Chapter I

General Introduction

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1. Introduction

The relationship existing between plants and humans is as old as mankind, dating back to

the origin of human civilization. Humans have relied on plants for food, clothing, shelter, fuel

and medicine (Newman et al., 2000). Plants continue to be an integral part of human existence,

making ethnobotany an interesting and important research field (Sumner, 2000; Okujagu et al.,

2006). Gurib-Fakim (2006) has identified plants as forming the basis of a sophisticated

traditional medicine from antiquity to date. Most medicinal plants were used against ailments

such as coughs, colds, parasitic infections and inflammation; probably based on trial and error.

This ancient knowledge was verbally transferred from one generation to another (Gurib-Fakim,

2006). Therapeutic plant use can be a herbal tea, a crude extract, a phytopharmaceutical or herbal

mixture or isolated compounds (Rates, 2001).

Globally, many pharmacological and economically noteworthy medicinal plant species

have had multiple uses (Ajibesin et al., 2008). Medicinal plants are found and frequently used in

China, India, Japan, Pakistan, Thailand and South Africa (Mukhtar et al., 2008). Globally, the

Indian Ayurvedic and Chinese traditional medicines are recognized as the oldest and most

developed respectively (Gurib-Fakim, 2006). Traditional medicine also exists in southern

America and Australia, but these are not as developed as in Asia or Africa. Distinct traditional

medicinal systems are found universally in each geographical region, so that medicinal plant use

is diverse globally (Ajibesin et al., 2008).

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1.1 Medicinal plants: Importance and scope

Herbs are staging a comeback and herbal „renaissance‟ is happening all over the globe.

The herbal products today symbolise safety in contrast to the synthetics that are regarded as

unsafe to human and environment. Although herbs had been priced for their medicinal,

flavouring and aromatic qualities for centuries, the synthetic products of the modern age

surpassed their importance, for a while. However, the blind dependence on synthetics is over and

people are returning to the naturals with hope of safety and security.

It has been estimated that in developed countries such as United States, plant drugs

constitute as much as 25% of the total drugs, while in fast developing countries such as China

and India, the contribution is as much as 80%. Thus, the economic importance of medicinal

plants is much more to developing countries such as India than to rest of the world. These

countries provide two third of the plants used in modern system of medicine and the health care

system of rural population depend on indigenous systems of medicine. Of the 2, 50,000 higher

plant species on earth, more than 80,000 are medicinal. India is one of the world‟s 12

biodiversity centres with the presence of over 45000 different plant species. India‟s diversity is

unmatched due to the presence of 16 different agro-climatic zones, 10 vegetation zones, 25 biotic

provinces and 426 biomes (habitats of specific species). Of these, about 15000-20000 plants

have good medicinal value. However, only 7000-7500 species are used for their medicinal values

by traditional communities. This proportion of medicinal plants is the highest proportion of

plants known for their medical purposes in any country of the world for the existing flora of that

respective country. The drugs are derived either from the whole plant or from different organs,

like leaves, stem, bark, root, flower, seed, etc. Some drugs are prepared from excretory plant

product such as gum, resins and latex. Even the Allopathic system of medicine has adopted a

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number of plant-derived drugs which form an important segment of the modern pharmacopoeia.

Some important chemical intermediates needed for manufacturing the modern drugs are also

obtained from plants (Eg. diosgenin, solasodine, -ionone). Not only that plant-derived drug

offers a stable market worldwide, but also plants continue to be an important source for new

drugs.

The Indian Systems of Medicine are in the list approved by the National Council for

Alternative Medicine (NCAM), USA, although most of these drugs that claim to cure various

ailments are yet to be validated scientifically. According to Parasnis (2004), the full potential of

Ayurveda can be realized only by subjecting the ayurvedic drugs to modern investigation

techniques. Further acceptance of any clinical trial depends on whether it satisfies modern

pharmacological and statistical standards or not. Unfortunately, most people practicing Ayurveda

decline to adopt modern research techniques for evaluation of the formulations used in the

practice. This holds good for other traditional medical practitioners like those of Siddha and

Unani medicinal systems too. Scientific validation will not only popularize these medicines in

India but also render them acceptable, in some form, to people in other parts of the world.

Considering the fact that several diseases do not have an ultimate answer in the conventional

system whether in native regions or throughout the world, an effort to recognize the potential of

alternative and combinational treatment systems validated through universally acceptable

methods could prove to be very beneficial for the human community at large.

