[0001] The present invention relates to a mosquito repellent composition. Specifically, the present invention relates to a mosquito repellent composition, comprising natural ingredients and having cleansing and anti-microbial properties.

BACKGROUND OF THE INVENTION
[0002] Background description includes information that may be useful in understanding the present invention. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, or that any publication specifically or implicitly referenced is prior art.
[0003] Vector-borne diseases have become a global health problem. Using appropriate insect repellents to prevent the mosquito from biting the human or animal is the most logical approach to suppressing these vector-borne diseases. Repellents make humans unattractive to biting flies, such as a mosquito, so that the biting fly will avoid areas of the body that have been treated with the product. Repellents do not kill biting flies, such as mosquitoes. The best repellents will provide protection from bites for a long period of time from just one application.
[0004] The mosquito-borne diseases are on rise in many developing countries globally. The mosquito-borne illness, malaria, is one of the main killer diseases of the world. Although malaria can be cured with prescription drugs, many mosquito species have developed resistance to common anti-malarial drugs. The clinical need to maximise protection in the world zones experiencing serious disease including Malaria, Dengue fever and Zika requires an urgent search for more effective repellents offering greater longevity of protection. The Aedes aegypti mosquito, originally from Africa, is known to spread yellow fever, dengue and dengue hemorrhagic fever. The Aedes albopictus mosquito, from Asia, is known to spread several types of equine encephalitis virus, dengue, dengue hemorrhagic fever, and recently, canine heartworm. In terms of Dengue fever and the Zika virus, the Aedes Aegypti mosquito vector is unusually a "day biter", so the resulting requirement for 12 hours protection is an actual pre-requisite for an effective insect repellent for this species.
[0005] Insect repellents that are topically applied to deter mosquitoes from biting consist of chemical compositions or essential oils. The commercial mosquito repellent coils and vaporizers use toxic chemical agents such as dimethyl phthalate and N,N-diethyl-m-toluamide (DEET), which have been found to be associated with rashes, skin irritation, numb or burning lips, nausea, headaches, dizziness and difficulty concentrating; and, further worse, behavioral changes and even diffuse brain cell death - on frequent and prolonged exposure.
[0006] The currently accepted first line of defence agreed by all healthcare/regulatory bodies aimed at disease prevention is more effective repellency. There is a need for compositions that show high levels of efficacy against different species of mosquito vector and can offer protection for periods of hours consistently in excess of current standards.
[0007] There is therefore, a need in the art for safer and more effective mosquito repellents that are devoid of, or at least alleviates, the disadvantages associated with known mosquito repellents. Need is also felt for mosquito repellent formulation that does not contain harmful chemicals, is safe and nontoxic. The new advanced mosquito repellent as disclosed in the present disclosure offers natural protection from mosquito born disease while its unique formulation creates a shield around the area on which mosquito repellent is applied, and provides ultimate protection even from the dreaded disease causing mosquitoes.

OBJECTS OF THE INVENTION
[0008] An object of the present disclosure is to provide a mosquito repellent composition which satisfies the existing needs, as well as emerging ones, and generally overcomes the deficiencies found in the prior art.
[0009] Another object of the present invention is to provide a mosquito repellent composition, comprising natural ingredients, which is safe, effective and devoid of side effects.
[0010] Another object of the present invention is to provide a mosquito repellent composition that does not elicit any skin reactions.
[0011] Another object of the present invention is to provide a non-irritating and dermatologically safe mosquito repellent composition.
[0012] Another object of the present invention is to provide complete protection time (CPT) of the mosquito repellent composition.
[0013] Another object of the present invention is to provide a mosquito repellent composition, which is easy to use and provides long term protection.
[0014] Another object of the present invention is to provide a mosquito repellent composition, which has pleasant fragrance, and can be used indoors as well as outdoors.
[0015] Another object of the present invention is to provide a mosquito repellent composition, which alleviates the danger of diseases like Zika Fever, Dengue, Chikungunya, Malaria and Yellow fever.
