1

INTRODUCTION

“There is no plant in the Universe which is non medicinal and which can be

made use of for many purposes and by many modes” (Anonymous, 1951). This

definition rightly suggests that in principle all plants have a potential medicinal value.

Medicinal plants have been considered as important therapeutic aid for alleviating

ailments of humankind. Search for eternal health and longevity and to seek remedy to

relieve pain and discomfort promoted the early man to explore his immediate natural

surroundings to develop a variety of therapeutic agents using natural resources. Herbs

are staging a comeback and herbal ‘renaissance’ is happening all over the globe. The

herbal products today symbolize safety in contrast to the synthetics that are regarded

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

medicinal, flavoring 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 back to the naturals with a hope of

safety and security.

Hundreds if not thousands of indigenous plants have been used by man from

prehistoric times on all continents for relieving suffering and curing ailments. The

practice of organized herbal medicine dates back to the earliest periods of known

human history. Medicinal plants have been used in the treatment of diseases in almost

all ancient civilizations, from 3700 B.C. whether it is Egypt or Chinese, the Greeks or

the Romans. The Petric collection from Kahun in Egypt (1880 B.C.), Atharvaveda

(1200 B.C) from India and the Avesta (6 A.D) from Persia show that the early

medicine was based mainly on religion and magic but also included a growing use of

herbs.

Inspite of tremendous development in the field of allopathy, medicinal plants

and their derivatives still remain one of the major sources of drugs in modern and

traditional systems throughout the world playing a major role in medicinal therapy.

Medicinal plants form the back bone of traditional medicine and hence more than

3300 million people utilize medicinal plants on a regular basis. Demand for medicinal

plants is increasing due to growing recognition of natural products being non toxic,

having no side effects. Furthermore an increasing reliance on the use of medicinal

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plants in the industrialized societies has been traced to the extraction and development

of several drugs and chemotherapeutics from the plants as well as from traditionally

used rural remedies. Moreover in these societies herbal remedies have become more

popular in the treatment of minor ailments on account of the increasing cost of

personal health maintenance. The WHO estimates that 80% of people living in

developing countries rely almost exclusively on traditional medicine for their primary

health care needs. 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 Asian countries such as China and India, the contribution is as much as

80%.

India is well known as an Emporium and a rich repository of medicinal plants.

Knowledge of medicinal use of plants in India is amassed over millennia by tribes.

For thousands of years Indian plants have been attracting attention of foreign

countries. People from countries like China, Cambodia, Indonesia and Baghdad used

to come to ancient Universities of India like Takshila (700 B.C) and Nalanda (500

B.C) to learn health science of India. India is a treasure chest of biodiversity which

host large variety of plants and has been identified as one of the eight important

Vavilorian centers of origin and crop diversity. Wide variation in climatic,

meteorological and topographical conditions prevailing in India due to its vastness

makes it the repository of perhaps the most varied and luxuriant flora growing

anywhere on the surface of the earth. Indian flora is not only rich but very

cosmopolitan with the presence of over 45000 different plant species (Joy et al.,

1998). India’s diversity is unmatched due to the presence of 16 different agro-climatic

zones, 10 vegetation zones, 25 biotic provinces and 426 biomes. Of these, about

40000 plants have good medicinal value. However, only 7000-7500 species are used

for their medicinal value by traditional communities. In India, drugs of herbal origin

have been used in traditional systems of medicine such as Unani and Ayurveda since

ancient times and they still serve as classical formulations, in the present system of

medicine. The Ayurveda system of medicine uses about 700 species, Rigveda (500

BC) has recorded 67 medicinal plants, Yajurveda (1400-1000 BC) with 81 species,

Atharvaveda(4500-2500 BC)with 290 species, Charaka Samhita (700 BC) has

described about 1100 species, Sushruta Samhita (200 BC) has description of about

1270 species, Unani 700, Siddha 600, Amchi 600 and modern medicine uses about

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700 species (Joy et al., 1998). Unfortunately, much of the ancient knowledge and

many valuable plants are being lost at an alarming rate. With the rapid depletion of

forests, impairing the availability of raw drugs, Ayurveda, like other systems of herbal

medicines has reached a very critical phase. About 50% of the tropical forests, the

treasure house of plant and animal diversity have already been destroyed. In India,

forest cover is disappearing at an annual rate 1.5 mha/yr. What is left at present is

only 8% as against a mandatory 33% of the geographical area (Joy et al., 1998). Many

valuable medicinal plants are under the verge of extinction.

As Botanist Walter Lewis and Microbiologist Memory Elvin Lewis (1982),

put in their book Medical Botany: “Nature is still mankind’s greatest chemist and

many compounds that remain undiscovered in plants are beyond the

imagination of even our best scientists”. Apart from being the sources for new drug

the plants continue to play an important role in modern therapy.

Inspite of rapid development in the methods of organic synthesis in

laboratories, medicinal plants continue to play a significant role in modern medicine

and serve as model in drug development due to their inherent distinct chemical and

biological properties. In nature a plant is able to synthesize complex molecules,

namely alkaloids, terpenoids, tannins, saponins, glycosides, etc., collectively called

secondary metabolites, from simple ones through highly specific reaction mechanisms

that they use for defense and communication. It is difficult and expensive to duplicate

such synthesis in laboratory. The compounds synthesized by the plants play an

important role as medicinal and pharmaceutical agents not only as purified isolates

and extractives but also as lead compounds for synthetic optimization. Plants, the

small fraction of flowering plants that have so far been investigated have yielded

about 120 therapeutic agents of known structure from about 90 species of plants. The

drugs are derived either from the whole plant or from different organs, like leaves,

stems, bark, root, flower, seed, etc (Cragg and David, 2001). Some drugs are prepared

from excretory plant products such as gum, resins and latex. Even Allopathic system

of medicine has adopted a number of plant derived drugs which form an important

segment of the modern pharmacopoeia.

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Some of the useful plant drugs include vinblastine, vincristine, taxol,

podophyllotoxin, camptothecin, digitoxigenin, gitoxigenin, digoxigenin, tubocurarine,

morphine, codeine, aspirin, atropine, pilocarpine, capscicine, allicin, curcumin,

artemesinin and ephedrine among others. In some cases, the crude extract of

medicinal plants may be used as medicaments. On the other hand, the isolation and

identification of the active principles and elucidation of the mechanism of action of a

drug is of paramount importance. Hence work in both mixture of traditional medicine

and single active compounds are very important, where the active molecule cannot be

synthesized economically, the product must be obtained from the cultivation of plant

material. As the plant derived drugs not only offers a stable market worldwide, but

also plants continue to be an important source for new drug. About 121 (45 tropical

and 76 subtropical) major plant drugs have been identified for which no synthetic one

is currently available. Some of the plant drugs for which no synthetic one is currently

available is listed in Table-1.The scientific study of traditional medicines, derivation

of drugs through bioprospecting and systematic conservation of the concerned

medicinal plants are thus of great importance.