1.2 Drug discovery from medicinal plants

Rates (2001) defined a drug as “a pharmacologically active compound, which is a

component of a medicine, irrespective of its natural, biotechnological or synthetic origin”. It is

used in the treatment, cure, prevention or diagnosis of diseases. According to WHO (1998), “a

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medicine is a product prepared according to legal and technical procedures, which is used for the

diagnosis, prevention and treatment of disease and has been scientifically characterized in terms

of its efficacy, safety and quality”. Many methods have been used in obtaining compounds for

drug production, these include isolation from plants and other natural sources like microbes,

synthetic chemistry, combinatorial chemistry and molecular modelling (Balunas and Kinghorn,

2005). There are challenges associated with each method, necessitating the need for one method

to complement the other (Shu, 1998).

Natural product sources such as plants and animals have been utilized as drugs for

centuries, with a deep attachment to the socio-cultural life of early humans (Rates, 2001). The

isolation of morphine from opium (Papaver somniferum) by the German pharmacist Friedrich

Sertϋrner in 1805 marked the beginning of isolation of pure compounds from plants (Kinghorn,

2001). This development led to the isolation of drugs such as cocaine (Erythroxylum coca),

codeine (Papaver somniferum), digitoxin (Digitalis species) and quinine (Cinochona species),

with the majority still in use (Newman et al., 2000; Butler, 2004). Newman et al. (2003)

researched 55 categorized human diseases (such as cancer, microbial and parasitic infections)

and found that 87% of the medications in use were derived from natural products from plants.

There are many challenges encountered in the drug discovery process, some of these include the

high cost involved, which has been estimated to be over US$ 800 million, the long period

of time required for the process (minimum of 10 years), and the low average yield of isolated

compounds from natural products which are mostly insufficient for lead optimization,

development and clinical trials (Reichert, 2003; Dickson and Gagnon, 2004; Balunas and

Kinghorn, 2005). This process of drug discovery is also characterized by a very low chance of

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success as it has been estimated that only one in 5000 lead compounds will successfully advance

through clinical trials for final approval as a drug (Balunas and Kinghorn, 2005).

Despite the challenges and obstacles encountered in drug discovery from plants, a

number of clinically useful isolated bioactive compounds from natural products are currently in

use and many are also being investigated (Newman et al., 2003; Balunas and Kinghorn, 2005).

For this reason, natural products isolated from plants have been predicted to continue to remain

an essential part of the search for novel medicines against human diseases (Newman et al., 2000;

Balunas and Kinghorn, 2005). To achieve significant success, Butler (2004) emphasized the need

for all concerned scientists to develop faster and better techniques for plant collection, crude

extract preparation, bioassay screening, compound(s) isolation and development to keep pace

with other drug discovery efforts. There is a bright future for the discovery and development of

more drugs from plants due to the large number of plant species that have not been

pharmacologically and phytochemically investigated (Sumner, 2000; Gurib-Fakim, 2006). The

use of medicinal plants will continue to play an important role as an effective health aid,

especially for the large number of poor people in the third world and developing countries.

Plant-based pharmaceutical products with proven medical efficacy, phytomedicines are

generally prescription drug products with formal market authorization involving detailed

toxicological and clinical trials.

1.3 Traditional and affordable health care

South Asia is home to many rich, traditional systems of medicine. Ayurvedic methods

date back to 5000 B.C. Along with the Unani, Siddha and Tibetan systems, they remain an

important source of everyday health and livelihood for tens of millions of people. Ayurveda, the

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oldest medical system in Indian sub-continent, has alone reported approximately 2000 medicinal

plant species, followed by Siddha and Unani. The Charak Samhita, an age-old written document

on herbal therapy, reports on the production of 340 herbal drugs and their indigenous uses

(Prajapati et al., 2003). Medicinal and aromatic plants (MAPs), including trees, shrubs, grasses

and vines, are a central resource for these traditional health systems, as well as for

pharmaceutical (or allopathic) medicines. There are more than 8,000 plant species in South Asia

with known medicinal uses.

Medicinal plants are an accessible, affordable and culturally appropriate source of

primary health care for more than 80% of Asia's population (WHO). Marginalized, rural and

indigenous people, who cannot afford or access formal health care systems, are especially

dependent on these culturally familiar, technically simple, financially affordable and generally

effective traditional medicines. As such, there is widespread interest in promoting traditional

health systems to meet primary health care needs. This is especially true in South Asia, as prices

of modern medicines spiral and governments find it increasingly difficult to meet the cost of

pharmaceutical-based health care. Throughout the region, there is strong and sustained public

support for the protection and promotion of the cultural and spiritual values of traditional

medicines.