[0016] Another object of the present invention is to provide a mosquito repellent composition, having cleansing and antimicrobial properties.
[0017] Another object of the present invention is to provide a mosquito repellent composition that provides antimicrobial efficacy of 99.99% against skin pathogenic bacteria like Staphylococcus aureus, Pseudomonas aeruginosa, Staphylococcus epidermidis and Candida albanicus.

SUMMARY OF THE INVENTION
[0018] The present invention relates to a mosquito repellent composition. Specifically, the present invention relates to a mosquito repellent composition, comprising natural active ingredients which is safe, effective and devoid of side effects.
[0019] In one aspect, the present invention relates to a mosquito repellent composition for repelling mosquitoes.
[0020] In one aspect, the present invention relates to a mosquito repellent composition comprising natural surfactant, natural oils, plant extracts, skin conditioner, emulsifiers, preservative, pH adjuster and anti-microbial agent, wherein natural oils and plant extracts act as natural repellent.
[0021] In one aspect, the present invention relates to a mosquito repellent composition wherein the natural surfactant used is selected from Rhamnolipid or Sodium coco sulpahte, preferably Rhamnolipid.
[0022] In one aspect, the present invention relates to a mosquito repellent composition wherein natural oils and plant extracts have mosquito repellent properties and act as anti-microbials.
[0023] In one aspect, the present invention relates to a mosquito repellent composition wherein natural oils and plant extracts are selected from Peppermint Oil, citronella oil, Lemon eucalyptus Oil, Tea Tree Oil, Calendula Oil, and the like or combinations thereof.
[0024] In one aspect, the present invention relates to a mosquito repellent composition wherein skin conditioner can be selected from Thiamine Hydrochloride, Calendula Oil and the like or combinations thereof.
[0025] In one aspect, the present invention relates to a mosquito repellent composition wherein Thiamine hydrochloride acts as a skin conditioner and as mosquito repellent.
[0026] In another aspect, the present invention relates to a mosquito repellent composition wherein natural preservative can be selected from Nisin, dehydroacetic acid and the like or combinations thereof.
[0027] In one aspect, the present invention relates to a mosquito repellent composition wherein Nisin acts as a natural preservative and anti-microbial agent.
[0028] In another aspect, the present invention relates to a mosquito repellent composition wherein emulsifier is selected from lecithin, sunflower lecithin and the like.
[0029] In one aspect, the present invention relates to a mosquito repellent composition having cleansing and anti-microbial properties.
[0030] In one aspect, the present invention relates to a mosquito repellent composition comprising :
a) Rhamnolipids ;
b) Nisin;
c) Peppermint Oil;
d) Lemon eucalyptus Oil ;
e) Citronella Oil;
f) Tea tree Oil;
g) Calendula Oil ;
h) Thiamine hydrochloride;
i) Dehydroacetic acid;
j) Sunflower Lecithin;
k) Sodium Hydroxide; and
l) Distilled water.
[0031] In another aspect, the mosquito repellent composition of the present invention comprises natural surfactant in an amount ranging from about 0.01% to about 5% by weight of the composition.
[0032] In another aspect, the mosquito repellent composition of the present invention comprises natural oils in an amount ranging from about 5% to about 40% by weight of the composition.
[0033] In another aspect, the mosquito repellent composition of the present invention comprises preservative in an amount ranging from about 0.001% to about 1% by weight of the composition.
[0034] In another aspect, the mosquito repellent composition of the present invention comprises skin conditioner in an amount ranging from about 0.1% to about 20% by weight of the composition.
[0035] In another aspect, the mosquito repellent composition of the present invention comprises emulsifier in an amount ranging from about 1% to about 10% by weight of the composition.
[0036] In another aspect, the mosquito repellent composition of the present invention comprises anti-microbial agents in an amount ranging from about 0.001% to about 25% by weight of the composition.
