WO2013183064A2

WO2013183064A2 - Extraction of phytochemicals from trichilia connaroides

Info

Publication number: WO2013183064A2
Application number: PCT/IN2013/000350
Authority: WO
Inventors: Ramaswamy Rajendran; Kamala Rajendran; Ashok PURNIMA
Original assignee: Ramaswamy Rajendran; Kamala Rajendran
Priority date: 2012-06-06
Filing date: 2013-06-03
Publication date: 2013-12-12
Also published as: WO2013183064A3

Abstract

In the extraction of TC(Trichilia connaroides) by solvent(s), the former is reacted with acidic plant matter such as, for example, amla fruit matter(Emblica officinalis) which converts some of the insoluble principles therein into soluble forms, thus enhancing the extraction range and yield. In extraction by water, the adoption of amla fruit matter increases the TC principles yield from about 10% to about 12% by wt., of the saponins from about 8.2% to about 12.3% by wt. and of the terpenes from about 1.1% to about 3.9% by wt. The corresponding yield figures in extraction by aq. ethanol, respectively with and without the adoption of amla fruit matter are 14%, 15.6% and 5.5% by wt. and 8%, 11.1% and 4.1% by wt. for the three sets of compounds respectively. With both solvents, the adoption of amla increases the yields, said increases being greater in the case of aq. ethanol.

Description

'Extraction of phytochemicals from Trichilia connaroides' Introduction to the Invention

^' 5

This invention relates to a process of extraction of plant matter and more specifically to a process of extraction of medicinal principles, vitamins and minerals and other phytochemical constituents of Trichilia connaroides, wherein a first plant matter comprising one or more parts of Trichilia connaroides plant is extracted with a solvent, or solvent mixture, to yield an extract solution, and 0 characterised in that said first plant matter is treated with a second plant matter(s) containing acid(s)/acidic compound(s) such as to, at least, convert one or more of the solvent-insoluble constituents of said first plant matter into form(s) that are soluble in said solvent or solvent mixture, said treatment being carried out, prior to, or during said extraction or both. 5 Background of the Invention and the Objects and Synopsis of the Invention

The genus, Trichilia is known to consist of about 40 species. These species comprise biologically and pharmacologically active constituents. The genus belongs to the family, Meliaceae that includes numerous species which contain insecticidal and pesticidal constituents.

0

Trichilia connaroides(Wight and Arn.) Bentv. is one of the few plants of the Trichilia group, that is found in India. It is found to grow in moist forests in the sub-Himalayan and the Western Ghats regions. It is also found in other tropical regions in the country. Outside India, some of the other regions where Trichilia species are found are Nepal, South China, Malaysia and5 Indonesia.

Other names by which it is known are Zanthoxylum connaroides(Wight and Arn.) Bentv.and Heynea trijuga(Roxb. ex sims). Common names by which it is known by people in India are: Karai or Karaivilangu in Tamil; Korakudi and Kuravatti in Malayalam: Kora in Kannada; and0 Gundira in Marathi. The other plant materials mentioned in this specification are also found in India.

It is a rich source of bioactive compounds such as limonoids, compounds resulting from the degradation of the limonoids, steroids, triterpenoids and others. It is a good source of the5 tetracyclic triterpenes (Heynic acid I and Π), tetranorterpenoids (Trijugin A and Trijugin B), Trijugin B acetate, and pentanortriterpenoids with a novel carbon skeleton Trijugin C[l-4]. These compounds of interest occur in the leaves, flowers, roots, pericarp, twigs and fruits of the plant[l-4, 6-8] and possibly also in other parts of the plant such as the bark, stems, rhizomes etc.

The presence of some sesquiterpenes in the solvents extracts of different parts of the genus Trichilia has been reported by Agarwal G[9]. Some sesquiterpenoids have also been found in the methanol extract of some Trichilia genus plants.

A certain amount of traditional medicinal folklore is associated with Trichilia group of plants. There are several references to its medicinal properties in traditional medicine in India and the medicinal activities of the plant have been reported in several publications in India. In Chinese traditional medicine the roots of T. connaroides are reported to be used in the treatment of arthritis, pharingitis, tonsilitis and other ailments.[4].

Medicinal, insecticidal and pesticidal properties of Trichilia connaroides extracts that have been actually investigated and reported in the literature are:

(i) Antihyperlipidemic effect in hypercholesterolemic rats; [5]

(ii) Antihyperhomocysteinemic and antihyperlipidemic effects in methionine-induced

hyperhomocysteinemic animals;(12)

(iii) Growth-regulatory activity against the Bihar hairy caterpillar, Spilosoma

obliquafLepidoptera: Arctiidae;(13)

(iv) Hypocholesterolemic and antihypercholesterolemic activity of extracts of Trichilia

connaroides on rats;(14) and

(v) Analgesic and anti-inflammatory activity of the chloroform extract of Trichilia

connaroides; (11).

Conventionally, the, food, medicinal and nutritional constituents of plant matter are extracted by contacting the plant matter with a solvent, the choice of the solvent being one that dissolves the desired constituent or the group of constituents of interest. A solvent is chosen that dissolves as many of the desired constituents as possible.