Many programmes have been initiated for in situ propagation of medicinal

plants in recent times by Non government organisations as well as government and

semi-government agencies. However, these programmes, though laudable are limited

by several factors. The potential for regeneration of many plants in their natural

habitat is poor. The germination of seeds and establishment of seedlings is also poor.

There is a very little knowledge of the vegetative propagation of these plants.

Hence tissue culture technology is utilized for the conservation and mass

propagation of selected native perennial plants that cannot be propagated on a large

scale by means of seeds and cuttings.

During the past two decades, there has been a great interest and progress in in

vitro propagation of medicinal plants using techniques of tissue, meristem, protoplast

and organ culture.

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Table: 1. The major plant drugs for which no synthetic one is currently available (Kumar et al., 1997).

Drug Plant Use

Allicin Allium sativum Antifungal, amoebiasis

Amalacine Catharanthus roseus Anticancer, hypotensive

Artemisinin Artemesia annua Antimalarial

Atropine Atropa belladona Spasmolytic, cold

Atropine Hyoscyamus niger Spasmolytic, cold

Berberine Berberis For leishmaniasis

Cardiac glycosides Digitalis sp. For congestive heart failure

Catechin Acacia catechu Antiulcer

Cocaine Erythroxylum coca Topical anaesthetic

Codeine Papaver somniferum Anticough

Diospyrin Diospyros Montana

Emetine Cephaelis ipecacuanha Amoebiasis

Forskolin Coleus forskohlii Hypotensive, cardiotonic

Glycyrrhizin Glycyrrhizia glabra Antiulcer

Gossypol Gossypium sp. Antispermatogenic

Magnolol Magnolia bank Peptic ulcer

Nimbidin Azardichta indica Antiulcer

Pilocarpine Pilocarpus jaborandi Antiglaucoma

Plumbagin Plumbago indica Antibacterial, antifungal

Pristimerin Celastrus paniculata Antimalarial

Quassinoids Ailanthus Antiprotozoal

Quinine Cinchona sp. Antimalarial, amoebic dysentery

Rescinnamine Rauwolfia serpentina Tranquilizer

Reserpine Rauwolfia serpentina Tranquilizer

Ricin Ricinus communis

Sophoradin Sophora subprostrata Antiulcer

Taxol Taxus baccata

Taxus brevifalia

Breast and ovary cancer, antitumour

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Vinblastine Catharanthus roseus Anticancer

Need for In vitro propagation

The plant cell, tissue and organ culture opened up new possibilities in improvement

and micropropagation of desired elite strains of fruits, ornamentals, vegetatively

propagated agri-horticultural, medicinal and plantation crops. In vitro regeneration is

an efficient means of ex-situ conservation of medicinal plants (Fay, 1994). The

primary goal of in vitro culture of medicinal plants has always been mass clonal

propagation or micropropagation of the most desirable genotypes.

Micropropagation is the most significant and conventional method which is

widely used in commercial production of plants (Morel, 1960). It gives many benefits

to the breeders such as, increase in propagation rate of plants, availability of plants

throughout the year and conservation of genetic resources (Bajaj et al., 1988). As a

result, plant tissue culture has acquired many practical applications in agriculture and

industry. Most of the dicotyledonous and monocotyledonous plants have been

cultured or micropropagated by in vitro technique and over 1000 species have been

conserved using this technique (Brown and Thorpe, 1986; George and Debergh, 2008

and Kane et al., 2008). The process of rejuvenation (return to juvenile state)

successfully carried out by organogenesis and somatic embryogenesis.

Organogenesis:

Organogenesis is a process of differentiation by which plant organs like shoots, roots,

flower buds etc., are formed. The process of organogenesis is either through callus

formation (indirect organogenesis) or directly from explants (direct organogenesis).

Differentiation of plants from callus cultures has been suggested as a potential method

for rapid propagation and for induction of variations (Martin, 2002).

Somatic embryogenesis:

Somatic embryogenesis is the development of embryo from somatic cells, tissues or

organs. They are analogous to their zygotic counter-parts. The process differs from

organogenesis where the embryo being the bipolar structure with a closed radicular

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end and plumule end unlike that of the monopolar structure. Embryo differentiation is

considerably influenced by the physiological state of the calli themselves and the

carry over effects of auxins from inoculums to subculture. The embryoids pass

through the same sequential stages of embryos during the development into plantlet

(Tejavathi et al., 2000 and Chawla et al., 2002).

In vitro mutagenesis:

Mutations, the heritable changes in the genetic material are the ultimate source of all

genetic variations between individuals. Mutation breeding is an important tool in crop

improvement of vegetatively propagated crops and particularly in plants with

reproductive sterility (Broertjes and Van Harten, 1988). The practical use of this

method is reflected in the number of mutant cultivars evolved and put into cultivation.

Advent of in vitro techniques has opened new avenues in improvement of crop plants.

The development of efficient in vitro culture methods has facilitated the use of

mutation technique for improvement of both seed and vegetatively propagated plants.

In many vegetatively propagated crops, induction of mutation in combination with in

vitro culture techniques is the only effective method for crop improvement (Novak et

al., 1990). Induced mutations are being used for analyzing the effect of various types

of known alteration in DNA on expression of the concerned characters.

Induced mutation technique is a valuable tool and usually exploited in plant

breeding. Tissue culture makes it more efficient by allowing the handling of large

populations and by increasing mutation induction efficiency, possibility of mutant

recovery and speediness of cloning selected variants. Some vegetatively-propagated

species are recalcitrant to plant regeneration, which can be a limit for the application

of gene transfer biotechnology, but not for mutation induction breeding. Mutagenesis

offers the possibility of altering only one or a few characters of an already first-rate

cultivar, while preserving the overall characteristics (Sharma et al., 2005). Traits

induced by mutagenesis include plant size, blooming time and fruit ripening, fruit

colour, self-compatibility, self-thinning, and resistance to pathogens (Predieri, 2001).

The combination of in vitro culture and mutagenesis is relatively inexpensive, simple

and efficient (Ahloowalia, 1998). The availability of suitable selection methods could

improve its effectiveness and potential applications. The molecular marker technology

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available today already provides tools to assist in mutation induction protocols by

investigating both genetic variation within populations and early detection of mutants

with desired traits. However, cost still represents a major limitation to their

application.

For any mutation breeding programme, selection of effective and efficient

mutagen is very essential to recover higher frequency of desirable mutations (Kawai,

1975, 1986; Shah et al., 2008 and Girija and Dhanavel, 2009). Effectiveness usually

means the rate of point mutations relative to dose, whereas efficiency refers to the rate

of point mutations relative to other biological effects induced by the mutagen and is

considered a measure of damage (Konzak et al., 1965).