[0037] In one aspect, the present invention relates to a mosquito repellent composition comprising :
a) Rhamnolipids 0.1g;
b) Nisin 0.025g;
c) Peppermint Oil 2ml;
d) Lemon eucalyptus Oil 5ml;
e) Citronella Oil 5ml;
f) Tea tree Oil 10ml;
g) Calendula Oil 15ml;
h) Thiamine hydrochloride 0.1g;
i) Dehydroacetic acid 0.03g;
j) Sunflower Lecithin 4ml;
k) Sodium Hydroxide: Adjusted using 10M Stock solution; and
l) Distilled water q.s. to make 100mL.
[0038] In one aspect, the present invention relates to a mosquito repellent composition which is easy to use and provides long term protection upto 4-10 hours.
[0039] In another aspect, the present invention relates to a mosquito repellent composition that can be easily applied to the skin.
[0040] In one aspect, the present invention relates to a mosquito repellent composition, which can be formulated as a deodorizer, waterless body bath, cream, lotion or oil.
[0041] In one aspect, the present invention relates to a mosquito repellent composition, which is effective in repelling mosquitoes indoors as well as outdoors.
[0042] In one aspect, the present invention relates to a mosquito repellent composition which has pleasant fragrance.
[0043] In one aspect, the present invention relates to a mosquito repellent composition, which effectively repels mosquitoes that may transmit the Chikungunya, Zika, Dengue and West Nile viruses.
[0044] Various objects, features, aspects and advantages of the inventive subject matter will become more apparent from the following detailed description of preferred embodiments

DETAILED DESCRIPTION
[0045] The following is a detailed description of embodiments of the disclosure. The embodiments are in such detail as to clearly communicate the disclosure. However, the amount of detail offered is not intended to limit the anticipated variations of embodiments; on the contrary, the intention is to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the present disclosure as defined by the appended claims.
[0046] All publications herein are incorporated by reference to the same extent as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference. Where a definition or use of a term in an incorporated reference is inconsistent or contrary to the definition of that term provided herein, the definition of that term provided herein applies and the definition of that term in the reference does not apply.
[0047] Reference throughout this specification to “one embodiment” or “an embodiment” means that a particular feature, structure or characteristic described in connection with the embodiment is included in at least one embodiment. Thus, the appearances of the phrases “in one embodiment” or “in an embodiment” in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.
[0048] In some embodiments, the numbers expressing quantities of ingredients, properties such as concentration, reaction conditions, and so forth, used to describe and claim certain embodiments of the invention are to be understood as being modified in some instances by the term “about.” Accordingly, in some embodiments, the numerical parameters set forth in the written description and attached claims are approximations that can vary depending upon the desired properties sought to be obtained by a particular embodiment. In some embodiments, the numerical parameters should be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of some embodiments of the invention are approximations, the numerical values set forth in the specific examples are reported as precisely as practicable. The numerical values presented in some embodiments of the invention may contain certain errors necessarily resulting from the standard deviation found in their respective testing measurements.
[0049] As used in the description herein and throughout the claims that follow, the meaning of “a,” “an,” and “the” includes plural reference unless the context clearly dictates otherwise. Also, as used in the description herein, the meaning of “in” includes “in” and “on” unless the context clearly dictates otherwise.
[0050] Unless the context requires otherwise, throughout the specification which follow, the word “comprise” and variations thereof, such as, “comprises” and “comprising” are to be construed in an open, inclusive sense that is as “including, but not limited to.”
[0051] The recitation of ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range. Unless otherwise indicated herein, each individual value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g. “such as”) provided with respect to certain embodiments herein is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention otherwise claimed. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the invention.
[0052] Groupings of alternative elements or embodiments of the invention disclosed herein are not to be construed as limitations. Each group member can be referred to and claimed individually or in any combination with other members of the group or other elements found herein. One or more members of a group can be included in, or deleted from, a group for reasons of convenience and/or patentability. When any such inclusion or deletion occurs, the specification is herein deemed to contain the group as modified thus fulfilling the written description of all Markush groups used in the appended claims.
