Title of Invention

"A WATER BASED SPERMICIDAL VAGINAL CONTRACEPTIVE COMPOSITION "

Abstract

The present invention relates to a method of preparing an aqueous composition useful as anti-spermicidal vaginal contraceptive, said composition comprising hydrogen peroxide of concentration ranging between 1 to 3%, and sodium chloride of concentration ranging between 0.4% to 3.0% with pH ranging between 4 to 4.5 and showing synergistic contraceptive effect, and said composition is an inexpensive, non-messy, fats acting vaginal contraceptive, with 100% efficacy but at the same time no side effects, and a method of producing like aqueous contraceptives.

Full Text

Field of the present invention The present invention relates to a method of preparing an inexpensive, non-messy, safe and fast acting aqueous spermicidal composition of 100% efficacy useful as a vaginal contraceptive free of side effects, said method comprising steps of preparing sodium chloride solution of concentration ranging between 0.4 to 3% in water, mixing hydrogen peroxide to said sodium chloride solution to obtain a mixture having hydrogen peroxide in the concentration ranging between 1 to 3%, adjusting pH of the mixture ranging between 4 to 4.5, and obtaining said composition, and also, the composition thereof. The said composition is capable of maintaining vaginal pH and has no adverse effect on the beneficial vaginal flora. Background and prior art references Maintenance of functional integrity of the spermatozoa is vital for successful fertilization with the oocyte. Impairment with any of the functional properties of the sperm might lead to affect the ultimate process of fertilization. Development of a contraceptive composition that affects majority of sperm functions is always considered more efficacious. Sperm motility is one of the important functions of the sperms, which enables the sperms to reach the oocytes for fertilization. Spermatozoa without forward progressive motility will not be able to perform this ultimate task, a key aspect while making a contraceptive composition. Reactive oxygen species (ROS) inhibit sperm movement and have been implicated in male fertility (Armstrong JS, Rajasekharan M, Chamulitrat W, Gatti P, Helistrom WJ and Sikka SC (1998) Characterization of reactive oxygen species induced effects on human spermatozoa movement and energy metabolism. Free Radical Biology Medicine 26: 869- 880) There are two sources of oxidative stress that affect human spermatozoa. The first source is the reactive oxygen species generated by the defective spermatozoa due to some defect in the spermatogenesis. The second source is from seminal leukocytes, infiltration of which is a common feature of the human ejaculate (Gavella M, Lipovac V, Vicic M and Rocic B (1996) Superoxide anion scavanging capacity of human seminal plasma. Ml J of Andrology 19: 82-90). The deleterious effects of ROS on sperm functions have been reviewed by Aitkin et al, 1994a. Aitkin RJ (1994) A free radical theory of male infertility. Reproduction Fertility and Development 6: 19-24. Hydrogen peroxide is the most common and toxic ROS to spermatozoa till recognized. Hydrogen peroxide induced cell death was reported in cerebellar granule cells, which was characterized by chromatin condensation and DNA fragmentation. During the process, the fluidity of the cell membrane decreased markedly and the confirmation of the membrane proteins altered significantly (Wei T, Ni Y, Hou J, Chen C, Zhao B and Xin W (2000) Hydrogen peroxide-induced oxidative damage and apopotosis in cerebellar granule cells: Protection by Ginkgo Biloba extract. Pharmacological Research 41: 427- 433.). Use of hydrogen peroxide has also been reported (patent no. 778 886 by Shihata et al, Shihata (1998) vaginal contraception compositions combining a spermicidal agent and a peroxygen compound. United States Patent No 778 886.) along with the known spermicidal Nanoxynol -9 to restore the alterations to pH caused by the spermicidal application affecting the vaginal flora itself. A letter to the editor {Strohmer et al in Human reproduction 1997 Jul; 12(7):1599}cites a case of a 37 year old patient who was using 3 % H2O2 soaked in cotton and inserting deep into vagina to the cervical os to avoid pregnancy. Applicants work is different from the work of Strohmer et al in many aspects and a few of such aspects are given below: 1. The contraceptive composition in instant application has normal saline along with H2O2 concentration ranging between 1 to 3%. Therefore the beneficial effects of the saline could neutralize the cyto-toxic effects of ^62 on the vaginal epithelium as observed in said letter to editor. 2. The efficacy of the method as stated by Strohmer et al. (1997) is questionable since putting a soaked cotton deep into the vagina could also block the opening of the cervical os preventing the sperms to pass through. Therefore, the resultant effect can be due to mechanical blockade and need not necessarily be due to H2O2 only. 3. No validation of the effect was submitted in the report by Strohmer (1997). 