A number of chemical and physical mutagens are widely employed to induce

genetic variability in plants. Among the techniques and sources of genetic variation

available for tissue culture mutation induction, physical mutagens have already shown

potential for application in plant breeding. The types of radiation suitable for

mutagenesis are ultraviolet radiation (UV) and ionizing radiation (X-rays, gamma-

rays, alpha and beta particles, protons, and neutrons) (Brunner, 1995; Bhatia et al.,

2001; Irfaq and Nawab, 2003; Joseph et al., 2004; Sangsiri et al., 2005 and Tah,

2006). X-rays and gamma-rays are the most convenient and easiest types of radiation

to use with regards to application methods and handling (Sanada and Amano, 1998),

and have been both the most widely used ionizing radiation types and the most

effective for plant breeding purposes. At present, Gamma rays are the most favoured

physical mutagenic agent (Solanki, 2005) and it is mainly used to prolong the

preservation time (half time) of food to prevent reproduction or to kill pathogenic

microorganisms. Gamma ray doses below a certain threshold value (1.6 R/h) are

considered fully safe to human health as no radioactivity remains in the irradiated

materials. It is usually obtained by the disintegration of radioisotopes as 60Cobalt, it is

usually electron emitter and can be brought into cell culture or specific plant section

in order to study physiological process. A number of chemicals have been found to be

equally and even many times more effective and efficient mutagens (Thakur and

Sethi, 1995; Kharkwal, 1998; Rekha and Langer, 2007; Basu et al., 2008, Dhanavel et

al., 2008; Ganapathy et al., 2008 and Wani, 2009).

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Furthermore, physical mutagens have some technical advantages over

chemical mutagens. With regards to safety and environmental issues there is no need

for manipulation of hazardous substances and production of toxic residues. Physical

mutagen post-treatment manipulation is simpler and allows for a more precise

determination of exposure time. Ethyl Methane Sulphonate (EMS) is one of the

favoured chemical mutagens used in mutational studies (Bhatia, 1967). EMS is a

colourless liquid compound with molecular weight of 124 and is very reactive

compound. As consequence, solution should be prepared just before use depending on

the required concentration. EMS is an efficient chemical mutagen in higher plants. It

acts directly by virtue of its ability either to modify a particular base of DNA or to

become possible to induce many changes in any gene. As a result, susceptibility to

mutation is roughly proportional to the size of the genes (Benjamin, 2000) and

exclusively produces GC to AT transition, which alters the ability of bases to form

normal base pairs in eukaryotic organism. Hence physical mutagenic agent like

gamma radiation and chemical mutagenic agent like EMS proved to be useful in

extending the existing variability under in vitro condition.

Phytochemical analysis

Herbal medicines have become more popular in the treatment of many diseases due to

popular belief that green medicine is safe, easily available and with fewer side effects.

Various medicinal properties have been attributed to natural herbs. Medicinal plants

constitute the main source of new pharmaceuticals and healthcare products (Ivanova

et al, 2005). The use of medicinal plants in the industrialised societies has been traced

to the extraction and development of several drugs from these plants as well as from

traditionally used folk medicine (Shrikumar and Ravi, 2007). Extraction and

characterization of several active phytocompounds from these green factories have

given birth to some high activity profile drugs (Mandal et al, 2007). The use of

traditional medicine is widespread in India (Jeyachandran and Mahesh, 2007). A

growing body of evidence indicates that secondary plant metabolites play critical

roles in human health and may be nutritionally important (Hertog et al., 1993). It is

believed that crude extract from medicinal plants are more biologically active than

isolated compounds due to their synergistic effects (Jana and Shekhawat, 2010),

Phytochemical screening of plants has revealed the presence of numerous chemicals

including alkaloids, tannins, flavonoids, steroids, glycosides, vitamins, amino acids

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and saponins etc. Any part of the plant may contain active components. Knowledge of

the chemical constituents of plants is desirable because such information will be of

value for the synthesis of complex chemical substances. Such phytochemical

screening of various plants is reported by many workers (Mojab et al., 2003; Parekh

and Chanda, 2007 and Parekh and Chanda, 2008).

The phenolic compounds are one of the largest and most ubiquitous groups of

plant metabolites that possess an aromatic ring bearing one or more hydroxyl

constituents (Singh et al., 2007). Phenolic compounds are widely found in the

secondary products of medicinal plants, as well as in many edible plants (Hagerman et

al., 1998). A number of studies have focused on the biological activities of phenolic

compounds, which are potential antioxidants and free radical-scavengers (Rice-Evans

et al., 1995; Cespedes et al., 2008; Reddy et al., 2008 and Chanda and Dave, 2009).

They posses biological properties such as: anti-apoptosis, anti-ageing, anti-

carcinogen, anti-inflammation, anti-atherosclerosis, cardiovascular protection and

improvement of the endothelial function, as well as inhibition of angiogenesis and cell

proliferation activity (Han et al., 2007). Several studies have described the antioxidant

properties of medicinal plants, foods and beverages which are rich in phenolic

compounds (Brown and Rice-Evans, 1998 and Krings and Berger, 2001). Natural

antioxidants mainly come from plants in the form of phenolic compounds such as

flavonoids, phenolic acids, tocopherols, etc. (Ali et al., 2008). Many reports suggest

that plants which are having more phenolic content show good antioxidant activity

that is there is a direct correlation between total phenol content and antioxidant

activity (Brighente et al., 2007 and Salazar et al., 2008).

Normally free radicals of different forms are generated at a low level in cells

to help in the modulation of several physiological functions and are quenched by an

integrated antioxidant system in the body. However, if free radicals are produced in

excess amount they can be destructive leading to inflammation, ischemia, lung

damage and other degenerative diseases (Halliwell et al., 1992 and Cavalcanti et al.,

2006), Free radical reactions, especially with participation of oxidative radicals, have

been shown to be involved in many biological processes that cause damage to lipids,

proteins, membranes and nucleic acids, thus giving rise to a variety of diseases (Lee et

al., 2005 and Campos et al., 2006).

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Secondary metabolic compounds are characterized by High Performance

Liquid Chromatography (HPLC). It is a chromatographic technique that can separate

a mixture of compounds and is used in biochemistry and analytical chemistry to

identify, quantify and purify the individual components of the mixture. It is based on

the principles of adsorption, partition, ion-exchange, exclusion and affinity

chromatography. HPLC technique is used with high pressure (up to 8,000 psi). The

flow rate is high and experimental time is shortened considerably. Therefore, this

technique is highly efficient and has a very fast speed of resolution.

Inspite of rapid development in use of medicinal plants there is still lacunae

when it comes to proper use and knowledge of negativity associated with them. Hence

modern approaches of science and technology should be used to overcome such

negativities or drawbacks for tremendous progress in the field of herbal medicines.

In the true sense the actual ‘Revolution’ in the field of herbal medicine takes

place when any plant not only serves for medicinal value but also satisfies the basic

needs of humankind like food and nutrition because in the present world “FOOD,

NUTRITION AND MEDICINE” has become the most essential commodities for a

life to sustain. Great people have farsighted the importance of food and medicine –

“Let the Food be the medicine and the medicine be the food.”

-Hippocrates, 400BC

“The doctor of the future will give no medicine, but will interest his patient in the

care of the human frame, in Diet and the cause and prevention of disease.”