Such first extraction of the plant matter by a first solvent is referred to as a primary extraction and the said first solvent as the primary solvent. The extract obtained is referred to as the primary extract. It is a dilute extract which may be concentrated to give a solid or semi-solid product or a concentrated solution product, which product forms are also referred to as primary extracts. Primary extracts are also referred to as crude extracts in the art. A primary extract contains a plurality of the constituents of the original plant matter, some of which may be of interest and some not.

For the constituents of interest that are substantially insoluble in said first solvent and therefore not extractable by it are then extracted by means of a second solvent in a second extraction operation. The second solvent is contacted with either the spent plant matter coming from the first primary extraction or other, or alternatively with fresh plant matter.

Such second extracts and extractions are also referred to as primary or crude extracts and extractions respectively. This is because they also comprise a plurality of the said constituents in solution therein.

To obtain extraction of substantially all of the desired constituents from a plant matter, generally a minimum of two primary extractions are necessary. For example one may be by ethanol or aq. ethanol and another by hexane to extract out the fat constituents. The term 'constituents' is intended herein to mean generally any of the compounds contained in the plant matter including the food, medicinal and nutritional compounds, vitamins and minerals, anti-oxidants and like compounds therein that are relevant in health, food, nutraceuticals, nutrition, medicine, aging and rejuvenation, recovery and other considerations. Said constituents are also referred to as 'nutrients' herein in the interests of conciseness.

Such primary extracts are optionally further treated in the art to achieve separation of said constituents into groups of compounds or into individual compounds. This is usually done by fractional extraction procedures or partitioning procedures, employing one or more secondary solvents. Chromatographic methods are also used for said separations. Often combinations of chromatographic methods and the said fractionation or partitioning procedures are adopted. The separated compounds are admixed together to make different formulations of the nutrients. Different procedures and extraction solvents have been reported in the prior art for the extraction of Trichilia connaroides extraction These are briefly described hereinbelow. The term 'Trichilia connaroides' has been abbreviated to the initials 'TC herein, in the interests of conciseness. In the investigation of Prasanna GS et al [12], TC leaves were procured and shade-dried. The shade dried leaves were coarse powdered. The leaf powder was defatted with n-hexane by leaving the powder with the solvent overnight with occasional shaking agitation. The plant matter was then successively extracted with chloroform and methanol in a soxhlet extractor. The CETC extract(chloroform extract) and the METC extract(methanol extract) were stored in airtight containers in a cool, dry place^away from sunlight till further use.

Similar procedure has been adopted in Prasanna GS and Purnima A[5], which work is also on the medicinal activity of CETC and METC extracts of TC. The procedure in Purnima A et al, [10], is also similar. The marc(spent plant matter) remaining from the chloroform extraction is extracted twice with methanol.

The procedure adopted by Purnima A, et al[l 1] is also similar. It is an investigation of the analgesic and anti-inflammatory activity of T. connaroides. The defatted plant matter is then extracted with chloroform, distilled and the last traces of the solvent removed by vacuum. The coarse powder is successively extracted with a set of solvents in the order of increasing polarity to isolate the different phytoconstituents. The solvents adopted were: hexane, benzene, chloroform, and mixture of chloroform and ethyl acetate. They report that the presence of a flavonoid, a glycoside, steroids, saponins and phenolic compounds was established. In the extraction procedure adopted by Tandon S, et al [13], TC leaves were collected

and shade-dried. The dried leaves were powdered and the powder extracted separately with acetone or n-hexane in a Soxhlet apparatus. The extracts were concentrated in a flash evaporator under reduced pressure at 30 + 5 C. The crude extracts were diluted with the respective solvents and used in the bioassays.

Said prior art processes extract a selected nutrient solute or a group of solutes. The adoption of the process of the invention allows one to select any group of the said nutrient solutes and obtain them in a single extract through a single primary extract operation unlike the prior art processes. This can be done by suitable selection of the one or more plant species comprising said second plant matter(s).

It may be noted that the extract obtained by the process of the invention is a ready formulation comprising the desired constituents in a ready-mixed form. This demonstrates a major advantage of the process of this invention as it eliminates a number of processing steps required in arriving at such formulations, in the prior art. In particular, the process of the invention does away with the necessity of multiple extractions. As mentioned, at least two said primary extractions are generally necessary to extract out a set of said plant constituents of interest. Reducing the number of extraction operations required can considerably bring down the capital and manufacturing costs besides offering other cost and processing benefits. It would be most advantageous if all the constituents of interest could be extracted by a single solvent in a single extraction operation.

Many of these factors, principles and compounds coming under the purview of the term

'nutrients' and 'constituents' exhibit their medicinal, nutritional and other properties of interest in more than one form. What is therefore required is to convert the nutrient(s) that are insoluble in the adopted solvent into form(s) that are soluble therein. This approach makes it possible to get all, or substantially all of the nutrients of interest together into a single extract by means of a single extraction operation.

This novel approach has been adopted in the co-pending Indian application for patent No.

2677/CHE/2011 of 4^th Aug. 2011 by the present applicants/inventors. This invention deals with the extraction of plant matters by water. Said specification is adopted in the present specification by reference.

