INTRODUCTION:

Herbal medicines are the synthesis of therapeutic experiences of generations of practicing physicians of indigenous systems of medicine for over hundreds of years while nutraceuticals are nutritionally or medicinally enhanced foods with health benefits of recent origin and marketed in developed countries. In 1991, WHO developed guidelines for the assessment of herbal medicine.The earliest recorded evidence of their use in Indian, Chinese,Egyptian, Greek, Roman and Syrian texts dates back to about5000 years. The classical Indian texts include Rigveda, Atherveda,Charak Samhita and Sushruta Samhita. The herbalMedicines/traditional medicaments have, therefore, been derivedfrom rich traditions of ancient civilizations and scientific heritage.

WHY??

Herbal medicines are being used by about 80% of the worldpopulation primarily in the developing countries for primaryhealth care. They have stood the test of time for their safety,efficacy, cultural acceptability and lesser side effects. Ancient literature also mentions herbal medicines for age-related diseases namely memory loss, osteoporosis, diabetic wounds, immune and liver disorders, etc. for which no modern medicine or only palliative therapy is available. These drugs are made from renewable resources of raw materials by ecofriendly processes and will bring economic prosperity to the masses growing these raw materials.

WORLD MARKET

As per available records, the herbal medicine market in 1991 in the countries of the European Union was about $ 6 billion (may be over $ 20 billion now), with Germany accounting for $ 3 billion, France $ 1.6 billion and Italy $ 0.6 billion. Incidentally in Germany and France, herbal extracts are sold as prescription drugs and are covered by national health insurance. In 1996, the US herbal medicine market was about $ 4 billion and with the current growth rate may be more than double by the turn of century. Thus a reasonable guesstimate for current herbal medicine market worldwide may be around $ 30–60 billion. The Indian herbal drug market is about $ one billion and the export of herbal crude extracts is about $ 80 million. Germany holds the lead and has published individual monographs on therapeutic benefits of more than 300 herbs. In developing countries, China has compiled/generated data on over 800 medicinal plants and exports large amounts of herbal drugs. India has prepared only a few monographs and its exports are dismal.

Herbal medicine scenario in IndiaThe turnover of herbal medicines in India as over-the-counterproducts, ethical and classical formulations and home remedies of Ayurveda, Unani and Siddha systems of medicine is about $ 1billion with a meagre export of about $ 80 million. Psyllium seeds and husk, castor oil and opium extract alone account for 60% of the exports. 80% of the exports to developed countries are of crude drugs and not finished formulations leading to low revenue for the country. Thus the export of herbal medicines from India is negligible despite the fact that the country has a rich traditional knowledge and heritage of herbal medicine. India is one of the 12 mega biodiversity centres having over 45,000 plant species. Its diversity is unmatched due to the presence of 16 different agro climatic zones, 10 vegetative zones and 15 biotic provinces. The country has 15,000–18,000flowering plants, 23,000 fungi, 2500 algae, 1600 lichens, 1800bryophytes and 30 million micro-organisms. India also has equivalent to 3/4 of its land exclusive economic zone in the ocean harbouring a large variety of flora and fauna, many of them with therapeutic properties. About 1500 plants with medicinal uses are mentioned in ancient texts and around 800 plants have been used in traditional medicine. The major traditional sector pharmas, namely Himalaya, Zandu,Dabur, Hamdard, Maharishi, etc. and modern sector pharmas,namely Ranbaxy, Lupin, Allembic, etc. are standardizing theirherbal formulations by chromatography techniques likeTLC/HPLC finger printing, etc. There are about 7000 firms in the small-scale sector manufacturing traditional medicines with or without standardization.

REVIEW OF LITERATURE:

Macrothelypteris torresiana

Conservation status

Apparently Secure (NatureServe)

Scientific classification

Kingdom: Plantae

Division: Polypodiophyta

Class: Polypodiopsida

Order: Blechnales

Family: Thelypteridaceae

Genus: Macrothelypteris

Species: M. torresiana

Binomial name

Macrothelypteris torresiana

(Gaudich.)