-Thomas Edison.

One such plant which aptly goes with these words is Sauropus androgynous.

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About the Plant

Sauropus androgynous (L.) Merr.

Systematic position

Kingdom Plantae

Division Magnoliophyta

Class Magnoliopsida

Order Malpighiales/Euphorbiales.

Family Phyllanthaceae/Euphorbiaceae

Genus Sauropus

Species androgynous.

Distribution:

Distributed in South Asia and Southeast Asian countries including China, India,

Srilanka, Indo-China, Indonesia, Malaysia, Philippines, etc.

Vernacular Names:

Malaysia Asin-asin, Cekor- Manis, Cekup- Manis, Cermela- hutan, Taruk -

Manis, Katuk.

English Star Goose Berry.

Indonesia Babing, Gerager, Kerakur, Memata, Simani.

Thailand Phak -Waan.

Philippines Binahian, Malunggay- hapon.

Vietnam Rau- nyout, Bongot.

Laos Hvaan –baanz.

China Ma-Ni-Chai, So-Kun-Mu, Fan-Shu-Choy, Pa-Wan.

Japan Ruridama-no ki.

India Kutchumku, Surasarabi (Hindi); Aruni (Sanskrit); Chakramuni

(Kannada); Thavasai murungai (Tamil).

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Propagation:

Propagation is by stem cuttings and seeds. The plants are mainly propagated

vegetatively in commercial planting approximately 20 cm long, woody stem cuttings.

The cuttings are placed spacing 30 cm long and 30 cm wide. After reaching 50-60 cm

high, the plants are pruned in order to get the young and fresh leaves. Seed

propagation is quite rare due to poor fruit set and short viable period of seeds.

Parts used:

Whole plant including, tender leaves, shoot tips, flowers, immature fruits and roots.

Predominantly leaves are consumed as green vegetables. They are eaten boiled, stir

fried or in soups. They are added to sandwiches, meat, rice, curries, scrambled eggs,

etc., used also to garnish.

Nutritional value:

Sauropus androgynous is highly nutritious; protein content is higher than other leafy

vegetables. Fresh leaves are excellent source of antioxidants (Hemalatha et al., 1999;

Asmah Rahmat et al., 2003 and Mahuya De Ghosh et al., 2011), carotenoids (Tee et

al., 1992), pro vitamin A, Vitamin B, C, D, E (Ling Soon Ching et al., 2001). It is

among only a few flora containing vitamin K. Leaves are also rich in carbohydrates

and minerals like calcium, potassium, phosphorus, copper, iron, sodium and are rich

sources of fibre (Padmavathi et al., 1990 and Singh et al., 2011).

Apart from these the leaves also contain essential amino acids like: Lysine,

methionine, tryptophan, phenylalanine, threonine, valine, leucine, iso leucine and

nicotinic acid (Prajapati et al., 2010).

Other chemical constituents include:

Lutein and Zeaxanthin (Liu-Ye Ting et al., 2007).

Ligan glycosides like – Ligan di glycoside, (-)-isolariciresinol 3 alpha-O-beta-

apiofuranosyl-(1->2)-O-beta-glucopyranoside (Kanchanapoom, 2003).

Megastigmane glycosides, sauroposide, (+)-isolariciresinol 3 alpha-o-beta-

glucopyranoside, (-)-isolariciresinol 3 alpha-O-beta-glucopyranoside, (+) -

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syringaresinol di-O-beta-glucopyanoside, guanosine and corchoionoside C were

extracted from aerial parts of Sauropus androgynous (Kanchanapoom, 2003).

Uses:

Culinary uses:

Predominantly used as “Green leafy nutritious vegetable “due to high yield and

palatability. The shoot tips have been sold as “tropical asparagus”. The leaves and

the top 15cm of the stem tips of the Sauropus plant have a pleasant taste, similar to

fresh garden peas, with slightly nutty flavour. They are normally eaten raw in salads

or steamed, added to stir-fry, rice and egg dishes, soups or casseroles. In Vietnam, the

locals cook it with crab meat, minced pork or dried shrimps to make soup. In

Malaysia, it is commonly stir-fried with eggs or dried anchovies. The leaves retain

their dark green colour and firm texture on cooling.

Fruits are candied.

A green dye obtained from leaves is used as food colouring for pastries, rice and

preserves.

Traditional uses for the treatment of diseases:

Genito-urinary diseases: The roots of Sauropus androgynous, have diuretic properties

which is being taken advantage of by some traditional practitioners to treat various

urinary complaints. Amongst the diseases treated include, symptomatic relieve of

dysuria (Hean Chooi Ong, 2003).

Cardiovascular diseases: Again the roots are used in the treatment of cardiovascular

disease or symptoms of this group of diseases including vertigo, dizziness, fainting

spells. It is also being recommended to those with hypertension (Hean chooi Ong,

2003).

Antiobesity: The leaf extract of Sauropus androgynous was used for body weight

reduction. Its popularity as a “Slimming agent” was high in Asian countries

specifically in Taiwan, Malaysia, etc., in mid 90’s. In America, since 1995, Sauropus

androgynous leaves fries, salads and beverages were consumed by many people as

drug antiobesitas (body slimming).

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Other uses:

The leaves are given to women after delivery to allow post partum recovery

and to enhance lactation in feeding mothers. The enhancement in the breast

milk production probably derived from the hormonal effects of chemical

compounds that are estrogenic sterols (Sa’ roni et al., 2004)

The leaf juice is used to treat cholecystosis, diorrhea and other forms of fever,

rhinosis (Timothy Johnson, 1998).

Decoction of the leaves and roots is remedy for epitaxis and oriental sores

(John Harry Wiersema, 1999)

Another application of the leaves is for oral thrush in infants (Koh Hwee Ling

et al., 2009). Paste of the leaves is applied over nasal ulcers and yaws

(Timothy Johnson, 1998), erythrema and measles.

Juice extracted from the leaves is used as an eye lotion for eye complaints.

(Kanchanapoom et al., 2003).

Leaves are used as cattle and poultry feed.

Planted as a live fence in home gardens.

Drugs:

Acivit- Is a capsule given as supplement for breast feeding. Each capsule of ACIVIT

is said to contain Sauropus androgynous extract along with Cucurma xanthorriza

extract,Spirulina, multivitamin and minerals. It is said to be an herb medicine that

helps to obtain and maintain optimum health.

Toxicity associated with S.androgynous:

Bronchiolitis Obliterans:

The excessive intake of Sauropus androgynous for weight reduction led to an

outbreak of lung damage manifested by a condition called “Bronchiolitis

Obliterans” (Lai et al., 1996; Lin et al., 1996; Chang et al., 1997 and Ger et al.,

1997). The damage was so severe that there were even cases of lung transplantation

(Luh et al., 1999). This was said be associated with an alkaloid called “Papaverine”

which is an isoquinoline alkaloid used as vasodilator during cardiac surgeries to

smoothen the heart muscles.