Said Indian application 2677 did not contemplate ethanol solvent. As a solvent, ethanol has some unique advantages that are not available in the use of water or other solvents. Aqueous ethanols have also not been contemplated in said application. The adoption of an aq. ethanol as solvent offers the combination of advantages associated with water and ethanol as solvents. The present invention has adopted said aqueous ethanols as solvents together with said acidic components) for the extraction of T. connaroides plant matter. The present invention has also adopted and investigated the process of extraction T. connaroides by water solvent along with the adoption of said acidic second plant matter(s).

This invention has observed unexpected and surprising advantages in the both the processes, the water-based process using said acidic second plant matter(s) and the aq.ethanol based process also using the said acidic second plant matter(s). The aq. ethanol based process turns out to show more advantages over the water based process with regard to the yield and the spread of the extractable constituents, and others. This invention has adopted the idea presented in said application 2677 of reacting the constituents that are insoluble in the adopted solvent such as to convert them into form(s) that are soluble therein. This widens the range of extractable constituents. This invention has also adopted the idea given in said application 2677 of applying acid(s) and/or acidic compounds for said conversion.

The present invention, like the application 2677, calls for preferably using plant matter(s) containing one or more acids/acidic compounds for changing said insoluble constituents) into soluble form(s). The said acidic plant matter is referred to therein and herein as the second plant matter(s). Said second plant matter(s) may contain free acid(s) or acidic component(s) or both. Thus, in the further specification herein the term 'acidic plant matter' is intended to mean that the matter contains either free acid(s) or acidic compound(s) or both. The plant matter being extracted is referred to as the first plant matter therein and herein.

It will be observed that the process of the said application '2677 gives a dilute extract which is a ready-made formulation of the desired said constituents. Removal of the solvent from such a primary extract can directly give a mixture(or formulation) of the said constituents of interest either in the form of a concentrated solution or as a solid, or semi-solid residue. Solid form extract can also be obtained by adopting a suitable excipient/binder combination. Such extracts are suitable for direct administration to subjects. All such process variants yielding products in different forms such as a dilute extract, a concentrated extract, a solid form extract, a semi-solid extract, a treated extract and others are within the scope of the invention.

Thus, the product of the process of the invention may be a said dilute extract, a concentrated extract, a said solid or semi-solid residue or an extract that has been adsorbed on an excipient. The scope of the term 'dilute extract' also covers all the different extracts and extract streams that arise in a multi-stage extraction operation. TC extracts such as the ones described above may be subsequently treated with any of several physical and chemical operations/steps and also product finishing operations within the scope of the invention. The products arising from such treatments are all considered to be products of the process of the invention.

Clearly, this approach of the process of the invention offers the possibility of considerable savings in capital and operating costs, space requirements and also other advantages in the preparation of formulations. The principal advantage is the reduction in the number of processing steps required. All the advantages cited in 2677 also apply to the present invention. AH the process variants explored and that are possible therein have corresponding variants in the processes of the invention forming part of the current application. Such variants are within the scope of the invention. Said application 2677 provides for treatment of the plant matter to be extracted with a second plant matter(s) containing acidic components such as, for example, amla fruits. The botanical name of the amla plant is Emblica officinalis. This is also the highlight of the present invention wherein also the TC plant matter constituting said first plant matter is reacted with an acidic component containing second plant matters). The acidic plant matter adopted in the preferred embodiment herein is amla fhxit matter. It will be appreciated that within the scope of the invention, said second plant matter(s) may comprise any species having said acidic components), or mixtures of such species. The TC may be treated with the different said second plant matter species separately or together within the scope of the invention. As mentioned, said treatment may be carried out before the said extraction operation or simultaneously with the said extraction operation within the scope of the invention.

The choice of available acidic-component containing plant matter is large and, thus, this invention provides a large number of processing options to the process technologist. Thus, by a suitable choice of the acidic component containing plant species constituting said second plant matter(s) he can selectively get the desired TC principles into the extract. He can also aim to get the said selected TC principles in the extract in substantially the targeted/desired proportions. The invention provides the option to get the desired said constituents in the right form and in the right combination into the extract to give a substantially complete formulation. This invention provides for one or more additional steps to be optionally carried out on the extracts such as, for example, incorporation of additional compounds in the extract product, topping of the constituents to achieve the desired formulation composition, treatment to enhance biological availability and metabolic activity of the constituents, addition of components or other treatment to transform the extracts into dosage forms for application by different routes, or into any of the known pharmaceutical salts or adopt specific pharmaceutical carriers, conversion to obtain desired appearance, shape, colour, bulk, taste, flavour, odour, texture or other properties, addition of other therapeutical agents, treatment so as to obtain any pharmaceutical, nutraceutical, dietary or nutritional compositions/formulations/supplements incorporating the TC extract of the invention, carrying out of product finishing operations or others within the scope of the invention. These features and advantages of the application 2677 are also present in the present invention.