[0053] The description that follows, and the embodiments described therein, is provided by way of illustration of an example, or examples, of particular embodiments of the principles and aspects of the present disclosure. These examples are provided for the purposes of explanation, and not of limitation, of those principles and of the disclosure.
[0054] It should also be appreciated that the present disclosure can be implemented in numerous ways, including as a system, a method or a device. In this specification, these implementations, or any other form that the invention may take, may be referred to as processes. In general, the order of the steps of the disclosed processes may be altered within the scope of the invention.
[0055] The headings and abstract of the invention provided herein are for convenience only and do not interpret the scope or meaning of the embodiments.
[0056] The following discussion provides many example embodiments of the inventive subject matter. Although each embodiment represents a single combination of inventive elements, the inventive subject matter is considered to include all possible combinations of the disclosed elements. Thus if one embodiment comprises elements A, B, and C, and a second embodiment comprises elements B and D, then the inventive subject matter is also considered to include other remaining combinations of A, B, C, or D, even if not explicitly disclosed.
[0057] Various terms as used herein are shown below. To the extent a term used in a claim is not defined below, it should be given the broadest definition persons in the pertinent art have given that term as reflected in printed publications and issued patents at the time of filing.
[0058] The present invention relates to a mosquito repellent composition. Specifically, the present invention relates to a mosquito repellent composition, comprising natural active ingredients which is safe, effective and devoid of side effects.
[0059] In one embodiment, the present invention relates to a mosquito repellent composition for repelling mosquitoes.
[0060] In one embodiment, the present invention relates to a mosquito repellent composition comprising natural surfactant, natural oils and plant extracts, skin conditioner, emulsifiers, preservative, pH adjuster and anti-microbial agent, wherein natural oils and plant extracts act as natural repellent.
[0061] In one embodiment, the present invention relates to a mosquito repellent composition wherein the natural surfactant used is selected from Rhamnolipid or Sodium coco sulpahte, preferably Rhamnolipid.
[0062] In one embodiment, the present invention relates to a mosquito repellent composition wherein natural oils and plant extracts have mosquito repellent properties and act as anti-microbials.
[0063] In one embodiment, the present invention relates to a mosquito repellent composition wherein natural oils and plant extracts are selected from Peppermint Oil, citronella oil, Lemon eucalyptus Oil, Tea Tree Oil, Calendula Oil, and the like or combinations thereof.
[0064] Natural oils also referred to as essential oils are defined as any class of volatile oils obtained from plants possessing the odour and other characteristic properties of the plant. Examples of repellent compounds that are essential oils includes: almond bitter oil, anise oil, basil oil, bay oil, caraway oil, cardamom oil, cedar oil, celery oil, calendula oil, chamomile oil, cinnamon oil, citronella oil, clove oil, coriander oil, cumin oil, dill oil, eucalyptus oil, fennel oil, ginger oil, grapefruit oil, lemon oil, lime oil, lemon-eucalyptus oil, mint oil, parsley oil, peppermint oil, pepper oil, rose oil, spearmint oil (menthol), sweet orange oil, thyme oil, turmeric oil, tea tree oil and oil of wintergreen. Examples of active ingredients in essential oils are: citronella, methyl salicylate, ethyl salicylate, propyl salicylate, citronellol, safrole, and limonene.
[0065] In one embodiment, the present invention relates to a mosquito repellent composition wherein skin conditioner can be selected from Thiamine Hydrochloride, Calendula Oil and the like or combinations thereof.
[0066] In one embodiment, the present invention relates to a mosquito repellent composition wherein Thiamine hydrochloride acts as a skin conditioner and as mosquito repellent.
[0067] In another embodiment, the present invention relates to a mosquito repellent composition wherein preservative can be selected from Nisin, dehydroacetic acid and the like or combinations thereof.