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Leaves of Sauropus androgynous contain considerable amount of the alkaloid

papaverine (580 mg/100gm fresh leaves). Study report established between the

association and consumption reveals that the people consumed large amount (Average

per week, 814±417g) i.e., much higher than the restricted dose (Hsiue et al., 1998 and

Wu et al., 1998). Reports also say that people consumed uncooked juice, rather than

the traditional boiled and stir fried form of preparation, this led to the possible

disproportion of matrix metalloproteinase/ tissue inhibitor of metalloproteinase

occurred in the bronchiole local field and might be involved in the disease state

formation of Bronchiolitis Obliterans.

Further study results indicate that necrosis and apoptosis are involved in the

toxic effect of S.androgynous in NIH3T3 fibroblasts (Chang et al., 1998). However,

more evidence is needed to clarify if necrosis and apoptosis are also related to the

pathogenesis of S. androgynous associated Bronchiolitis Obliterans.

Anaphylactic reaction: Reports (Stirapongsasuti et al., 2010) for the first time

reported anaphylactic reaction in one patient with latex allergy, following the

ingestion of S.androgynous.

Other adverse effects in Human: Consuming too much of the leaves can cause pains

in the limbs, breathing difficulties, dizziness, diplopia, nausea and weakness

(Padmavathi et al., 1990).

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Main active principle:

PAPAVERINE:

Name: Papaverine,

IUPAC name: 1-(3.4-dimethoxybenzyl)-6,7-dimethoxyisoquinoline,

Trade name: Pavabid,

Molecular formula: C20H21NO4,

Molecular Structure:

CAS number: 58-74-2, 61-25-6(hydrochloride),

Molar mass: 339.385 g/mol.

Is a Benzyl-isoquinoline alkaloid, basically an opium alkaloid used as antispasmodic

drug.

Uses:

Papaverine is primarily used in the treatment of visceral spasm, vasospasm (especially

those involving the heart and the brain). While it is found in opium poppy, papaverine

differs in both structure and pharmacological action from the analgesic (morphine-

related) opium alkaloids (opioids).

It is approved to treat spasms of the gastrointestinal tract, bile ducts and ureter

and for use as a cerebral and coronary vasodilator (Bella et al., 2004) in subarachnoid

haemorrhage combined with balloon angioplasty (Muller Schweinitzer et al., 1993)

and coronary artery bypass surgery (Brockbank, 1994). Papaverine may also be used

as a smooth muscle relaxant in microsurgery where it is applied directly to blood

vessels. Also used in the treatment of erectile dysfunction, either alone or in

combination with other drugs (Desvaux, 2005).

18

It is commonly used in cryopreservation of blood vessels along with the other

glycos-aminoglycans and protein suspensions (Muller Schweinitzer et al., 1992

and1993). Functions as a vasodilator during cryopreservation when used in

conjunction with verapamil, phentolamine, nifedipine, tolazoline or nitroprusside

(Brockbank, 1994 and Giglia et al., 2002).

Papaverine is also being investigated as a topical growth factor in tissue

expansion with some success (Tang et al., 2004).

It is used as an off label prophylaxis (preventative) of migraine headaches

(Sillanpaa et al., 1978; Vijayan, 1977 and Poser, 1974).

Mechanism and Side effects:

The in vivo mechanism of action is not entirely clear, but an inhibition of the enzyme

phospho-di-esterase causing elevation of cyclic AMP levels is significant. It may also

alter mitochondrial respiration. It has also been demonstrated to be a selective

phospho-di-esterase inhibitor for the PDE10A subtype found mainly in the striatum of

the brain. When administered chronically to mice, it produced motor and cognitive

deficits and increased anxiety, but conversely may produce an antipsychotic effect

(Siuciak et al., 2006 and Hebb et al., 2008), even though not all studies support this

view (Weber et al., 2009).

Frequent side effects of papaverine treatment include polymorphic ventricular

tachycardia, constipation, interference with sulpho-bromophthalein (Giglia et al.,

2002) retention test (used to determine hepatic function), increased transaminase

levels, increased alkaline phosphatise levels, somnolence and vertigo (Bella et al.,

2004).

Rare side effects include flushing of the face, hyperhidrosis (excessive

sweating), cutaneous eruption, arterial hypotension, tachycardia, loss of appetite,

jaundice, eosinophilia, thrombopenia, mixed hepatitis, headache, allergic reaction,

chronic active hepatitis (Bella et al., 2004) and paradoxical aggravation of cerebral

vasospasm (Tang et al., 2004).

19

Formulations and trade names:

Papaverine is available as a conjugate of hydrochloride, codecarboxylate, adenylate

and teprosylate (Sillanpaa et al., 1978). It was also once available as a salt of

hydrobromide, camsylate, cromesilate, nicotinate and phenylglycolate. The

hydrochloride salt is available for intramuscular, intravenous, rectal and oral

administration (Muller Schweinitzer et al.1993). The teprosylate is available in

intravenous, intramuscular and orally administered formulations (Vijayan, 1977). The

codecarboxylate is available in oral form only (Poser, 1974) as is the adenylate

(Siuciak et al., 2006).

The code-carboxylate is sold under the name Albatran (Hebb et al., 2008), the

adenylate as Dicertan (Weber et al., 2009), and hydrochloride salt is sold variously as

Artegodan (Germany), Cardioverina and Dispamil (countries outside Europe and the

United States), Opdensit (Germany), Panergon (Germany), Paverina Houde (Italy,

Belgium), Pavacap and Pavadyl (United States), Papaverine (India and Israel)

Papaverin-Hamelin and Spasmo-Nit(Germany) (Muller Schweinitzer et al., 1992),

Cardiospan, Papaversan, Cepaverin, Cerespan, Drapavel, Forpaven, Papalease,

Pavatest, Paverolan, Therapav (Quebec).

Nutrition Value:

Nutrition also called nourishment or ailment is the provision, to cells and organisms,

of the materials necessary (in the form of food) to support life. Many common health

problems can be prevented or alleviated with a healthy diet. The diet of organisms is

what it eats, which is largely determined by the perceived palatability of foods. A

poor diet can have an injurious impact on health, causing deficiency diseases such as

scurvy (According to the report by Linus Pauling Institute, 2011) and kwashiorkor

(According to the report of Medline Plus Medical Encyclopedia, 2011); health-

threatening conditions like obesity (According to the report by National Cancer

Institute, 2011) and metabolic syndrome(According to Med Health Plus report, 2011)

and such common chronic systemic diseases as cardiovascular disease (According to

report by National Diabetes Information Clearing house and Helpguide.org, 2011) and

osteoporosis (Webmd.com , Ods.od.nih.gov and article in The New York Times,

2011).

20

Nutrient

A nutrient is a chemical that an organism needs to live and grow or a substance used

in an organism needs to live and grow or a substance used in an organism’s

metabolism which must be taken in from its environment (Donatelle and Rebecca,

2008 and Whitney et al., 2007). They are used to build and repair tissues, regulate

body processes and are converted to and used as energy.