Said acidic components in the second plant matter(s) convert one or more of the said constituents that are insoluble in aq. ethanols into aq. ethanol-soluble forms which thus become extractable by the aq. ethanols adopted as solvents in the present invention. The same applies for the water solvent adopted in this invention. The wide choice of the different acidic components available in the form of said second plant matter(s) gives one the option to get the said insoluble components) in chemical form(s) which have other desired properties such as, for example, a greater degree of metabolic and biological activity and efficacy than the original form(s). It also provides the option to optimise the extraction/formulation process with reference to yield, processing time and other factors and achieve cost advantages. It also offers the option to obtain specific proportions of the constituents in the extract at the end of the extraction operation without requiring the conventional operation of the admixing of previously isolated constituents.

This invention provides for a process of extraction of TC plant matter by water, wherein the first plant matter, namely, T. connaroides plant matter is treated with a said second plant matter(s) that comprises acidic components. Said treatment is carried out either before, or during the extraction operation by the water solvent. This option also applies to the alcoholic solvent based process of the invention.

This invention also provides for a similar process of extraction of TC plant matter by a lower aliphatic alcohol such as ethanol or aq. ethanol. The preferred said second plant matter for the treatment of TC plant matter is amla fruit matter. However, the choice of the acidic plant matter(s) depends on the desired TC costituent(s) that require to be extracted. Within the scope of the invention, the solvent may comprise any mixture of the named solvents.

Alcohols, are very convenient and widely used solvents for plant extractions. Ethanol is particularly versatile and widely used solvent. These remarks apply to both pure alcohols and to their aqueous solutions and also to mixtures of alcohols and to the aqueous solutions thereof. Aqueous ethanol, as also non-aq. ethanol, is a particularly convenient solvent that also happens to be a good pharmaceutically acceptable carrier. Ethanol is generally edible and non-toxic, easy to handle and easy to remove from solutions where required. They are better and faster solvents for plant nutrients and dissolve a wider range of said nutrient solutes.

The adoption of aqueous alcohols as said single primary solvents, coupled with said acid treatment of T. connaroides by the present invention is novel and offers numerous technical advantages and cost benefits. It combines the advantages associated with both water and ethanol solvents.

These processes of the invention.offer a number of unexpected technical and techno-economic advantages.

The matter in this specification is not strictly and rigidly divided into the sections mentioned. For example, statements relating to the description and scope of the invention will be found distributed throughout the text. They will be found in sections other than the sections titled, 'Brief Description of the Invention' and 'Detailed Description of the Invention'. Similarly, statements relating to definitions of terms are also spread into different sections.

Definitions of Terms

In this specification, the scope of the term 'alcoholic solvent' includes, as mentioned,

(a) a single alcohol;

(b) a mixture of two or more alcohols;

(c) a solution of (a) in water or other suitable solvent; and

(d) a solution of (b) in water or other suitable solvent, wherein said alcohol(s) may be any of the aliphatic alcohols which may be liquid or solid at ambient temperatures.

Γη this specification, the acidic matter(s) selected for reacting with said nutrients may be in any of the following forms or combinations thereof:

(i) inorganic acids and/or inorganic acidic compounds,

(ii) organic acids and/or organic acidic compounds,

(iii) one or more plant matters containing organic acids and/or organic acidic compounds.

The term 'spent plant matter' is used both for partially and fully spent matter, herein. The extracted plant matter arising at any stage in a multi-stage extraction process is referred to herein as 'spent plant matter' or 'spent matter'. The meaning appropriate to the context may be taken.

The term 'extraction' is used herein to refer to both the overall process of extraction such as of plant matter comprising a series of steps including the actual extraction step(unit operation) wherein the plant matter to be extracted is contacted with the solvent or solvent mixture. Said extraction step may comprise leaching, soxhlet extraction or other known means of carrying out the extraction operation. Such extractions may be carried out at room temperatures or at elevated temperatures. An extraction operation may comprise a single extraction stage or a plurality thereof within the scope of the invention. The meaning appropriate to the context and offering the widest scope may be taken.

It is intended herein that the scope of the term 'solvent' also covers the adoption of a solvent mixture unless otherwise specified. The scope of the term also covers solutions of the solvents, or of solvent mixtures, in suitable solvents such as, for example, water. Thus, all aqueous alcohols are included in the scope of the term 'solvent' in so far as this invention is concerned.

The term 'plant nutrient' and 'nutrient' is to be understood broadly in so far as this specification is concerned to cover all phytochemicals constituting the plant matter(s) being extracted. The scope of the term includes all phytochemicals contained in the plant matter being extracted and particularly extends to all constituents in the plant matter relevant to food, dietary, nutrition, medicinal and therapeutical considerations. Specifically, it includes vitamins and minerals, antioxidants, carbohydrates, proteins and fats, medicinal principles and other compounds. It also extends to factors in the plant matter such as for odour, taste, colour and others. The term 'nutrients' also covers the precursors thereof, if any, present in the two plant matters.

In this specification, the terms 'acidic components', acidic reagents' and 'acidic compounds' are intended to be interchangeable and are intended to refer to either a free acid(s) or an acidic substance(s) or both. Said acidic reagents may be inorganic or organic. Use of mixtures of organic and inorganic origin acidic components is also within the scope of the invention.

