FORM 2
THE PATENT ACT, 1970 (39 OF 1970)
COMPLETE SPECIFICATION (See Section 10)


" Powder of the Roots of Ampelocissus
latifolia Roxb Plant used for treatment
of gynecological problems "
Bharati M. Rao; an Indian of A/101, Alka Apartment, Opp. M.A. High School, S.V. Road, Andheri (West),
Mumbai- 400058
The following specification particularly describes the nature of the invention and the manner in which it is to
be performed.


This refers to a chemical compound obtained from Ampelocissus latifolia Roxb., plant belonging to family Vitaceae, which is also known as Vitis latifolia Roxb. This plant grows in farms in South Gujrat. It is not Perennial Plant available in all the seasons. It is seasonal plant and is available in the monsoon season after one month rain. It is grows and is available upto two or three months in the monsoons season only. When the rains stop these plants of Ampelocissus latifolia Roxb. dry up and after few months they are not visible. With the onset of the monsoon plants come out from the same roots again. If the roots are dug out the plant will not grow again.
Ampelocissus latifolia Roxb., plant belonging to family Vitaceae is also known as Vitis latifolia Roxb. It is a climber with annual stems on 3-6 m long, green, often tinged purple especially at the nodes, covered with a thin glaucous bloom. The Plant is found in subhimalayan tract upto Maharashtra and Western Ghats. The berried though not eaten by humans are known as wild grapes and are edible (Kirtikar & Basu, 1975; Anonymous, 1972).
Ute plant has been reported to possess medicinal activities. Hae raots of the plant species are astringent and taken to cure dfammc dysentery (Anonymous, 1967) and also used for curing ifractaies., myalgia, pneumonia, snakebite and sore (Johnson, JL9http: / / www, holisticopia. com / herbage / files / H86. htm# 12 72L TEhe whole plant is also used an alternative, astringent aed for odontalgia and sore (http: / /www.ars-grin.gov/cgibin/diike.ethnobot.pll.
Hfoera asrc 3aaany ethnomedicinal practices that make use of the spentes.. Moot extract (from one inch root-piece) is given to (drank fer poisonous snake bite (cobra or krait), for curing toners, applied locally on boils, skin lesions and back pain. M©©£ pieces are tied to horn of cattle for skin lesions, infections anas! tfifae paste is applied to cure veterinary fractures, (dM©ca±bns, broken horns (D' Cruz, 2002).
The present patent is for the roots of the plant Ampelocissus latifolia Roxb being used to cure gynecological disorders. It is
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given in form of a paste with honey to women who fail to conceive or face abrupt miscarriages in their pregnancy and excessive bleeding during menopause.
Another species belonging to the same genus, Vitis vinifera contains reservatol, a potent phytoestrogen that displays similar gynecological activities (Lamuela-Ravento, 1995).
Phytoestrogens are now being increasingly used as an alternate method of medication for sexual disorders, mainly because the hormone replacement therapy, which was thought to be highly beneficial, has not shown enough promising result. (Hays et al., 2003).
Micropropagation studies repot plantlet regeneration via somatic embryogenesis using anther culture technique (Salunke, 1999). However, to the best of our knowledge, no direct organogenesis in the species has been reported.
No Phytochemical analysis was found to be reported in the existing literature.
The roots of the plant are picked from the farms and then are cut into small pieces and dried in the sunshine. After the roots dry up they are grounded to form a fine powder.
The powdered root of the plant of Ampelocissus latifolia Roxb is used for treating menstrual dismenorrhea, infertility among women.
The women who are unable to conceive naturally, on taking this powder of the roots of Ampelocissus latifolia Roxb with half teaspoonful sugar and ghee are able to conceive. The powder has to be taken early in the morning on empty stomach as herbal medicine and within short period they could conceive. Those women who were facing miscarriage problem were also taking this powder during their pregnancy and could solve their problem and had a healthy baby after nine months.
During Manopause, women can take the powder of the root of Ampelocissus latifolia Roxb to stop excess continuous bleeding
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immediately. This powder of the roots of Ampelocissus latifolia Roxb is known to have miracle effect in women who are troubled by discomfort and excessive bleeding during menopause.
Women who were administered this powder, on second day only the excessive bleeding stopped without any other medicine. This powder of the roots of Ampelocissus latifolia Roxb is helpul in a miracle solution in reducing the women's discomfort and embarrassment experienced over the years due to excessive bleeding, weakness and mood swings in menopause. It is a good and cheaper alternative to the invasive and expensive hystectomy procedure advised by Doctors.
Its yet another benefit is experienced by those ladies who have taken this root powder during pregnancy they feel energetic. This powder gives them strength and it can be used as a Tonic. This powder can remove all types of problems like Jaundice, Skin infections of the children in the womb.
This root contains the steroids in measurable amounts. Root is used in case of infertility as well as in menopause bleeding problem and root has the properties to help in the above gynecological problems. Women prefer to have this plant based steroids which is a natural ingredient and has no side effects unlike other allopathic medicines.