4. The contraceptive composition of instant application has synergistic spermicidal effect, with toxicity towards the sperms but having no irritation or any other adverse effects on vaginal epithelium and is safe for application on animal including humans. Concentrations of hydrogen peroxide 100 to 200 ul are reported Daru et al (effects of hydrogen peroxide on NDA and plasma membrane integrity of human spermatozoa; Fertility and sterility 74 (6): 1200-1206) to increase the oxidative stress on human sperms affecting their progressive motility and plasma membrane integrity and inducting DNA fragmentation. Also, there is a need for development of new and low cost contraceptives with greater efficacy, safety, and acceptability. Available vaginal contraceptives are oily preparations, either gel or foam based. These become messy during and following the coitus and as a result, the sexual pleasure is also claimed to be affected. This may be the reason for which the acceptability of such contraceptives is on decline. Therefore, developing a new, water-based contraceptive with good efficacy would go a long way in increasing the acceptability of the vaginal contraceptive use in the country. Objects of the present invention The main object of the present invention is to develop an effective vaginal contraceptive. Another object of the present invention is to develop a safe vaginal contraceptive. Yet another object of the present invention is to develop a vaginal contraceptive with an ability to bring down the sperm motility. Still another object of the present invention is to develop a non-messy vaginal contraceptive. Still another object of the present invention is to provide a shelf-stable composition. Further object of the present invention is to provide a method to develop a safe aqueous vaginal contraceptive with spermicidal activity. Another main object of the present invention is to develop a simple method of preparing an efficient vaginal contraceptive with spermicidal activity. In addition, another main object of the present invention is to develop a method of treating sperms with the synergistic spermicidal composition. Further, another main object of the present invention is to use the synergistic spermicidal composition of hydrogen peroxide and sodium chloride as vaginal contraceptive. Summary of the present invention The present invention relates to a method of preparing an inexpensive, non-messy, safe and fast acting aqueous spermicidal composition of 100% efficacy useful as a vaginal contraceptive free of side effects, said method comprising steps of preparing sodium chloride solution of concentration ranging between 0.4 to 3% in water, mixing hydrogen peroxide to said sodium chloride solution to obtain a mixture having hydrogen peroxide in the concentration ranging between 1 to 3%, adjusting pH of the mixture ranging between 4 to 4.5, and obtaining said composition, also, the composition thereof. The said composition is capable of maintaining vaginal pH and has no adverse effect on the beneficial vaginal flora. Statement of invention: Accordingly, the present invention relates to a water based spermicidal vaginal contraceptive composition comprising i. hydrogen peroxide of concentration ranging from 1 to 3%, and ii. sodium chloride of concentration ranging between 0.4 to 3%, with pH ranging between 4 to 4.5 showing synergistic contraceptive effect; and The present invention also relates to a method of treating sperms with an inexpensive, non-messy, safe and fast acting aqueous synergistic spermicidal composition of claim 16 of 100% efficacy as vaginal. Detailed description of the present invention Accordingly, the present invention relates to a method of preparing an inexpensive, non-messy, safe and fast acting aqueous synergistic spermicidal composition of 100% efficacy useful as a vaginal contraceptive free of side effects, said method comprising steps of preparing sodium chloride solution of concentration ranging between 0.4 to 3% in water, mixing hydrogen peroxide to said sodium chloride solution to obtain a mixture having hydrogen peroxide in the concentration ranging between 1 to 3%, adjusting pH of the mixture ranging between 4 to 4.5, and obtaining said composition, also, the composition thereof. The said composition is capable of maintaining vaginal pH and has no adverse effect on the beneficial vaginal flora. In still another embodiment of the present invention, wherein the composition show 100% non-motility in sperms. In still another embodiment of the present invention, wherein the composition show decrease in sperm hypo-osmotic swelling to about 8%. In still another embodiment of the present invention, wherein the composition show decrease in nuclear chromatin de-condensation to ranging between 25 to 30%. In still another embodiment of the present invention, wherein the composition show 100% contraceptive efficacy in first two hours of application. In still another embodiment of the present invention, wherein the composition show 77% contraceptive efficacy in first 24 hours of application. In still another embodiment of the present invention, wherein the composition show reversibility of contraceptive effect immediately after stopping the same. In an embodiment of the present invention, wherein a method of preparing a water based composition useful as vaginal contraceptive, said composition comprising hydrogen peroxide of concentration ranging between 1 to 3%, and sodium chloride of concentration ranging between 0.4 to 3.0%, with pH ranging between 4 to 4.5 and showing synergistic contraceptive effect, and said composition is an inexpensive, non-messy, fast acting vaginal contraceptive, with 100% efficacy and without any side effects. In another embodiment of the present invention, wherein adding