1.4 Widespread and rising demand for medicinal plants

WHO has estimated the present demand for medicinal plants as approximately US $14

billion per year. The demand for medicinal plant-based raw materials is growing at the rate of 15

to 25% annually, and according to an estimate of WHO, the demand for medicinal plants is

likely to increase more than US $5 trillion in 2050. In India, the medicinal plant-related trade is

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estimated to be approximately US $1 billion per year. According to an estimate, the quantity of

export of Ayurvedic products produced in India is tripled (Sharma, 2004).

The projected escalating demand of medicinal plants has led to the over-harvesting of

many plants from wild, which subsequently results in the loss of their existing populations. For

example, the large quantity of Himalayan yew (Taxus baccata) has been gathered from the wild

since its extract, taxol, was established in the treatment of ovarian cancer. Aconitum

heterophyllum, Nardostachys grandiflora, Dactylorhiza hatagirea, Polygonatum verticillatum,

Gloriosa superba, Arnebia benthamii and Megacarpoea polyandra are other examples of Indian

medicinal plant species which have been overexploited for therapeutic uses and have

subsequently been placed today in rare and endangered categories. Many medicinal plant species

are used in curing more than one disease (Kala et al., 2004; Kala et al., 2005), and as a result,

these species are under pressure due to over collection from wild. For example, Hemidesmus

indicus is used to cure 34 types of diseases; Aegle marmelos 31, Phyllanthus emblica 29, and

Gloriosa superba 28. Over-exploitation and continuous depletion of medicinal plants have not

only affected their supply and loss of genetic diversity, but have seriously affected the

livelihoods of indigenous people living in the forest margins (Uniyal et al., 2005).

Despite the diverse nature of crops grown in the country and the existence of a fast

growing pharmaceutical sector, the share of India in world trade is quite insignificant considering

the large geographical area. However, this is bound to rise rapidly with better research inputs and

efficient management of the farm sector. So far, India has been involved in the export of only

large volume raw material. To achieve competitive advantage we need to resort to low volume

high cost (value) trade through value addition to the raw and unfinished products. It is therefore,

necessary to develop genetically superior planting material for assured uniformity and desired

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quality and resort to organized cultivation to ensure the supply of raw material at grower‟s end.

Post harvest storage and process technologies need to be developed to produce the value added

finished products that may be directly utilized by the industry.

1.5 Portulaca quadrifida Linn.

Portulaca quadrifida Linn. belongs to the family portulacaceae. It is a small diffused,

succulent, annual herb found throughout the tropical parts of India. The leaves of Portulaca

quadrifida Linn. are elliptic, oval or ovate lanceolate, 3-6 mm long, flowers yellow, solitary

terminal, petals 4 yellow, stamens 8-10, capsules conical seeds minutely tubercled. Herb annual

stem articulated, prostrate, rooting at nodes, leaf axils patent pilose. Petiole is absent, ovary

ovoid, stigma 3 or 4 lobed, seeds gray, subglobose, laterally compressed, minute municate

(Plate- 1)

The plant is commonly called as Chicken weed, Wild purslane or Ten „o‟ clock plant in

English, kulfa in Hindi, Goni soppu in Kannada, Pasalai keerai in Tamil, Goddu pavelli in

Telugu.

P. quadrifida Linn. is found in wild on bare patches and among rocks, on sandy or stony

soils, from sea-level up to 2000 m altitude. It is found throughout the tropical and subtropical

parts of India and Africa. It is often involuntarily introduced by the agency of man and readily

occupies newly disturbed areas, compost and rubbish heaps and fields. In some African

languages it is called „Lord of the rubbish heap‟. It is tolerant of a wide range of soils but

prefers sand or sandy loams. The wide distribution and large variation of P. quadrifida Linn.

points to great genetic flexibility that rapidly permits adaptation to new environments.

