidence. For a supplement, little scientific support and/or minimal health benefit.

1.13 Varieties of Holy Basil-Tulsi:

Krishna Tulsi Rama Tulsi Vana Tulsi

1.14 Active compounds:

The essential oil from some populations of holy basil contains high levels of eugenol. This compound has anti-inflammatory activity, can kill bacteria and deters insects. The presence of this compound in the plant could explain why it could be used to treat pain, kill germs and provide people with some protection from being bitten by insects.

Another compound called rosmarinic acid has anti-inflammatory and antioxidant activity and these activities could contribute too many of the medicinal properties of holy basil. The plant also contains ursolic acid a compound that has been shown to provide some protection to enzymes in the liver that deal with the breakdown of fat in our diet. This is important as patients with diabetes often have high levels of cholesterol in their blood. The levels have been reported to decrease after taking holy basil.

1.15 Medicinal values of the Tulsi:

The tulsi or holy basil is an important symbol in the Hindu religious tradition and is worshipped in the morning and evening by Hindus at large. The holy basil is also a herbal remedy for a lot of common ailments. Here’re top fifteen medicinal uses of tulsi.

1. Healing Power: The tulsi plant has many medicinal properties. The leaves are a nerve tonic and also sharpen memory. They promote the removal of the catarrhal matter and phlegm from the bronchial tube. The leaves strengthen the stomach and induce copious perspiration. The seed of the plant are mucilaginous.

2. Fever & Common Cold: The leaves of basil are specific for many fevers. During the rainy season, when malaria and dengue fever are widely prevalent, tender leaves, boiled with tea, act as preventive against theses diseases.

In case of acute fevers, a decoction of the leaves boiled with powdered cardamom in half a liter of water and mixed with sugar and milk brings down the temperature. The juice of tulsi leaves can be used to bring down fever. Extract of tulsi leaves in fresh water should be given every 2 to 3 hours. In between one can keep giving sips of cold water. In children, it is every effective in bringing down the temperature.

3. Coughs: Tulsi is an important constituent of many Ayurvedic cough syrups and expectorants. It helps to mobilize mucus in bronchitis and asthma. Chewing tulsi leaves relieves cold and flu.

4. Sore Throat: Water boiled with basil leaves can be taken as drink in case of sore throat. This water can also be used as a gargle.

5. Respiratory Disorder: The herb is useful in the treatment of respiratory system disorder. A decoction of the leaves, with honey and ginger is an effective remedy for bronchitis, asthma, influenza, cough and cold. A decoction of the leaves, cloves and common salt also gives immediate relief in case of influenza. They should be boiled in half a liter of water till only half the water is left and add then taken.

6. Kidney Stone: Basil has strengthening effect on the kidney. In case of renal stone the juice of basil leaves and honey, if taken regularly for 6 months it will expel them via the urinary tract.

7. Heart Disorder: Basil has a beneficial effect in cardiac disease and the weakness resulting from them. It reduces the level of blood cholesterol.

8. Children’s Ailments: Common pediatric problems like cough cold, fever, diarrhea and vomiting respond favorably to the juice of basil leaves. If pustules of chicken pox delay their appearance, basil leaves taken with saffron will hasten them.

9. Stress: Basil leaves are regarded as an ‘adaptogen’ or anti-stress agent. Recent studies have shown that the leaves afford significant protection against stress. Even healthy persons can chew 12 leaves of basil, twice a day, to prevent stress. It purifies blood and helps prevent several common elements.

10. Mouth Infections: The leaves are quit effective for the ulcer and infections in the mouth. A few leaves chewed will cure these conditions.

11. Insect Bites: The herb is a prophylactic or preventive and curative for insect stings or bites. A teaspoonful of the juice of the leaves is taken and is repeated after a few hours. Fresh juice must also be applied to the affected parts. A paste of fresh roots is also effective in case of bites of insects and leeches.

12. Skin Disorders: Applied locally, basil juice is beneficial in the treatment of ringworm and other skin diseases. It has also been tried successfully by some naturopaths in the treatment of leucoderma.

13. Teeth Disorder: The herb is useful in teeth disorders. Its leaves, dried in the sun and powdered, can be used for brushing teeth. It can also be mixed with mustered oil to make a paste and used as toothpaste. This is very good for maintaining dental health, counteracting bad breath and for massaging the gums. It is also useful in pyorrhea and other teeth disorders.