sodium chloride to water. In yet another embodiment of the present invention, wherein obtaining sodium chloride solution of concentration ranging between 0.4 to 3%. In still another embodiment of the present invention, wherein mixing hydrogen peroxide to said sodium chloride solution. In still another embodiment of the present invention, wherein obtaining said mixture with hydrogen peroxide of concentration ranging between 1 to 3%. In still another embodiment of the present invention, wherein adjusting pH ranging between 4 to 4.5 In still another embodiment of the present invention, wherein obtaining said composition. In still another embodiment of the present invention, wherein the said solution has the capacity to increase the oxidative stress on the sperms limiting their progressive motility and affecting other sperm functions as well. In still another embodiment of the present invention, wherein isotonicity of the said composition would also be able to minimize any undesired effect of the hydrogen peroxide on the existing beneficial vaginal flora. In still another embodiment of the present invention, wherein chemical composition of the said vaginal contraceptive (i) NaCl (0.4 - 3.0%) in deionized triple distilled water, (ii) (1.0-3.0 %) H2O2, said composition at pH ranging between 4.0 to 4.5 In still another embodiment of the present invention, wherein volume of ejaculate varies with each ejaculate, thus effect of the test solution to varied volumes of semen is checked systematically for the loss of sperm motility over time. hi still another embodiment of the present invention, wherein as not all the non-motile sperms are necessarily non-viable, thus, viability testing provides information on the live and dead ratio of the sperm population, both motile and non-motile. In still another embodiment of the present invention, wherein testing spermicidal efficacy of any composition is thus important to find out the live and dead ratio of the sperms following an in vitro challenge. In still another embodiment of the present invention, wherein said composition is stored in dark bottles. In still another embodiment of the present invention, wherein one application of said composition is effective only during one intercourse. In still another embodiment of the present invention, wherein use of an inexpensive, non-messy, safe and fast acting aqueous synergistic spermicidal composition of claim 16 of 100% efficacy as vaginal contraceptive inn animals including humans, wherein the said composition is applied in the vaginal region. In still another embodiment of the present invention, wherein the composition starts functioning within 20 seconds of application. In still another embodiment of the present invention, wherein the applying volume of the composition is ranging between 10(4,1 to 5000(jl In still another embodiment of the present invention, wherein the composition is to be applied every time before intercourse. In still another embodiment of the present invention, wherein the composition shows 100% contraceptive efficacy in first two hours of application. In still another embodiment of the present invention, wherein a method of treating sperms with an inexpensive, non-messy, safe and fast acting aqueous synergistic spermicidal composition of claim 16 of 100% efficacy as vaginal contraceptive in animals including humans, said method comprising step of exposing sperms to the said composition. In still another embodiment of the present invention, wherein said method shows composition starting functioning within 20 seconds of application. In still another embodiment of the present invention, wherein applying volume of the said composition varies between 10(4,1 to 5000^1. In still another embodiment of the present invention, wherein said method producing composition showing decrease in sperm viability to about 10%. In still another embodiment of the present invention, wherein said method shows producing composition showing 100% non-motility in sperms. In still another embodiment of the present invention, wherein said method shows producing composition showing decrease in sperm hypo-osmotic swelling to about 8%. In still another embodiment of the present invention, wherein said method shows producing composition showing decrease in nuclear chromatin de-condensation to ranging between 25 to 30%. In still another embodiment of the present invention, wherein said method shows producing composition showing 100% contraceptive efficacy in first two hours of application. In still another embodiment of the present invention, wherein said method shows producing composition showing 77% contraceptive efficacy in first 24 hours of application. In still another embodiment of the present invention, wherein said method shows producing composition showing reversibility of contraceptive effect immediately after stopping the same. In still another embodiment of the present invention, wherein use of an inexpensive, non-messy, safe and fast acting aqueous synergistic spermicidal composition of claim 16 of 100% efficacy as vaginal contraceptive inn animals including humans, wherein the said composition is applied in the vaginal region. In still another embodiment of the present invention, wherein a method of treating sperms with an inexpensive, non-messy, safe and fast acting aqueous synergistic spermicidal composition of claim 16 of 100% efficacy as vaginal contraceptive in animals