Preferably, the acidic components are in the form of plant matter. The term 'reagent' is intended to mean a reactive compound herein.

Brief Description of the Invention

According to the invention, therefore, there is provided a method/process of extraction of medicinal principles, vitamins and minerals and other phytochemical constituents from Trichilia connaroides, wherein a first plant matter comprising one or more parts of Trichilia connaroides plant is extracted with a solvent, or solvent mixture, to yield an extract solution, and characterised in that said first plant matter is treated with a second plant matter(s) containing acid(s)/acidic compound(s) such as to convert one or more of the solvent-insoluble constituents of the former matter into form(s) that are soluble in said solvent and/or to convert one or more of the soluble constituents therein into forms having increased solubility.

Detailed Description of the Invention

Within the scope of the invention, a said extraction operation may comprise a single stage or a plurality thereof. Different flow arrangements of the various process streams such as of the incoming plant matters, the solvent, the partly and fully spent plant matters and the different streams of the extract solutions of different concentrations are feasible in multi-stage extraction operations and the same are within the scope of the invention. The extract generated at any point in such a multi-stage extraction system is a rpoduct of the process of the invention.

Thus, this invention provides for the adoption of alcoholic solvents. The term 'alcoholic solvent' is understood as far as this specification is concerned, to be an alcohol or any solution thereof. The alcohol may be any of the aliphatic alcohols or mixtures thereof. It is further intended to include within the scope thereof, any solution of an alcohol, or of an alcohol mixture in a suitable solvent such as for example, water. Thus, aqueous alcohols of all strengths are included in the scope of the term 'alcoholic solvents' of this invention. The alcohol(s) in an alcohol solution of this invention may be one that is a liquid at ambient temperatures or that is solid. Aqueous ethanol is the preferred solvent of the present invention.

The adoption of aqueous alcohols as said single primary solvents, and particularly aqueous ethanol coupled with said acid treatment of the plant matters by the present invention is novel and offers numerous technical advantages and cost benefits. It combines the advantages associated with both water and ethanol solvents.

The reaction of conversion by said acidic component(s) may be carried out before or during the extraction operation on the first plant matter. In one embodiment, the said first and second plant matter(s) are pulped and mtermixed such that the said reactions occur in the solid phase. The partly or fully reacted pulp mixture is then contacted with the extraction solvent. A liquid, such as the adopted solvent, may be incorporated in the pulped mixture for better contact between the two plant matters.

In another embodiment, the reactions are carried out in a liquid phase. The two plant matters are dispersed in powder form in a liquid. Preferably, the adopted extraction solvent forms the said liquid phase wherein the two plant matters, first and second, are dispersed. A number of contacting and cooking methods are known in the art and may be adopted for carrying out said reaction(s) and or the extraction operation.

In this invention, the TC plant matter constitutes said first plant matter, a set of the medicinal and other factors of which require to be extracted as comprehensively as possible and to the maximum extent possible. Towards this objective, this invention provides a second plant matter(s) that comprises suitable acidic components that react with the first plant matter to convert as many as possible of the solvent-insoluble factors therein into soluble forms. The object of this invention is also to select such said second plant matter(s) that it preferably renders one or more of the solvent-soluble principles therein into forms that are more soluble. It is also the object of the invention that the said acidic components are such that preferably the converted forms of the said solvent-insoluble principles are biologically and metabolically active and if not, are convertible into such forms by an additional treatment of the treated first plant matter. The solvent referred to herein is the adopted solvent. The roles of said first and second plant matter(s) are not intended to be mutually exclusive. Each may constitute a source of said medicinal principles to be extracted and of the said acidic reactants. The combination of the two plant matters, said first and second, is preferably such as to give an extract that contains as many of the medicinal factors required, said factors coming from either the said first plant matter or the second or both. The extracted nutrient(s) may come from said second plant matter(s) in addition to said first plant matter within the scope of the invention. Also, said acidic component(s) present, if any, in said first plant matter may take part in the said conversion reaction(s) involving the said nutrient(s) present in the first plant matter, or me second, or both, within the scope of the invention.

Preferably, the adoption of the solvent coupled with said acidic treatment should be such as to give increased overall extraction yield and the yields of the specific medicinal and other factors being extracted. Preferably, the selection of said second plant matter and the acidic factors thereof is such as to make the extraction process simpler and quicker and yield components in forms that are more biologically and metabolically active and efficacious.

One object of the invention is to obtain an extract that comprises the desired combination of the different plant constituents in the desired proportions, or as close thereto as possible, such that subsequent treatment and processing steps to obtain the final formulation are reduced/minimised.