14. Headaches: Basil makes a good medicine for headache. A decoction of the leaves can be given for this disorder. Pounded leaves mixed with sandalwood paste can also be applied on the forehead for getting relief from heat, headache, and for providing coolness in general.

15. Eye Disorders: Basil juice is an effective remedy for sore eyes and night-blindness, which is generally caused by deficiency of vitamin A. Two drops of black basil juice are put into the eyes daily at bedtime.

1.16 Safety

Holy basil has a long history of safe use in India. Application to the skin can cause reactions in sensitive people.

1.17 Chemical investigation of the genus Ocimum sanctum(Literature review) :


3.1 Investigation On Ocimum Sanctum

3.2 Collection Of Plants

The plant Ocimum Sanctum was collected from Gazipur area and was identified from the department of Botany, Dhaka University. The collected fresh plants were cleaned thoroughly. The plant materials (leaves and stems) were separated from its roots and dried under mild sunlight and then at 40 oC in an oven. Afterwards the plants were powdered in a grinding machine (~200 smeshes). The powder was used throughout the investigation.

3.3 Phytochemical Screening Of The Plants

Chemical tests were carried out on the aqueous extract and on the powdered specimens using standard procedures to identify the constituents .

3.4 Qualitative Determination

Test For Tannins

The dried powder samples (~0.5 g) was boiled in water (20 ml) in a test tube and then filtered. A few drops of ferric chloride solution (0.1%w/v) were added and brownish green or a blue-black coloration was observed.

Test For Phlobatannins

Deposition of a red precipitate (on boiling) extract of each plant sample was boiled with aqueous hydrochloric acid (1% w/v) indicated the presence of phlobatannins.

Test For Saponin

The powdered sample (~2g) was boiled in distilled water (20 ml) in a water bath and filtered. The filtrate (10 ml) was mixed with distilled water (5 ml) and shaken vigorously for a stable persistent froth. The frothing was mixed with olive oil (3 drops) and shaken vigorously, and then the formation of emulsion was observed.

Test For Flavonoids

Three methods were used to determine the presence of flavonoids in the plant sample-

a) Dilute ammonia solution (5 ml) was added to a portion of the aqueous filtrate of each plant extract followed by addition of concentratedH2SO4.

b) Few drops of aluminium ion (Al3+) solution (1%) were added to a portion of each filtrate. A yellow colouration each observed indicating the prsence of flavonoids.

c) A portion of the powdered plant sample each case was heated with ethyl acetate (10 ml) over a steam bath for 3 min. The mixture was filtered and the filtrate (4ml) was shaken with dilute ammonia solution (1 ml). A yellow colouration was observed indicating a positive test for flavonoids.

Test For Steroids

Acetic anhydride (2 ml) was added to ethanolic extract (0.5 g) of each sample with H2SO4 acid (2 ml). The colour changed from violet to blue or green in some samples indicating the presence of steroids.

Test For Terpenoids (Salkowski Test)

Each extract (5 ml) was mixed in chloroform (2 ml), and concentrated H2SO4 acid (3 ml) was carefully added to form a layer. A reddish brown colouration at the interface was formed to show positive test for the presence of terpenoids.

Test For Cardiac Glycosides (Keller-Killani Test)

Each extracted (5 ml) was treated with glacial acetic acid (2 ml) containing ferric chloride solution (1 drop). This was underlayed with concentrated H2SO4 acid (1 ml). A brown ring at the interface indicates a deoxysugar characteristic of cardinolides. A violet ring may appear bellow the brown ring, while in the acetic acid layer, greenish ring may from just gradually throughout thin layer.

Table-3.1: Qualitative analysis of the phytochemical of the medicinal plants

Plants Alkaloid Tannin Saponin Flavonoid Steroid Terpenoid Cardiac glycoside
Ocimum Sanctum + + + + + + +

3.5 Quantitative Determination Of The Chemical Constituents

Alkaloid Determination 20

Each sample (0.5 g) was taken into a conical flask (100 ml) and acetic acid (20 ml in 10%) ethanol was added. It was covered and allowed to stand for 12 hours. This was filtered and the extract was concentrated on water bath to one-quarter of the original volume. Concentrated ammonium hydroxide was added drop wise to the extract until the precipitation was complete. The whole solution was allowed to stand and the precipitate was collected and washed with dilute ammonium hydroxide solution and then filtered. The residue is the alkaloid, which was dried and weighed.