including humans, said method comprising step of exposing sperms to the said composition. A method of preparing an aqueous spermicidal composition, substantially as herein described with respect to the accompanying drawings and below-stated examples. A water based composition, substantially as herein described with respect to the accompanying drawings and below-stated examples. A method of preparing an aqueous spermicidal composition, substantially as herein described with respect to the accompanying drawings and aforesaid examples. A water based composition, substantially as herein described with respect to the accompanying drawings and aforesaid examples. The said composition is extremely economical and superior to commercially available vaginal contraceptive and can be of great use for a country like India. The details are as follows: Cost of commercially available drugs: (a) Deflon — 50 gms - cream -Rs. 10.00 approx. ( for one applications) (b) Orthognal - 85 gms - gelly - Rs. 4.00 approx. (for one applications) (c) Today - 5 pessaries -tablet -Rs. 14.00 approx. (for one application) (d) Applicants soln. - water based - Rs. 3-47- approx. (for about 100 applications) affecting the vaginal flora, affects vaginal pH maintains pH and provides contraceptive efficacy too Economy Minimum cost per application varies from Rs.4.00-10.00 Minimum cost per 100 application would be in the same range of Rs.3-4/-approx. Convenience Oil based, sexual pleasure is affected Water based therefore should not affect sexual pleasure Protection for HIV/AIDs Not reported FhCh is viricidal therefore might provide protection The components of the said composition are not capable of producing desired results individually. The synergistic contraceptive activity is shown by the said composition. The NaCl alone at concentration about 0.9% show 8% contraceptive activity, and hydrogen peroxide alone at concentration about 2% show 42% contraceptive activity. Surprisingly, both the said components put together shows synergistic contraceptive activity of 100% Brief description of the accompanying drawings Fig 1 shows sperm nuclear chromatin decondensation with about 70 % in control groups. Fig 2 shows after immobilization with test solution, a significantly (p Fig 3 shows viable sperms in the semen smear as cells with bright and colorless head, as viable cells do not pick color strain. (Control) Fig 4 shows dead sperms in the semen smear, as only dead cells pick up the strain after adding test solution. Fig 5 shows Control vagina with convoluted vaginal epithelium, which appeared normal with smooth boundaries. No inflammation of any kind was seen in any of the glandular cellular layers. The outer most epithelium lining was found a shade darker than the immediate innermost layer and could be differentiated from the other glandular cells. Fig 6 shows vaginal tissue after administering NaCl, with convoluted epithelium with inflammation at the outermost layer. As a result, the height of the cellular lining was found markedly increased. Fig 7 shows vaginal tissue after administering E^Oi with the outermost epithelium depicted less inflammatory but more pyknotic-staining pattern. This differential intense staining and contrast is mostly considered as apoptotic, inducing cell death in the affected region. This region can be differentiated from other glandular cells because of this typical staining pattern. The cells were more compressed and the cell height decreased. Increased vascularization in the innermost layers was noticed. Fig 8 shows vaginal tissue after administering test solution with differential staining of the innermost layer demarcated as was seen in control vagina. However, inflammatory changes in the innermost layer were still persisting similar to what has been observed in the vagina flushed with NaCl only. The significant improvement over raw H2O2 treatment was that there was no vascularization of tissue adjacent to the epithelial lining. The epithelial lining depicted restoration in the cellular height indicating normal secretary activity. There was no intense or pyknotic staining of the cellular epithelial lining. The following examples are given by the way of illustration of the present invention and should not be construed to limit the scope of the present invention. Examples: Example 1. - Sperm motility Semen and test solution was mixed in 1:1 proportion and a drop of the mixture was examined at 400 x under a phase contrast microscope to record the motility of the sperm in the preparation (WHO, 1999). Example 2: In vitro testing of human sperm motility with different dilutions of composition in semen Method: Effect of the Test Soln.[2% H2O2 in normal saline (0.9 % NaCl)] on Human Sperm Method: Eosin-Nigrosin staining (WHO, 1999) was utilized for recording sperm viability in control and test incubations. Briefly, 50 j^l of semen was properly mixed with 50 ^il of test solution. 150 ul Eosin Y (1.0 %).was added and after 30 seconds, three drops of Nigrosin (10 %) was added and mixed well. A smear on a clean glass slide was made from the mixture and allowed to air dry. Pink stained (dead) ones were differentiated from unstained, bluish fluorescent (live) sperms through examination using a phase contrast microscope. (As shown in figures 3 and 4) Fig 3 shows viable sperms in the semen smear as cells with bright and colorless head, as viable cells do not pick color strain. (Control). Fig 4 shows dead sperms in the semen smear, as only dead cells pick up the strain after adding test solution. Example 4. Hypo-osmotic swelling test Method: A hypo-osmotic swelling solution was prepared with sodium citrate 0.735 % and fructose 1.351 % in distilled water. Semen and the test solution was mixed in 1:1 proportion. 