Nutrients are said to be Organic and Inorganic nutrients. Organic nutrients

include carbohydrates, fats, proteins (or their building blocks, amino acids), and

vitamins. Inorganic chemical compounds such as dietary minerals, water and oxygen

may also be considered as nutrients (Frances sizer et al., 2007). A nutrient is said to

be “Essential” if it must be obtained from an external source, either because the

organism cannot synthesize it or produces insufficient quantities and nutrient is said to

be “Non-essential” if an organism can synthesize it or produces in sufficient

quantities. The effects of nutrients are dose-dependent and shortages are called

deficiencies (Audrey, 1994).

Further, the nutrients are categorized as: Macronutrients (needed in relatively

large amounts) and Micronutrients (needed in small quantities).

The macronutrients include carbohydrates (including fiber), fats, protein and

water.

Calcium, salt (sodium and chloride), magnesium and potassium are sometimes added

to the list of macronutrients because they are required in large quantities compared to

other vitamins and minerals. They are sometimes referred to as the Macrominerals.

The micronutrients are minerals and vitamins.

Dietary minerals are generally trace elements (required in trace amounts), salts

or ions such as copper, iron, cobalt, chromium, iodine, manganese, molybdenum and

zinc. Some of these minerals are essential to human metabolism. Vitamins are organic

compounds essential to the body. They usually act as coenzymes or cofactors for

various proteins in the body.

21

The macronutrients (excluding fiber and water) provide structural material

(amino acids from which proteins are built and lipids from which cell membranes and

some signalling molecules are built) and energy. Some of the structural material can

be used to generate energy internally, in either case it is measured in Joules or

kilocalories.

Carbohydrates and proteins provide 17kJ approximately (4 kcal) of energy per

gram, while fats provide 37 kJ (9 kcal) per gram [Berg et al., 2002] though the net

energy from either depends on such factors as absorption and digestive effort, which

vary substantially from instance to instance. Vitamins, minerals and water do not

provide energy, but are required for other reasons. A third class of class of dietary

material, fiber (i.e., non-digestible material such as cellulose), is also required, for

both mechanical and biochemical reasons.

Multivitamins:

A multivitamin is a preparation intended to supplement a human diet with vitamins,

dietary minerals and other nutritional elements. Such preparations are available in the

form of tablets, capsules, pastilles, powders, liquids and injectable formulations.

Multivitamin supplements are commonly provided in combination with dietary

minerals. A multivitamin is defined as a supplement containing 3 or more vitamins

included at a dose below the tolerable upper level and do not present a risk of adverse

health effects.(ref –search multivitamin Wikipedia). According to The United Nations

authority of food standards - Codex Alimentarius: Guidelines for Vitamin and

Mineral Food Supplements, 2007, multivitamins are recognized as a category of food.

In the mid- 1930’s multivitamins became available in pharmacies and grocery

stores. In 1934 Nutrilite Company introduced the first multivitamin-multimineral

tablets. These supplements were made from natural dried and compressed vegetable

and fruit concentrates. In early 1940’s other brands started to produce synthetic tablets

(Huang, 2006).

Basic commercial multivitamin supplement products often contain the

following ingredients: vitamin C, B1, B2, B3, B6, folic acid (B9), B12, B5

22

(pantothenate), H (biotin), A, E, D3, K1, along with minerals like potassium, zinc,

magnesium, chromium, manganese, molybdenum, beta-carotene and iron.

By supplementing the diet with additional vitamins and minerals,

multivitamins can be a valuable tool for those with dietary imbalances or different

nutritional needs. Individuals who use dietary supplements (including multivitamins)

generally report higher dietary nutrient intakes and healthier diets. Additionally,

adults with a history of prostate and breast cancers are more likely to use dietary and

multivitamin supplements (Cheryl, 2007).

Vitamin deficiencies may result in disease conditions, including goitre, scurvy,

osteoporosis, impaired immune system, disorders of cell metabolism, certain forms of

cancer, symptoms of premature aging and poor psychological health including eating

disorders among many others (Shils, 2005).

While multivitamins can be a valuable tool to correct dietary imbalances,

some risks exists. Deficient or excess levels of vitamins or minerals can also have

serious health consequences. In particular, pregnant women should consult their

doctors before taking any multivitamins: for example, either an excess or deficiency

of vitamin A can cause birth defects (Collins et al., 1999). Long term beta-carotene,

vitamin A, E supplements may shorten life (Randerson, 2008 and Bjelakovic et al.,

2008) with the additional risks.

Essential amino acids:

An essential amino acid or indispensable amino acid is an amino acid that cannot

be synthesized de novo by the organism (usually referring to humans), and therefore

must be supplied in the diet. Although proteins from plant sources tend to have a

relatively low biological value, in comparison to protein from animal sources like

eggs or milk, they are nevertheless "complete" in that they contain at least trace

amounts of all of the amino acids that are essential in human nutrition (Mc Dougall,

2002). Eating various plant foods in combination can provide a protein of higher

biological value (Woolf et al., 2011). Certain native combinations of foods, such as

corn and beans, soybeans and rice, or red beans and rice, contain the essential amino

acids necessary for humans (Ahmed et al., 2009).

23

The amino acids regarded as essential for humans are phenylalanine, valine,

threonine, tryptophan, isoleucine, methionine, leucine, lysine, and histidine (Young,

1994). Additionally, cysteine (or sulphur-containing amino acids), tyrosine (or

aromatic amino acids), and arginine are required by infants and growing children

(Imura and Okada, 1998). Failure to receive even one of these amino acids results in

serious health problems and muscle and bone degradation over time. The body

actually strips them from the muscle and bone structures.

Recommended daily amounts:

Estimating the daily requirement for the indispensable amino acids has proven to be

difficult; these numbers have undergone considerable revision over the last 20 years.

The following table: 2.

Table: 2.

Lists the WHO recommended daily amounts currently in use for essential amino acids

in adult humans, together with their standard one-letter abbreviations

(FAO/WHO/UNU 2007).

Amino acid(s) mg per kg body weight mg per 70 kg mg per 100 kg

(H) Histidine 10 700 1000

(I) Isoleucine 20 1400 2000

(L) Leucine 39 2730 3900

(K) Lysine 30 2100 3000

(M )Methionine

(+ C) Cysteine

10.4 + 4.1 (15 total) 1050 1500

(F) Phenylalanine

(+ Y) Tyrosine

25 (total) 1750 2500

(T) Threonine 15 1050 1500

(W) Tryptophan 4 280 400

(V) Valine 26 1820 2600

24

The recommended daily intakes for children aged three years and older is 10%

to 20% higher than adult levels and those for infants can be as much as 150% higher

in the first year of life.

Use of essential amino acids

List of 9 essential amino acids and the basic functions they perform.

Leucine- Is a branched chain amino acid used for tissue repair after surgery, muscle

mass, blood sugar (diabetics), stress, HGH Human growth hormone, protein

synthesis, bone, skin, weight loss and blood haemoglobin.