Synonyms

Thelypteris torresiana

Macrothelypteris torresiana is a species of fern which is native to tropical and subtropical

Africa and Asia. It has been introduced into other areas, including large parts of North and

South America.[1][2]

The species is assigned to different genera depending on how the family Thelypteridaceae

is classified. If the entire family is classified in Thelypteris, it is Thelypteris torresiana.

However, molecular data shows an affinity between a group of species which are

sometimes classified in Macrothelypteris,Phegopteris, and allied genera,[3] so it is common

to remove this species from Thelypteris and classify it as Macrothelypteris torresiana.

M. torresiana contains flavanoids which have been investigated for possible medicinal value

Description

Macrothelypteris torresiana is a robust fern with

a short creeping rhizome. Fronds are

three-pinnate, deeply divided, to 1.2 m long, with

a stalk 0.5 m. Sori are circular but not terminal

on the veins.

Fertile: September.

AFRICA Southern Africa: South Africa - KwaZulu-NatalWestern Indian Ocean: Madagascar

ASIA-TEMPERATE Eastern Asia: Japan - Honshu, Kyushu [s. & w.], Ryukyu Islands, Shikoku [s.]; Taiwan

ASIA-TROPICAL Indo-China: Indochina; ThailandMalesia: Indonesia; Malaysia

AUSTRALASIA Australia: Australia - Queensland [e.]

New Zealand: New Zealand PACIFIC 

North-Central Pacific: United States - HawaiiSouth-Central Pacific: French PolynesiaSouthwestern Pacific: Samoa

it grows in open

places usually near water

FINDINGS TILL NOW:

Two new flavone derivatives (1 and 2) were isolated from the aerial parts of Macrothelypteris

torresiana, along with four known flavonoids: protoapigenin, apigenin, kaempferol and quercetin. The

structures were determined on the basis of spectroscopic data. Compound 1 showed weak cytotoxic

activity against human tumour cell lines HepG2, MCF7 and K562.

Preliminary Phytochemical Screening

MATERIALS AND METHODS

Collection of Plant Materials

Aerial parts of Macrothelypteris torresiana (Gaudich.) Ching was obtained from a domestic stand in Kadiri, Anantapur District, Andhra Pradesh State, India. The plant materials were transported in polythene bags to the Research Laboratory of Pharmacy, GITAM UNIVERSITY, VSKP, AP, INDIA. where the study was carried out.

Processing of Plant Materials

The aerial parts were washed in running water and cut into small bits to facilitate drying. The pieces of plant material were dried in room temperature for 48 hours.The dried plant materials (aerial parts) was taken separately and ground using an electric blender to obtain a fine powder. The powder was further passed through a 2mm sieve to obtain finer particles. The powdered samples were stored in a clean glassware container until needed for analysis.

Solvent Extraction Procedures

5g portions of powdered plant materials were each separately dispersed in 50ml of each water, 70% ethanol, acetone, methanol and hexane. The solution was left to stand at room temperature for 24hrs and was filtered with Whatman No. 1 filter paper. The filtrate was used for the phytochemical screening using the following tests.

Methods of Extraction of Medicinal Plants

Maceration

In this process, the whole or coarsely powdered crude drug is placed in a stoppered container with the solvent and allowed to stand at room temperature for a period of at least 3 days with frequent agitation until the soluble matter has dissolved. The mixture then is strained, the marc (the damp solid material) is pressed, and the combined liquids are clarified by filtration or decantation after standing.

circulatory extraction.

InfusionFresh infusions are prepared by macerating the crude drug for a short period of time with cold or boiling water. These are dilute solutions of the readily soluble constituents of crude drugs.

DigestionThis is a form of maceration in which gentle heat is used during the process of extraction. It is used when moderately elevated temperature is not objectionable. The solvent efficiency ofthe menstrum is thereby increased.

DecoctionIn this process, the crude drug is boiled in a specified volume of water for a defined time; it is then cooled and strained or filtered. This procedure is suitable for extracting water-soluble, heatstable constituents. This process is typically used in preparation of Ayurvedic extracts called “quath” or “kawath”. The starting ratio of crude drug to water is fixed, e.g. 1:4 or 1:16; the volume is then brought down to one-fourth its original volume by boiling during the extraction procedure. Then, the concentrated extract is filtered and used as such or processed further.