Flavonoid Determination 22

The plant sample (1 g) was extracted repeatedly with aqueous methanol (20 ml in 80%) (CH3OH) at room temperature. The whole solution was filtered through whatman filter paper No 42 (125 mm). The filtrate was later transferred into a crucible and evaporated into dryness over a water bath ansd weighed to a constant weight.

Saponin Determination 23

Each of the powder samples (5 g) were put into a conical flask and 20% aqueous ethanol (25 ml) was added. The samples were heated over a hot water bath for 4 hours with continuous stirring at about55oC. The mixture was filtered and the residue re-extracted with 20% aqueous ethanol (50 ml). The combined extracts were reduced to 10 ml over water bath at about 90oC. The concentrated extract (was transferred into a separatory funnel and diethyl ether (5 ml) was added and shaken vigorously. The aqueous layer was recovered while the whether layer discarded. The purification process was repeated. n-butanol (15 ml) wad added and n-butanol extracts were washed twice with aqueous 5% NaCl (3 ml of) solution). The remaining solution was heated on a water bath. After evaporation the samples were dried in an oven to a constant weight; the saponin content calculated as percentage. .

Table-3.2: Amount (% on dry powder basis) of crude alkaloid, flavonoid and saponin on the medicinal plants investigated.

Plants Alkaloids (%) Flavonoids (%) Saponin (%)
Ocimum Sanctum 0.8 14.06 8.5

3.6 Extraction And Isolation Of Compounds From Ocimum Sanctum

Plant powder was taken in a few precleaned cloth thimbles. The thimbles containing the powder were placed in a Soxhlet apparatus.

The plant powder extracted separately and exhaustively in Soxhlet apparatus first with Petroleum ether (boiling point 40-60 oC and 60-80 oC) followed by hexane. All the extracts were filtered individually; the filtrate was combined together and then concentrated in a “Buchi Rotavapor” under reduced pressure. This was shown in scheme-1


Powder (80g)

Extraction with pet-ether (bp.40-60oC)

Petroleum ether exract Residue

Evaporated to dryness Extracted with hexane

Gummy mass Hexane extract Residue

Evaporated to dryness

Dry mass of hexane extract (18 g)

Scheme 1: Extraction scheme of Ocimum Sanctum

3.7 Investigation Of Hexane Extract

3.8 Thin Layer Chromatography (TLC) Study

TLC analysis of the hexane extract showed the presence of one spot (Petroleum ether: Dichloromethane = 90: 20) having light brown under iodine vapor. Furthermore, TLC study showed the presence of two spots on spraying with vanillin sulfuric acid followed by heating in oven for 15 minutes. Again three spots (Dichloromethane: Ethyl acetate = 80: 20) were visible on spraying vanillin sulfuric acid followed by heating in an oven for 15 minutes. Among these three spots, two were pink and one was violet. The presence of the pink colored spot was thought to be due to the presence of steroidal or fatty acid material or both of them. It gave also positive test for steroid23.

3.9 Fractionated By Vaccum Liquid Chromatography (VLC)

The hexane extract was concentrated to dry mass about 18g by a ‘Buchi Rotavapor’. The concentrated extract was mixed with TLC grade silica gel (60 GF254). The mixture was made as powder for using in column. Then the sample was placed on the top of the bed of silica gel packed column (VLC). The column was then eluted with petroleum ether followed by mixtures of pet-ether and dichloromethane of increasing polarity. The elutes were collected in test tube with an about of about 20mL in each. Solvent system used as mobile phase in the analysis of hexane extract was listed in Table.

Table- Number of fractions collected in test tubes from VLC of hexane extract by using different solvent system

Solvent system Amounts

of solvents

(mL)Number of test tubesPet- ether (%)DCM

(%)Ethyl acetate


3.10 Screening Of The Fractions

TLC monitored each of the fractions and the fractions of similar behaviors were combined together and marked as F1, F2, F3, F4, F5, F6, F7, . This is given in Table

Table-: Screening of the fractions by the similar TLC pattern

No of test tubes Fractions Solvent system for TLC TLC pattern
1-12 F1 90:10 (PE:DCM) One spot
17-19 F2 90:10 (PE:DCM) One spot with tailing
32-35 F3 70:30 (PE:DCM) Two spots with tailing
62-64 F4 20:80 (DCM:PE) One spot
92-100 F5 20:80 (PE:DCM) One spot with tailing
101-103 F6 20:80 (PE:DCM) no spots
119-121 F7 20:80 (EA:DCM) two spots with tailing

3.11 Attempt Of Purification And Characterization of the Fractions