100 (j.1 of the sperm mixture was added in a pre-warmed 1-ml hypo-osmotic swelling solution and incubated at 37°C for 30 min. Swelling as indicated by various types of coiling of the sperm tail (in 100 random sperms) was examined using a phase contrast microscope (Jeyendran et al., 1984). Example 5: Nuclear chromatin de-condensation test Method: Spermatozoa pelleted from plasma or media by centrifugation at 400 g for 15 min. were resuspended in borate buffer (0.05 M) containing dithiotheritol (DTT, 2mM). Following incubation for 30 min at room temperature, 1 % lauryl sulfate (SDS) was added and mixed gently. The mixture was diluted with equal volume of borate buffer (0.05 M) containing 2.5 % glutaraldehyde. An aliquot of the mixture was stained with Rose Bengal (0.8 %) and was examined under a phase contrast microscope. The number of condensed and decondensed nuclear chromatin were recorded (Rodriguez et al., 1985). Sperm nuclear chromatin decondensation was 70 % in control groups (as shown in figure 1). Fig 1 shows sperm nuclear chromatin decondensation with about 70 % in control groups. However, after immobilization with H2O2, a significantly (p Example 6: pH of different test solution Method: pH was measured using a pH meter against a standard solution. pH level in different test solutions *pH in the vaginal cavity ranges from 4-4.5 Example 7. Tissue compatibility of the test solution in the vagina Cyto-toxicity to the epithelial lining is conventionally validated through the following observations: (a) Decrease in cell height confirms cyto-toxicity. (b) Shrinkage of the secretary glands confirms cyto-toxicity. (c) Increased vascularization confirms cyto-toxicity. (d) Pyknotic (thickened) epithelial cell margin confirms cyto-toxicity. After 24 hrs of administration vaginal tissue were processed for histology and analyzed under microscope. Fig 5 shows Control vagina with convoluted vaginal epithelium, which appeared normal with smooth boundaries. No inflammation of any kind was seen in any of the glandular cellular layers. The outer most epithelium lining was found a shade darker than the immediate innermost layer and could be differentiated from the other glandular cells. Fig 6 shows vaginal tissue after administering NaCl, with convoluted epithelium with inflammation at the outermost layer. As a result, the height of the cellular lining was found markedly increased. Fig 7 shows vaginal tissue after administering H2O2 with the outermost epithelium depicted less inflammatory but more pyknotic-staining pattern. This differential intense staining and contrast is mostly considered as apoptotic, inducing cell death in the affected region. This region can be differentiated from other glandular cells because of this typical staining pattern. The cells were more compressed and the cell height decreased. Increased vascularization in the innermost layers was noticed. Fig 8 shows vaginal tissue after administering test solution with differential staining of the innermost layer demarcated as was seen in control vagina. However, inflammatory changes in the innermost layer were still persisting similar to what has been observed in the vagina flushed with NaCl only. The significant improvement over raw H2O2 treatment was that there was no vascularization of tissue adjacent to the epithelial lining. The epithelial lining depicted restoration in the cellular height indicating normal secretary activity. There was no intense or pyknotic staining of the cellular epithelial lining. Table 7 showing the conditions of the vaginal epithelium following the administration of various solutions: Example 8: In vivo contraceptive efficacy testing of the chemical composition/test solution Method: Groups of rats in the proestrus and estrus stage of the cycle were vaginally administrated the test solutions at a specific volume as indicated below and immediately put to mating with a proven fertile male. After completion of specified time duration of co-habitation, mating was confirmed over the presence of spermatozoa in the vaginal swab analysis under microscope. The males were separated and the females were continued observation for 21 days till parturition for the final confirmation on the contraceptive efficacy of the test solution. Many a times in some rats the test solutions drop back following administration into the vaginal cavity once the rat is brought back to the normal dorsal position. This probably seems to be one of the problems in lower animals (rats) for testing a vaginal contraceptive. In contrast, potential space is available in the posterior fernix of the vagina in humans, which can hold back the contraceptive following administration. Example 9 Comparative contraceptive efficacy of test solution Method - Groups of rats in the proestrus and estrus stage of cycle were vaginally administered the test solutions at a specific volume as indicated below and immediately put to mating with a proven fertile male. After completion of specified time duration of co-habitation, mating was confirmed over the presence of spermatozoa in the vaginal swab analysis under