PercolationThis is the procedure used most frequently to extract active ingredients in the preparation of tinctures and fluid extracts. A percolator (a narrow, cone-shaped vessel open at both ends) isgenerally used. The solid ingredients are moistened with an appropriate amount of the specified menstruum and allowed to stand for approximately 4 h in a well closed container, after which the mass is packed and the top of the percolator is closed. Additional menstruum is added to form a shallow layer above the mass, and the mixture is allowed to macerate in the closed percolator for 24 h. The outlet of the percolator then is opened and the liquid contained therein is allowed to drip slowly. Additional menstruum is added as required, until the percolate measures about three-quarters of the required volume of the finished product. The marc is then pressed and the expressed liquid is added to the percolate. Sufficient menstruum is added to produce the required volume, and the mixed liquid is clarified by filtration or by standing followed by decanting.

Hot Continuous Extraction (Soxhlet)In this method, the finely ground crude drug is placed in a porous bag or “thimble” made of strong filter paper, which is placed in chamber E of the Soxhlet apparatus (Figure 2). Theextracting solvent in flask A is heated, and its vapors condense in condenser D. The condensed extractant drips into the thimble containing the crude drug, and extracts it by contact. When the

level of liquid in chamber E rises to the top of siphon tube C, the liquid contents of chamber E siphon into fl ask A. This process is continuous and is carried out until a drop of solvent from thesiphon tube does not leave residue when evaporated. The advantage of this method, compared to previously described methods, is that large amounts of drug can be extracted with a much smaller quantity of solvent. This effects tremendous economy in terms of time, energy and consequently financial inputs. At small scale, it is employed as a batch process only, but it becomes much more economical and viable when converted into a continuous extraction procedure on medium or large scale.

Aqueous Alcoholic Extraction by FermentationSome medicinal preparations of Ayurveda (like asava and arista) adopt the technique of fermentation for extracting the active principles. The extraction procedure involves soaking thecrude drug, in the form of either a powder or a decoction (kasaya), for a specified period of time, during which it undergoes fermentation and generates alcohol in situ; this facilitates the extraction of the active constituents contained in the plant material. The alcohol thus generated also serves as a preservative. If the fermentation is to be carried out in an earthen vessel, it should not be new: water should first be boiled in the vessel. In large-scale manufacture, wooden vats, porcelain jars or metal vessels are used in place of earthen vessels. Some examples of such preparations are karpurasava, kanakasava, dasmularista. In Ayurveda, this method is not yet standardized but, with the extraordinarily high degree of advancement in fermentation technology, it should not be difficult to standardize this technique of extraction for the production of herbal drug extracts.Counter-current ExtractionIn counter-current extraction (CCE), wet raw material is pulverized using toothed disc disintegrators to produce a fine slurry. In this process, the material to be extracted is moved inone direction (generally in the form of a fine slurry) within a cylindrical extractor where it comes in contact with extraction solvent. The further the starting material moves, the more concentrated the extract becomes. Complete extraction is thus possible when the quantities of solvent and material and their flow rates are optimized. The process is highly efficient,

requiring little time and posing no risk from high temperature. Finally, sufficiently concentrated extract comes out at one end of the extractor while the marc (practically free of visible solvent) falls out from the other end.This extraction process has significant advantages:

A unit quantity of the plant material can be extracted with much smaller volume of solvent as compared to other methods like maceration, decoction, percolation.CCE is commonly done at room temperature, which spares the thermolabile constituents fromexposure to heat which is employed in most other techniques.As the pulverization of the drug is done under wet conditions, the heat generated during comminution is neutralized by water. This again spares the thermolabile constituents from exposure to heat.The extraction procedure has been rated to be more efficient and effective than continuous hot extraction.