Within the scope of the invention, said acidic treatment of the first plant matter may comprise addition of free acids thereto or a combination of said second plant matter(s) together with free reagent acids or other acidic compounds may also be adopted within the scope of the invention. Said free acids may be inorganic or organic. Preferably, the acidic matter is in the form of said second plant matter(s) comprising plant-based organic acids and organic acidic compounds. The term 'reagent' is meant to refer to a reacting compound in the free form rather than in a combined form in plant matter(s). Within the scope of the invention, said first plant matter may comprise other plant species in addition to T. connaroides. Said second plant matter may also comprise a mixture of species within the scope of the invention. The two plant matters, said first and second, may each comprise a single plant part such as, for example, fruits, roots, leaves, seeds, bark, flowers or others or mixtures of plant parts. Thus, this invention provides for a water extraction of T. connaroides plant matter combined with adoption of a suitable said second plant matter(s), which gives a better yield of the TC principles than by the use of water alone. Similarly, this invention provides for aqueous ethanol extraction of TC plant matter together with a suitable said second plant matter(s), which gives a higher yield of the TC principles than by the use of aq. ethanol solvent alone. The process of the invention, can be used also with the other lower alcohols such as methanol, propanols and butanols. Non-aq alcohols also may be used within the scope of the invention. The said yield is better with the ethanol-amla process over the water-amla process of the invention.

In order to provide a better understanding of the invention and without limitation to the scope thereof, two embodiments thereof are now described in detail hereinbelow.

The process of the invention adopted for the extraction of T. connaroides with water using amla fruits as second plant matter comprises the following steps.

The process involves the extraction of Trichilia connaroides aerial parts along with a synergistic quantity of Amla fruits with water to yield an extract.

Powder ( Hammer mill)

Φ

Mix about 5 kgs of Amla fruits crushed, with about 100 legs of Trichilia dried aerial

parts

Extract with WATER exhaustively at about 40-98°C for about

6 hours by boiling, for about three times (Stainless Steel Extractor) - ψ

Concentrate I extract to about 1/10* volume, this is A (Concentrator / thin film evaporator) Φ

Concentrate II extract to about 1/10^Λ volume, this is B (Concentrator / thin film evaporator) Φ

Preserve the IE extract for extraction of next batch (Stainless Steel tank)

Ψ

Combine A and B (Reactor with agitator)

Ψ Filter over a filter bed (Leaf filter / Filter press / Sparkler /Centrifuge)

Ψ

Concentrate the filtrate to about l/5^tb volume (Concentrator / thin film evaporator)

Add an excipient to adsorb the liquid, and mix well (Planetary mixer / Rapid Mixer Granulator/ Slurry Tank)

Ψ

Spread it on trays and dry(Tray drier / Vacuum Drier/ Fluid Bed Drier )

Alternately spray dry it (Spray Drier)

Powder (Stainless Steel Multi mill / Hammer Mill / Pulveriser)

Sieve (Stainless Steel Sifter / Sieve Shaker)

Ψ

Blend entire batch quantity (Stainless Steel Double Cone Blender / Ribbon Blender / Octagonal Blender)

Trichilia connaroides - Water extract (TWSE)

For comparison, TC plant matter was extracted following a similar procedure as given above but without the adoption of the second plant matter containing acidic components, namely, amla fruits. The overall yield of the TC principles following the adoption of the acidic second plant matter, amla fruits, that is for the water-amla system was observed to be about 12% by wt. The yield of the saponins and terpenes was about 12.3% by wt and about 3.9% by wt. The corresponding figures for water extraction without the adoption of the second plant matter(amla fruits matter) were: about 10%, about 8.2% and about 1.1% by wt. respectively.

The process of the invention adopted for the extraction of T. connaroides with aq. ethanol using amla fruits as second plant matter comprises the following steps.

The process involves extraction of Trichilia connaroides aerial parts along with synergistic quantity of Amla fruits with about 50% aq(aqueous) ethanol to yield an extract.

Φ

Powder(Hammer mill)

Φ ■

Mix about 5 kgs of Amla fruits crushed, with about 100 kgs of Trichilia dried aerial parts Extract with about 50% Ethyl alcohol exhaustively at about 40-80°C for about

6 hours by boiling, about three times (Stainless Steel Extractor)

Φ

Concentrate I extract to about 1/10^th volume, this is A (Concentrator / thin film evaporator) Φ

Concentrate Π extract to about 1/10^th volume, this is B (Concentrator / thin film evaporator) Φ

Preserve the III extract for extraction of next batch (Stainless Steel tank)

Φ

Combine A and B (Reactor with agitator)

Φ

Filter over a filter bed (Leaf filter / Filter press / Sparkler /Centrifuge)

Φ

Concentrate the filtrate to about 1/5^Λ volume (Concentrator / thin film evaporator)

Φ

Add an excipient to adsorb the liquid and mix well (Planetary mixer / Rapid Mixer Granulator/ Slurry Tank)

Φ

Spread it on trays and dry (Tray drier / Vacuum Drier/ Fluid Bed Drier )

Alternatively spray dry the material (Spray Drier)

Φ

Powder (Stainless Steel Multi mill / Hammer Mill /Pulveriser )

Φ

Sieve (Stainless Steel Sifter / Sieve Shaker)

φ

Φ

Trichilia connaroides - Alcohol Synergy extract (TASE)

The overall yield of the TC principles in the aq. ethanol extract following the adoption of the acidic second plant matter, amla fruits was 14% by wt. The yield of the saponins and terpenes was 15.6% by wt and 5.5% by wt. The respective figures for aq. ethanol extraction without the adoption of the amla second plant matter were: 8%, 11.1% and 4.1% by wt. respectively. For comparison, TC plant matter was extracted following the same procedure as given above but without the adoption of the second plant matter containing acidic components, namely, amla fruits. It will be observed that the overall and individual constituent yields are better for the process of the invention employing amla second plant matter than the conventional process which does not involve the use of said acidic plant matters. This is true for both water extraction and extraction by 50% v/v aq. ethanol. Between the process of the invention involving water solvent and amla fruit matter and involving 50% v/v aq. ethanol solvent and amla fruit matter, the yields are better in the latter.