microscope. The males were separated and the females were under continued observation for 21 days till parturition for the final confirmation on the contraceptive efficacy of the test solution. Example 10 Studies on reversibility of the contraceptive effect Method - groups of rates in the proestrus and estrus stage of the cycle were vaginally administered the test solutions at a specific volume for one time and for three consecutive cycles below and immediately put to mating with a proven fertile male (1:1); After continuation of the efficacy in the three consecutive cycles and after 21 days (gestation period in arts), the rats were not administered the contraceptive test solution but allowed to mate with the male. The females were again watched till parturition (21 days) for confirmation of pregnancy and the size of the litters. The litters do not show any abnormality after administering the test solution of 0.9% NaCl and about 2 % hydrogen peroxide. Result: females not administered the composition of the instant Application became pregnant thus confirming the reversibility of the composition. We claim 1. A water based spermicidal vaginal contraceptive composition comprising (i) hydrogen peroxide of concentration ranging from 1 to 3%, and (ii) sodium chloride of concentration ranging between 0.4 to 3%, with pH ranging between 4 to 4.5 showing synergistic contraceptive effect. 2. A composition as claimed in claim 1, wherein said composition preferred concentration of hydrogen peroxide is 2%. 3. A composition as claimed in claim 1, wherein said composition preferred pH is 4.15. 4. A composition as claimed in claim 1, wherein application volume of the said composition varies between l0µl to 5000µl. 5. A composition as claimed in claim 1, wherein said composition is most effective in the semen volume in the ratio ranging between 1:1 to 1:5. 6. A composition as claimed in claim 1, wherein said composition shows decrease in sperm viability to about 10%. 7. A composition as claimed in claim 1, wherein said composition shows 100% non¬motility in sperms. 8. A composition as claimed in claim 1, wherein said composition shows decrease in sperm hypo-osmotic swelling to about 8%. 9. A composition as claimed in claim 1, wherein said composition shows decrease in nuclear chromatin de-condensation to ranging between 25 to 30%. 10. A composition as claimed in claim 1, wherein said composition shows 100% contraceptive efficacy in first two hours of application. 11. A composition as claimed in claim 1, wherein said composition shows 77% contraceptive efficacy in first 24 hours of application. 12. A composition as claimed in claim 1, wherein said composition shows reversibility of contraceptive effect immediately after stopping the same. 13. A method of preparing an inexpensive, non-messy, safe and fast acting aqueous synergistic spermicidal composition of claim 1 with 100% efficacy useful as a vaginal contraceptive free of side effects, said method comprising steps of; (i) preparing sodium chloride solution of concentration ranging between 0.4 to 3% in water, (ii) mixing hydrogen peroxide to said sodium chloride solution to obtain a mixture having hydrogen peroxide in the concentration ranging between 1 to 3%, (iii) adjusting pH of the mixture in the range between 4 to 4.5 by altering the volume of sodium chloride and hydrogen peroxide, and (iv) obtaining said composition. 14. A method as claimed in claim 13, wherein said composition shows preferred concentration of hydrogen peroxide of about 2%. 15. A method as claimed in claim 13, wherein said composition shows preferred pH of about 4.15. 16. A method as claimed in claim 13, wherein said composition have no adverse effect on the vagina. 17. A method as claimed in claim 13, wherein said method composition have a shelf life of about 30 days. 18. A method as claimed in claim 13, wherein said composition is safe for application in animals including humans. 19. A method as claimed in claim 13, wherein said method shows composition starting functioning within 20 seconds of application. 20. A method as claimed in claim 13, wherein applying volume of the said composition varies between 10µ1 to 5000µl. 21. A method as claimed in claim 13, wherein said method producing composition showing decrease in sperm viability to about 10%. 22. A method as claimed in claim 13, wherein said method shows producing composition showing 100% non-motility in sperms. 23. A method as claimed in claim 13, wherein said method shows producing composition showing decrease in sperm hypo-osmotic swelling to about 8%. 24. A method as claimed in claim 13, wherein said method decrease nuclear chromatin de-condensation to the range of 25 to 30%. 25. A method as claimed in claim 13, wherein said composition show 100% contraceptive efficacy in first two hours of application. 26. A method as claimed in claim 13, wherein said composition show 77% contraceptive efficacy in first 24 hours of application. 27. A method as claimed in claim 13, wherein said method shows producing composition showing reversibility of contraceptive effect immediately after stopping the same. 28. A water based spermicidal vaginal contraceptive composition, a method of preparing an inexpensive, non-messy, safe and fast acting aqueous synergistic spermicidal composition of 100% efficacy useful as a vaginal contraceptive free of side effects substantially as herein described and illustrated therein.