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Plate 1: Portulaca quadrifida Linn. plant

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 P. quadrifida Linn. is used as a vegetable and also used for various curative purposes. It

is said to be useful in asthma, cough, urinary discharges, inflammations and ulcers. A poultice of

the plant is applied in abdominal complaints, erysipelas and haemorrhoids (Kirtikar and Basu,

2001). The leaves and young shoots of P.quadrifida Linn. are collected from the wild and are

eaten raw. They have a mild pleasant flavour and are frequently used in salads. They are also

consumed as a cooked vegetable. In India boiled leaves are mixed with sorghum or pearl millet

flour in preparing a kind of bread. Plants are a good feed for chicken and other birds. Large types

are sometimes planted as an ornamental (e.g. in Rwanda) or as a soil binder to prevent erosion

(e.g. in Kenya). Medicinally P. quadrifida Linn. is used less widely but has similar medicinal

applications as Portulaca oleracea L. The general uses are as a diuretic, to treat rheumatism and

gynaecological diseases, as a sedative, analgesic and cardiotonic, to treat fever, disorders of the

urinary tract, worm diseases, as a tonic and choleretic, to treat dysentery, and to apply externally

to ulcers, eczema and dermatitis (Burkill, 1997). P. quadrifida Linn. has been reported to

possess antifungal activity against Aspergillus fumigates and Candida albicans ( Hoffman et al.,

2004).

Portulaca comprises about 150 species, of which about 30 occur in tropical Africa, but

opinions on species delimitation differ considerably. Particularly the group of species with

alternate leaves is taxonomically difficult. P. quadrifida Linn. is rather unique with its mat-

forming habit, but it is very variable, particularly in stamen number and flower size. In Africa

only Portulaca pilosa L. has a similar habit, but it has alternate leaves, 5 petals and more

numerous stamens. Like Amaranthus, Portulaca is characterized by the C4-cycle photosynthetic

pathway, which means a high rate of photosynthesis at high light intensity and temperatures.

Fresh seeds need light for germination, but this requirement disappears in older seeds.

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Generative development seems not to be influenced by photoperiod. The flowers are said to open

promptly at 10 a.m., hence the English name „ten o‟clock plant‟. Seeds of P. quadrifida Linn. are

easily spread by wind, water, with crop seeds or through bird droppings, and as a weed it is

difficult to control because it also easily propagates from small fragments (Gilbert and Phillips,

2000).

1.6 Some related species of Portulaca

Portulaca oleracia

Known as Purslane or Ma Chi Xian (pinyin: translates as "horse tooth amaranth") in

traditional Chinese medicine, its active constituents include: noradrenaline, calcium salts,
dopamine, DOPA, malic acid, citric acid, glutamic acid, asparagic acid, nicotinic acid, alanine,
glucose, fructose, and sucrose (Tierra and Michael, 1988). Betacyanins isolated from P.oleracea
improved cognition deficits in aged mice. A rare subclass of homoisoflavonoids, from the plant,
showed in vitro cytotoxic activities towards four human cancer cell lines. Use is contra indicated
during pregnancy and for those with cold and weak digestion. Purslane is a clinically effective
treatment for oral lichen planus, and its leaves are used to treat insect or snake bites on the skin,
boils, sores, pain from bee stings, bacillary dysentery, diarrhea, hemorrhoids, postpartum
bleeding, and intestinal bleeding (Wang and Yang , 2010).

P. oleracea efficiently removes bisphenol A, an endocrine-disrupting chemical, from a

hydroponic solution. How this happens is unclear (Watanabe et al., 2012).

Portulaca glandiflora
Portulaca grandiflora Hook. (Portulacaceae) is a succulent plant. The plant is used for

the cure of sore throat and skin rashes. It is a putative immunostimulant (Zheng and Zhang,

1990). It is also used for detoxification. P. grandiflora has been reported for its efficacy on

hepatitis B surface antigen. In addition, antimutagenic effect on the mutation induced by

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alfatoxin B1 and cyclophosphamide in mice has been demonstrated. Aerial parts of P.

glandiflora are reported to contain various diterpenoids like portulal, portulenone, portulenol,

portulene and portulene acetal a minor diterpenoid ( Ohsaki et al., 1997).

1.7 Aim and objectives of the present work

It is evident from the literature, in Indian sub-continent, the whole plant material of

P.quadrifida Linn. is used in the traditional medicine to cure innumerable ailments, without any

scientific knowledge. In spite of its various medicinal uses very little work has been carried out

on this plant. Present investigation was undertaken to investigate detailed phytochemical and

pharmacological aspects of P. quadrifida Linn. with following objectives.

 Preliminary phytochemical screening of various secondary metabolites from the

different extracts of P. quadrifida Linn.

 Standardisation of the plant material by pharmacognostic studies.

 Separation of bioactives and their identification by HPLC.

 Evaluation of antioxidant properties of the plant.

 Evaluation of anticancer properties of the plant.

 Evaluation of neuropharmacological properties of the plant.

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