Ultrasound Extraction (Sonication)The procedure involves the use of ultrasound with frequencies ranging from 20 kHz to 2000 kHz; this increases the permeability of cell walls and produces cavitation. Although theprocess is useful in some cases, like extraction of rauwolfia root, its large-scale application is limited due to the higher costs. One disadvantage of the procedure is the occasional but knowndeleterious effect of ultrasound energy (more than 20 kHz) on the active constituents of medicinal plants through formation of free radicals and consequently undesirable changes in the

drug molecules.

Supercritical Fluid ExtractionSupercritical fluid extraction (SFE) is an alternative sample preparation method with general goals of reduced use of organic solvents and increased sample throughput. The factors toconsider include temperature, pressure, sample volume, analyte collection, modifier (cosolvent) addition, flow and pressure control, and restrictors. Generally, cylindrical extraction vessels are used for SFE and their performance is good beyond any doubt. The collection of the extracted analyte following SFE is another important step:Significant analyte loss can occur during this step, leading the analyst to believe that the actual efficiency was poor.There are many advantages to the use of CO2 as the extracting fluid. In addition to its favorable physical properties, carbon dioxide is inexpensive, safe and abundant. But while carbon dioxide is the preferred fluid for SFE, it possesses several polarity limitations. Solvent polarity is important when extracting polar solutes and when strong analyte-matrix interactions are present. Organic solvents are frequently added to the carbon dioxide extracting fluid to alleviate the polarity

limitations. Of late, instead of carbon dioxide, argon is being used because it is inexpensive and more inert. The component recovery rates generally increase with increasing pressure or temperature: the highest recovery rates in case of argon are obtained at 500 atm and 150° C.The extraction procedure possesses distinct advantages:

The extraction of constituents at low temperature, which strictly avoids damage from heat and some organic solvents.No solvent residues.Environmentally friendly extraction procedure.

Phytochemical Screening

Test for Alkaloids (Wagner’s reagent)

A fraction of extract was treated with 3-5drops of Wagner’s reagent [1.27g of iodine and 2g of potassium iodide in

100ml of water] and observed for the formation of reddish brown precipitate (or colouration).

Test for Carbohydrates (Molisch’s test)

Few drops of Molisch’s reagent were added to 2ml portion of the various extracts. This was followed by addition of

2ml of conc. H2SO4 down the side of the test tube. The mixture was then allowed to stand for two-three minutes.

Formation of a red or dull violet colour at the interphase of the two layers was a positive test.

Test for Cardiac glycosides (Keller Kelliani’s test)

5ml of each extract was treated with 2ml of glacial acetic acid in a test tube and a drop of ferric chloride solution was added to it. This was carefully underlayed with 1ml concentrated sulphuric acid. A brown ring at the interface indicated the presence of deoxysugar characteristic of cardenolides. A violet ring may appear below the ring while in the acetic acid layer, a greenish ring may form.

Test for Flavonoids (Alkaline reagent test) 2ml of extracts was treated with few drops of 20% sodium

hydroxide solution. Formation of intense yellow colour, which becomes colourless on addition of dilute hydrochloric acid, indicates the presence of flavonoids.

IN VIVO METHODS FOR TESTING CENTRAL ANALGESIC ACTIVITY

HAFFNER’s tail clip method Radiant heat method Hot plate method . Tail immersion test Electrical stimulation of the tail Grid shock test Tooth pulp stimulation Monkey shock titration test Formalin test in rats

ANTI PYRETIC ACTIVITY

Anti-pyretic testing in rats Anti-pyretic testing in rabbits

IN VIVO METHODS FOR ANTI-INFLAMMATORY ACTIVITY METHODS FOR TESTING ACUTE AND SUBACUTE INFLAMMATION

Ultraviolet erythema in guinea pigsVascular permeabilityInhibition of leukocyte adhesion to rat mesenteric venules in vivo Oxazolone-induced ear edema in mice Croton-oil ear edema in rats and mice Paw edema Pleurisy test Granuloma pouch technique Urate-induced synovitis Methods for testing the proliferative phase (granuloma formation) Cotton wool granuloma Sponge implantation technique Glass rod granuloma Side effects of anti-inflammatory compounds General considerations Ulcerogenic effect in rats Measurement of gastric mucosal damage by intragastric insulin