Valine- Is a branched chain amino acid used for mental and emotional disorders,

glycogen production, alcohol and drug recovery.

Lysine- Used in absorption and conservation of calcium, bones, concentration,

fertility, Herpes (HSV), cholesterol, hormones, enzymes, Triglycerides, immune

system, skin, collagen and migraines.

Methionine – Used in treatment of Schizophrenia, Parkinsons, heavy metals,

collagen, antioxidant, pancreatitis, endometriosis, liver fat, estrogens’, Arthritis, hair,

skin, nails and depression.

Phenylalanine - Chronic pain, endorphins, alcohol and drug recovery, menstrual

cramps, migraines, Parkinsons, melanoma, Vitiligo and tumours.

Threonine- Tooth enamel, protein balance, immune system, collagen, skin, blood

sugar (diabetics), thymus gland, nervous system, stress, bones, wound healing,

Parkinsons, and Multiple Sclerosis.

Tryptophan- Nerves, anxiety, mental depression, sleep, insomnia, migraines,

fibromyalgia, niacin (B3) production and nicotine withdrawal.

Histidine- Repair tissue, stomach gastric juices, digestion, nerves, ulcers, heavy

metals, red + white blood cells, blood pressure and sexual functioning.

25

Public health initiatives for human beings:

At the 1990 World Summit for Children, the gathered nations identified deficiencies

in three micronutrients-iodine, iron and vitamin A- as being particularly common and

posing public health risks in developing countries[4].Priority programs include

supplementation with vitamin A for children 6-59 months, zinc supplementation as a

treatment for diarrhoeal disease, iron and folate supplementation for women and

children –bearing age, salt iodization, staple food fortification, multiple micronutrient

powders and behaviour centred nutrition education.

In 1997, national vitamin supplementation programming received a boost

when experts met to discuss rapid scale up of supplementation activity and the

micronutrient initiative, with support from the government of Canada, began to ensure

vitamin A supply to UNICEF. Global vitamin A supplementation efforts have

targeted 103 priority country including India. In 1999, 16% of children in these

countries received two annual doses of vitamin A. By 2007, the rate increased to 62%.

Double-fortified salt (DFS) is a public health tool for delivering nutritional

iron. DFS is fortified with both iron and iodine. It was developed by Venkatesh

Mannar, Excecutive Director of the Micronutrient Initiative and University of Toronto

–Professor Levente Diosady, who discovered a process for coating iron particles with

a vegetable fat to prevent the negative interaction of iodine and iron. It was first used

in public programming in 2004. As of September 2010 DFS was being produced only

in the Indian State of Tamil Nadu and distributed through a state school feeding

program.

Salt iodization is the recommended strategy for ensuring adequate human

iodine intake. In1990, less than 20% of households in developing countries were

consuming iodized salt. By 2008 it increased to 72%.

Dietary Reference Intakes:

From 1941 to 1989, the RDA (Dietary Reference Allowance) was the gold standard

used to determine if someone from a particular age and gender group was getting the

nutrients they needed. In the early 1990’s the Food and Nutrition Board decided these

26

benchmarks needed an overhaul. Out of those revision came a whole new way to

evaluate nutrients, the DRI’s (Dietary Reference Intakes).

Between 1997 and 2004 the values were written for 46 different nutrient

substances, including vitamins, many more minerals than were included in the RDA,

electrolytes, water and macronutrients such as fiber, carbohydrates, fatty acids,

cholesterol and amino acids.

DRI’s actually include four different reference values:

The RDA, which is the daily intake of a nutrient sufficient to meet the needs

of almost all healthy people in an age and gender group.

The AL or Adequate Intake, which is used if the RDA cannot be established.

This is based on looking at how much of a nutrient healthy people actually eat.

The UL is the Tolerable Upper Intake Level, or the amount of a healthy

person can eat without any risk of toxicity.

The EAR, or Estimated Average Requirement, is the amount of a nutrient that

is estimated to meet the requirement of half of all healthy individuals in the

populations.

Of these four reference values, the RDA, AL and UL are most use to individuals

who want to know a safe and healthy level of nutrient intake. The EAR is mainly of

use to people who are planning diets for large population groups or developing new

foods.

The DRI’s of the vitamins, micro and macro elements pertaining to age and

gender are shown in the table 3, 4 and 5.

27

Table: 3. DRI (Dietary reference intake): Recommended intake for vitamins per day.

AGE Vit

A

(µg)

Thia

min

(mg)

Ribof

lavin

(mg)

Niac

in

(mg)

Vit

B6

(mg)

Fola

te

(µg)

Vit

B12

(µg)

Panto

thenic

acid

(mg)

Biotin

(µg)

Chol

ine

(mg)

Vit

C

(mg)

Vit

D

(µg)

Vit

E

(mg)

Vit

K

(µg)

INFANTS

0-6 Months 400 0.2 0.3 2 0.1 65 0.4 1.7 5 125 40 5 4 2.0

7-12 Months 500 0.3 0.4 4 0.3 80 0.5 1.8 6 150 50 5 5 2.5

CHILDREN

1-3 300 0.5 0.5 6 0.5 150 0.9 2 8 200 15 5 6 30

4-8 400 0.6 0.6 8 0.6 200 1.2 3 12 250 25 5 7 55

MALES

9-13 600 0.9 0.9 12 1.0 300 1.8 4 20 375 45 5 11 60

14-18 900 1.2 1.3 16 1.3 400 2.4 5 25 550 75 5 15 75

19-30 900 1.2 1.3 16 1.3 400 2.4 5 30 550 90 5 15 120

31-50 900 1.2 1.3 16 1.3 400 2.4 5 30 550 90 5 15 120

51-70 900 1.2 1.3 16 1.7 400 2.4 5 30 550 90 10 15 120

70+ 900 1.2 1.3 16 1.7 400 2.4 5 30 550 90 15 15 120

FEMALES

9-13 600 0.9 0.9 12 1.0 300 1.8 4 20 375 45 5 11 60

14-18 700 1.0 1.0 14 1.2 400 2.4 5 25 400 65 5 15 75

19-30 700 1.1 1.1 14 1.3 400 2.4 5 30 425 75 5 15 90

31-50 700 1.1 1.1 14 1.3 400 2.4 5 30 425 75 5 15 90

51-70 700 1.1 1.1 14 1.5 400 2.4 5 30 425 75 10 15 90

70+ 700 1.1 1.1 14 1.5 400 2.4 5 30 425 75 15 15 90

PREGNANT

14-18 750 1.4 1.4 18 1.9 600 2.6 6 30 450 80 5 15 75

19-30 770 1.4 1.4 18 1.9 600 2.6 6 30 450 85 5 15 90

31-50 770 1.4 1.4 18 1.9 600 2.6 6 30 450 85 5 15 90

LACTATION

14-18 120

0

1.4 1.6 17 2.0 500 2.8 7 35 550 115 5 19 75

19-30 130

0

1.4 1.6 17 2.0 500 2.8 7 35 550 120 5 19 90

31-50 130

0

1.4 1.6 17 2.0 500 2.8 7 35 550 120 5 19 90

28

Table: 4. DRI (Dietary reference intake): Recommended intake for elements per day.