The yield of saponins and terpenes in the extracts of the invention compared to said counterpart extracts is greater. Some of the steps in the abovementioned process flowsheets are optional. Thus, other flowsheets are possible and are within the scope of the invention. For example, the no. of stages and the material flow arrangements/patterns shown are also optional and many other

arrangements are possible. All such variants feasible in the art are all within the scope of the invention.

Suggestions of the equipment to be adopted for the various operations are also optional and any suitable equipment available from the wide range in the art may be adopted. Such variants are also within the scope of the invention. Embodiments and variations other than described herein above are feasible by persons skilled in the art and the same are within the scope and spirit of this invention.

References

1. Puroshothaman KK, et al ( 1983) Structure of Heynic Acid - A new triterpene acid from Heynea Trijuga Roxb. Ind. J of Chem, 1983:22B: 820-1.

2. Puroshothaman KK, et al (1987) Trijugins A and B, Tetranor triterpenoids with a novel rearranged carbon skeleton from Heynea trijuga Roxb. Can. J of Chem. 1987:65:35-7.

3. Mathuram V, Kundu AB, (1990) Isolation and Characterisation of Trijugin B acetate from Heynea trijuga Roxb. Indian J of Chem, 1990:29B:970.

4. Zhang SP, et al, (2003) A pentanortriterpenoid with a novel carbon skeleton and a new pregnene from Trichilia connaroides, Can. J of Chem, 2003:81(3):253-7.

5. Prasanna GS, Purnima A, (201 lB),Antihyperlipidemic effect of Trichilia

connaroides in hypercholesterolemic rats and its possible mechanism, J of

Pharm. & Bioall. Sci. (Apr-Jun 201 l):Vol 3: Issue 2:230-235.

6. Inada et al, (1994) Structures of a new limonoid and new triterpenoid derivative from pericarps of Trichilia connaroides, J Nat Prods, 57, 1446-1449.

7. Wang et al, (2008) Structural elucidation of limonoids and steroids from Trichilia

connaroides, Phytochem, 69, 1319-1327.

Geng ZL et al, (2009) Trichilin B, a novel limonoid with a highly rearranged ring system from Trichilia connaroides, Tetrahedron Letters, 50, 2132-2134.

9. Agarwal G et al,(2010), Volatile constituents of Trichilia connaroides( Wight and

Am.) roots, Asian J Trad. Medicines, 2010:5(5).

10. Purnima AL, Mathuram V, (2003A) Indian J of Pharm. Sci, 2003, 65, 37.

11. Purnima A, Prasanna GS, Mathuram V, (2006) Analgesic and anti-inflammatory activity of the chloroform extract of Trichilia connaroides(W & A) Bentilizen, Indian J. Pharm. Sci, 2006:68:231-233.

12. Prasanna GS, Purnima A, (2011 A) Antikhyperhomocysteinemic and

antihyperlipidemic effect of Trichilia connaroides in methionine-induced

hyperhomocysteinemic animals, Ind. J of Pharmacol. 2011 April, 43(2):203-206.

13. Tandon S, et al (2009) Growth-regulatory activity of Trichilia connaroides(syn.

Heynea trijuga) leaf extracts against the Bihar hairy caterpillar Spilosoma

obliquia(Lepidoptera: Arctiidae), Intern. J of Tropical Insect Sci, Vol.29, No.

4,pp 180-184(2009).

14. Purnima AL, Mathuram V, (2003B) Hypocholesterolemic and

antihypercholesterolemic activity of extracts of Trichilia connaroids on rats. Ind.

J of Pharm. Sci. Sep-Oct 2003, 537-539.

Claims

Claims:

1. A method/process of extraction of medicinal principles, vitamins and minerals and other phytochemical constituents from Tnchilia connaroides, wherein a first plant matter comprising one or more parts of Tnchilia connaroides plant is extracted with a solvent, or solvent mixture, to yield an extract solution, and characterised in that said first plant matter is treated with a second plant matter(s) containing acid(s)/acidic compound(s) such as to convert one or more of the solvent-insoluble constituents of the former matter into form(s) that are soluble in said solvent and/or to convert one or more of the soluble constituents therein into forms having increased solubility.

2. The method/process of extraction of medicinal principles, vitamins and minerals and other phytochemical constituents from Trichilia connaroides, as claimed in the preceding claim 1, and wherein said first plant matter and or the said second plant matters) are subjected to one or more preparatory treatments such as, for example, selection, picking, cleaning, washing, blanching, cutting, dicing, pulping, liquidising, drying, crushing, grinding, puowdering/pulverising and others.