Documents:

07-del-2002-abstract.pdf

07-del-2002-claims.pdf

07-del-2002-correspondence-others.pdf

07-del-2002-description (complete)-04-09-2008.pdf

07-del-2002-description (complete)-18-06-2008.pdf

07-del-2002-description (complete)-21-05-2008.pdf

07-del-2002-description (complete).pdf

07-del-2002-drawings.pdf

07-del-2002-form-1.pdf

07-del-2002-form-18.pdf

07-del-2002-form-2.pdf

07-del-2002-form-26.pdf

07-del-2002-form-3.pdf

07-del-2002-form-4.pdf

07-del-2002-form-5.pdf

7-DEL-2002-Abstract-(18-06-2008).pdf

7-DEL-2002-Abstract-21-05-2008.pdf

7-DEL-2002-Claims-(04-09-2008).pdf

7-DEL-2002-Claims-(18-06-2008).pdf

7-DEL-2002-Claims-21-05-2008.pdf

7-DEL-2002-Correspondence-Others-(04-09-2008).pdf

7-DEL-2002-Correspondence-Others-(18-06-2008).pdf

7-DEL-2002-Correspondence-Others-21-05-2008.pdf

7-DEL-2002-Form-1-(18-06-2008).pdf

7-DEL-2002-Form-1-21-05-2008.pdf

7-del-2002-form-13-(16-09-2008).pdf

7-del-2002-form-13-(21-05-2008).pdf

7-DEL-2002-Form-2-21-05-2008.pdf

7-DEL-2002-Petition-137-21-05-2008.pdf