AGE Calcium

(mg)

Iodine

(µg)

Iron

(mg)

Magnesium

(mg)

Manganese

(mg)

Molyb

denum

(µg)

Phosph

orus

(mg)

Zinc

(mg)

Potassi

um

(gm)

Sodium

(gm)

INFANTS

0-6 Months 210 110 0.27 30 0.003 2 100 2 0.4 0.12

7-12 Months 270 130 11 75 0.6 3 275 3 0.7 0.37

CHILDREN

1-3 500 90 7 80 1.2 17 460 3 3.0 1.0

4-8 800 90 10 130 1.5 22 500 5 3.8 1.2

MALES

9-13 1300 120 8 240 1.9 34 1250 8 4.5 1.5

14-18 1300 150 11 410 2.2 43 1250 11 4.7 1.5

19-30 1000 150 8 400 2.3 45 700 11 4.7 1.5

31-50 1000 150 8 420 2.3 45 700 11 4.7 1.5

51-70 1200 150 8 420 2.3 45 700 11 4.7 1.3

70+ 1200 150 8 420 2.3 45 700 11 4.7 1.2

FEMALES

9-13 1300 120 8 240 1.6 34 1250 8 4.5 1.5

14-18 1300 150 15 360 1.6 43 1250 9 4.7 1.5

19-30 1000 150 18 310 1.8 45 700 8 4.7 1.5

31-50 1000 150 18 320 1.8 45 700 8 4.7 1.5

51-70 1200 150 8 320 1.8 45 700 8 4.7 1.3

70+ 1200 150 8 320 1.8 45 700 8 4.7 1.2

PREGNANT

14-18 1300 220 27 400 2.0 50 1250 12 4.7 1.5

19-30 1000 220 27 350 2.0 50 700 11 4.7 1.5

31-50 1000 220 27 360 2.0 50 700 11 4.7 1.5

LACTATION

14-18 1300 290 10 360 2.6 50 1250 13 5.1 1.5

19-30 1000 290 9 310 2.6 50 700 12 5.1 1.5

31-50 1000 290 9 320 2.6 50 700 12 5.1 1.5

29

Table: 5. DRI (Dietary Reference Intake): Recommended intake for

macronutrients per day

AGE Total water

(litres)

Carbohydrate

(grams)

Total fiber

(grams)

Fat

(grams)

Protein

(grams)

INFANTS

0-6 Months 0.7 60 ND 31 9.1

7-12 Months 0.8 95 ND 30 11

CHILDREN

1-3 1.3 130 19 ND 13

4-8 1.7 130 25 ND 19

MALES

9-13 2.4 130 31 ND 34

14-18 3.3 130 38 ND 52

19-30 3.7 130 38 ND 56

31-50 3.7 130 38 ND 56

51-70 3.7 130 30 ND 56

70+ 3.7 130 30 ND 56

FEMALES

9-13 2.1 130 26 ND 34

14-18 2.3 130 26 ND 46

19-30 2.7 130 25 ND 46

31-50 2.7 130 25 ND 46

51-70 2.7 130 21 ND 46

70+ 2.7 130 21 ND 46

PREGNANT

14-18 3.0 175 28 ND 71

19-30 3.0 175 28 ND 71

31-50 3.0 175 28 ND 71

LACTATION

14-18 3.8 210 29 ND 71

19-30 3.8 210 29 ND 71

31-50 3.8 210 29 ND 71

ND: Not detected.

30

Sauropus androgynous, apart from being popularised as multivitamin plant and

consumed as leafy green vegetable it is regionally and commercially threatened and

nearly banned for consumption causing Bronchiolitis Obliterans due to the presence

of alkaloid papaverine. But now the world is obsessed with two words-

MALNUTRITION AND OBSESITY.

“Good nutrition” early in life is a key input for sustainable and equitable

economic growth. With persistently high levels of child under nutrition, vital

opportunities to save millions of lives are being lost and many more children are not

growing to their full potential. Malnutrition is considered as a “Matter of shame” for

any economically booming country and concerned governments are initiating vitamin

supplemented programmes. Another problem which goes parallel with malnutrition is

overeating. Overeating leads to obesity, in turn leading to many a disease. Hence

people have become much calorie conscious and are fascinated towards being slim.

Thus some where the nutritional requirement in their daily diet is lost. At this

juncture S. androgynous proves absolute boon to the present world as it is a

perfect blend of being nutritious, slimming with other medicinal properties when

taken in restricted quantity. Thus there is a much need to conserve this

“WONDER PLANT”.

Though conventional method of conservation by vegetative propagation easily

satisfies the multiplication of this plant but takes a back seat when comes to

commercial acceptance of the same. Hence conservation strategy here should not

solely highlight on just multiplying it but instead it has to be highlighted on

reintroduction of this plant in better and safer way for human consumption. But

this is negligibly very low or nil conventionally because recombination breeding for

occurrence of genetic variation of new variety is handicapped by negligible amount of

fruit set, poor seed longevity and short viable period of seed. Hence an attempt for in

vitro propagation via, Organogenesis and Somatic embryogenesis in combination with

mutagenesis was done which will be useful in producing innovative safer variety of

Sauropus androgynous.

31

OBJECTIVES

The presented study was designed with the following objectives:

1. To regenerate Sauropus androgynous using shoot tip, leaf and nodal explants

through:

a. Callus cultures.

b. Organogenesis- Direct organogenesis and indirect organogenesis.

c. Somatic embryogenesis - Primary somatic embryogenesis, Secondary

somatic embryogenesis and Tertiary somatic embryogenesis.

2. To facilitate rapid growth in the regenerates by:

a. Preparing modified medium by varying the concentrations of vitamins.

b. Supplementing medium with different amino acids.

3. To induce variability in the regenerates by exposing/subjecting explants to

varying doses of mutagens like:

a. Physical mutagen – Ethyl Methyl Sulphonate (EMS).

b. Chemical mutagen – Gamma irradiation (� rays).

4. To establish the regenerated plants in the field.

5. To estimate the Papaverine (alkaloid) content by HPLC studies in normal and

regenerated plants.

6. To estimate nutritional and phytochemical contents in normal and regenerated

leaves based on papaverine content which includes:

A. To estimate the chlorophyll content.

B. To estimate primary metabolites like carbohydrates, proteins and reducing

sugars.

C. To estimate total ash content, fibre, fat, moisture, pH, acid value and

calorific value/energy.

D. To estimate Vitamins (A, B, C, D, E and K) in normal and regenerated

plants.

E. To analyze and estimate the essential amino acids content.

H. To analyze and estimate mineral content:

a. Macro minerals- Calcium, Iron, Magnesium, Potassium, Phosphorus and

Sodium. Sodium.

b. Micro minerals- Molybdenum, Zinc and Copper.