The Phytochemical and plant tissue culture chemicals studies and large scale cultivation of this plant are as under: -
Materials and Methods :
General Methodology:
The study was mainly divided in three steps :
• To carry out qualitative Phytochemical analysis of dried roots f the plant Ampelocissuse latifolio Roxb.
• To track down components of biological significance by Thin Layer Chromatography,
• To establish in vitro callus cultures to further verify presence of similar compounds in the callus.
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Qualitative Phytochemical analysis of dried root power of Ampelocissus latifolia Roxb.
The root powder was stored in an airtight container away from sunlight to protect its content from getting affected.
To establish the presence of various group of Phytochemical, qualitative tests were carried out using standard methods.
Alkaliods: Presence of alkaloids was checked by testing acidified residues obtained by treating dried powder with ammonical ethanol with reagents like Mayer's reagent, Dragendroffs reagent and Wagner's reagent. Precipitation indicated presence of alkaloids.
Tannins and Saponins: Aqueous extract of the dried plant materials was used to check the presence of tannins and saponins, by precipitating with gelatin and shaking vigorously to produce froth respectively.
Flavonoids: Colour change of the acidic extract, on treating with HCI and Mg ribbon, was observed for the presence of flavonoids.
Iridoids: Presence of iridoids was checked by the presence of red colour of the acidified extract with Trim-Hill reagent. Appearance of red colour in the layer containing Isoamyl alcohol on mixing the acidic extract with the same indicate presence of iridoids.
Steroidal compounds(steroids,Triterpenoids, terpenoids etc.):
Liebermann Burchard test was carried out to check the presence of triterpenoids and steroidal compounds. Production of purple colour with acetic anhydride and Sulphuric acid indicated the test to be positive for die steroidal compounds.
Thin Layer Chromatography to track down components of biological significance:
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Extraction of root powder was done in methanol before and after hydrolysis. Hydrolysis was carried out by refluxing powder in 2 N HCI for around 5-6 hrs. followed by neutralization using Sodium Carbonate. This powder, after drying was extracted by refluxing in methanol.
TLC plates were spread and activated at 110c. and the sample was applied. The plates were then run in the solvent systems (). It was then sprayed with Sulphuric acid and purple coloured bands counted and Rf was taken for both methanolic as well as hydrolyzed extracts.
In vitro callus cultures of Ampelocissus latifolia Roxb . :
To establish callus cultures, both leaves as well as nodes were used as explants and were inoculated on MS medium fortified with different phytohormones like NAA and BAP. BAP were taken in combination with NAA and was also checked for its effect individually.
Results :
Qualitative Phytochemical analysis of dried root powder of Ampelocissus latifolia Roxb*
Preliminary Phytochemical Analysis of the dried root powder of Ampelocissus latifolia Roxb. Was carried out in triplicates and the result were recorded upto the scale of +++.
The result indicated higher concentrations f Tannins, Saponins, Flavonoids, Proanthocyanidines and Iridoids, while comparatively lower concentrations of Alkaloids were observed int eh dried sample of root powder of Ampelocissus latifolia Roxb. In its qualitative analysis (Table. 1)
Twin Layer Chromatography to track down component of biological significance :
On the basis of available literature on Thin Layer Chromatography (Daniel, 1985; Evans,), various extracts were prepared. These extracts were tested for the presence of
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steroidal compounds using Thin Layer Chromatography with various solvent systems out of which Choloroform : Ethyl Acetate (1 :1), Hexane: Acetone (4:1) showed the best separation. Spraying reagent used was 50% Sulphuric Acid and Liebermann-Burchard reagent.
1. Methanolic extract : obtained by refluxing the dried root powder in Methanol for around an hour.
2. Chloroform extract : obtained by refluxing the hydrolyzed root powder.
3. Petroleum Ether extract : obtained by refluxing the hydrolyzed root powder.
Table . 2 highlights the response that these extracts showed with the solvent system ().
In vitro callus cultures of Ampelocissus latifolia Roxb.:
To establish callus cultures, both leaves as well as nodes were used as explants and were inoculated on MS medium fortified with different phytohormones like NAA, 2,4-d and BAP. BAP were taken in combination with NAA and was also checked for its individually.
Tissue culture requires fresh plant materials and the plant species, Ampelocissus latifolia, being a seasonal one was available only during the monsoon season. This posed a major problem of contamination of fungi. Various surface sterilants were used and the right duration and concentration of surface sterilants were standardized.
The best axenic cultures were obtained using 1% Teepol (Soap solution) for 30-40 minutes, 0.5% Bavistin (Carbadezim) for 20 minutes and 0.1% HgC12 for 7 minutes followed by a minute long dip in 50% Alcohol. Each treatment was followed by repeated washes with autoclaved distilled water.
The powder of the root of Ampelocissus latifolia Roxb is a cheaper and beneficial option without side effects for treatment of gynecological problems among women.
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I Claim:
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1. The chemical compound found in the root of the Ampelocissus latifolia Roxb plant which when administered in the powdered form with honey and ghee is able to reduce the excessive bleeding during menopause;
2. The chemical compound as claimed in claim 1. used for treatment of miscarriages in pregnant women;
3. The chemical compound as claimed in claim 1. is a economical and better option then invasive hystectomy.