3. The method/process of extraction of medicinal principles, vitamins and minerals and other phytochemical constituents from Trichilia connaroides, as claimed in any of the preceding claims 1 and 2, and further comprising carrying out of one or more of the undermentioned operations on the extract towards the conversion and/or finishing of the extract to yield a pharmaceutical, food or nutraceutical composition, formulation or supplement,

(i) division into desired dosages;

(ii) conversion into any of the dosage forms required for administration to subjects by different routes;

(iii) conversion such as to impart biological and metabolic availability, activity and

efficacy or to

enhance the same;

(iv) conversion of the said nutrient(s) into any of the known pharmaceutical salts;

(v) adaptation of the extract to any of the known pharmaceutical carriers;

(vi) conversion of the extract by addition of required components to obtain a specific form, appearance, consistency, particle size, colour taste, flavour texture and other properties in the product;

- 1 - (vii) incorporation of other nutrient(s) and/or topping of nutrient(s) to bring them up to the desired levels or such as to conform to medicinal, food, dietary or nutraceutical standards, such as for example, the RDA(Recommended Daily Allowance) requirements;

(viii) incorporation of other food, medicinal and nutrition factors towards obtaining the desired composition, formulation or supplement;

(ix) incorporation of therapeuticaly active components); and

(x) one or more physical and/or chemical operation for the finishing of the product,

(xi) and others.

4. The method/process of extraction of medicinal principles, vitamins and minerals and other phytochemical constituents from Trichilia connaroides, as claimed in any of the preceding claims 1 to 3 wherein said treatment of the said first plant matter by acid(s)/acidic(compound(s) is carried out substantially during the said extraction thereof by said solvent solvent mixture.

5. The method/process of extraction of medicinal principles, vitamins and minerals and other phytochemical constituents from Trichilia connaroides, as claimed in any of the preceding claims 1 to 4, wherein said second plant matter(s) comprises amla fruit matter.

6. The method/process of extraction of medicinal principles, vitamins and minerals and other phytochemical constituents from Trichilia connaroides, as claimed in the preceding claim 5, wherein the said solvent comprises water.

7. The method process of extraction of medicinal principles, vitamins and minerals and other phytochemical constituents from Trichilia connaroides, as claimed in the preceding claim 6, wherein the ratio of the amounts of said amla fruit matter to the Trichilia connaroides plant matter is from about 3 kg. to about 7 kg. of the former to about 100 kg. of the latter.

8 The method/process of extraction of medicinal principles, vitamins and minerals and other phytochemical constituents from Trichilia connaroides, as claimed in the preceding claim 7, wherein the said ratio is about 5 kg. of the said amla fruit matter to about 100 kg. of the said Trichilia connaroides plant matter.

9. The method/process of extraction of medicinal principles, vitamins and minerals and other phytochemical constituents from Trichilia connaroides, as claimed in any of the preceding claims 6 to 8, wherein the extraction temperature is maintained from about 40 C to about 98 C.

10. The method/process of extraction of medicinal principles, vitamins and minerals and other phytochemical constituents from Trichilia connaroides, as claimed in the preceding claim 5, wherein the said solvent/aolvent mixture comprises aqueous ethanol.

11. The method/process of extraction of medicinal principles, vitamins and minerals and other phytochemical constituents from Trichilia connaroides, as claimed in the preceding claim 10, wherein the said aqueous ethanol solvent is of strength from about 10% v/v to about 95% v/v.

12. The method/process of extraction of medicinal principles, vitamins and minerals and other phytochemical constituents from Trichilia connaroides, as claimed in the preceding claim 11, wherein the strength of the aqueous ethanol solvent adopted is about 50% v/v.

13. The method process of extraction of medicinal principles, vitamins and minerals and other phytochemical constituents from Trichilia connaroides, as claimed in any of the preceding claims 10 to 12, wherein the extraction temperature is maintained from about 40 C to about 80 C.

14. The method/process of extraction of medicinal principles, vitamins and minerals and other phytochemical constituents from Trichilia connaroides, as claimed in any of the preceding claims 10 to 13, wherein the ratio of the amounts of said amla fruit matter to the Trichilia connaroides plant matter is from about 3 kg. to about 7 kg. of the former to about 100 kg. of the latter.

15. The method process of extraction of medicinal principles, vitamins and minerals and other phytochemical constituents from Trichilia connaroides, as claimed in the preceding claim 14, wherein said ratio is about 5 kg. of the said amla fruit matter to about 100 kg. of the said Trichilia connaroides plant matter.

16. The methods/processes of extraction of medicinal principles, vitamins and minerals and other phytochemical constituents from Trichilia connaroides, substantially as hereindescribed.

17, An extract made by the method/process of extraction of medicinal principles, vitamins and minerals and other phytochemical constituents from Trichilia connaroides, as claimed in any of the preceding claims 1 to 16.

18. A pharmaceutical, food, dietary or nutraceutical composition, formulation or supplement, incorporating an extract as claimed in CI. 16, or an extract made by the method/process of extraction of medicinal principles, vitamins and minerals and other phytochemical constituents from Trichilia connaroides, as claimed in any of the preceding claims 1 to 16.