[0068] In one embodiment, the present invention relates to a mosquito repellent composition wherein Nisin acts as a natural preservative and anti-microbial agent.
[0069] In another embodiment, the present invention relates to a mosquito repellent composition wherein emulsifier is selected from lecithin, sunflower lecithin and the like.
[0070] In one embodiment, the present invention relates to a mosquito repellent composition having cleansing and anti-microbial properties.
[0071] In one embodiment, the present invention relates to a mosquito repellent composition comprising :
a) Rhamnolipids ;
b) Nisin;
c) Peppermint Oil;
d) Lemon eucalyptus Oil ;
e) Citronella Oil;
f) Tea tree Oil;
g) Calendula Oil ;
h) Thiamine hydrochloride;
i) Dehydroacetic acid;
j) Sunflower Lecithin;
k) Sodium Hydroxide; and
l) Distilled water.
[0072] In another embodiment, the mosquito repellent composition of the present invention comprises natural surfactant in an amount ranging from about 0.01% to about 5% by weight of the composition, preferably in the amount ranging from about 0.05 % to about 3 % by weight of the composition.
[0073] In another embodiment, the mosquito repellent composition of the present invention comprises natural oils in an amount ranging from about 5% to about 50% by weight of the composition, preferably in an amount ranging from about 25% to about 35% by weight of the composition.
[0074] In another embodiment, the mosquito repellent composition of the present invention comprises Peppermint Oil in an amount ranging from about 0.5% to about 10%, preferably in the amount ranging from about 1 % to about 5 % by weight of the composition.
[0075] In another embodiment, the mosquito repellent composition of the present invention comprises lemon-eucalyptus oil in an amount ranging from about 1% to about 10%, preferably in the amount ranging from about 3 % to about 7 % by weight of the composition.
[0076] In another embodiment, the mosquito repellent composition of the present invention comprises Citronella Oil in an amount ranging from about 1% to about 10%, preferably in the amount ranging from about 3 % to about 7 % by weight of the composition.
[0077] In another embodiment, the mosquito repellent composition of the present invention comprises tea tree oil in an amount ranging from about 2% to about 20% preferably in the amount ranging from about 5 % to about 15 % by weight of the composition.
[0078] In another embodiment, the mosquito repellent composition of the present invention comprises calendula oil in an amount ranging from about 5% to about 30% preferably in the amount ranging from about 10 % to about 20 % by weight of the composition.
[0079] In another embodiment, the mosquito repellent composition of the present invention comprises preservative in an amount ranging from about 0.001% to about 1% by weight of the composition, preferably in an amount ranging from about 0.005% to about 0.5% by weight of the composition.
[0080] In another embodiment, the mosquito repellent composition of the present invention comprises skin conditioner in an amount ranging from about 0.1% to about 20% by weight of the composition, preferably in an amount ranging from about 0.5 % to about 15% by weight of the composition.
[0081] In another embodiment, the mosquito repellent composition of the present invention comprises emulsifier in an amount ranging from about 0.5% to about 10% by weight of the composition, preferably in an amount ranging from about 1% to about 5% by weight of the composition.
[0082] In another embodiment, the mosquito repellent composition of the present invention comprises anti-microbial agents in an amount ranging from about 0.001% to about 25% by weight of the composition, preferably in an amount ranging from about 0.01% to about 20% by weight of the composition.
[0083] In one embodiment, the present invention relates to a mosquito repellent composition comprising :
a) Rhamnolipids 0.1g;
b) Nisin 0.025g;
c) Peppermint Oil 2ml;
d) Lemon eucalyptus Oil 5ml;
e) Citronella Oil 5ml;
f) Tea tree Oil 10ml;
g) Calendula Oil 15ml;
h) Thiamine hydrochloride 0.1g;
i) Dehydroacetic acid 0.03g;
j) Sunflower Lecithin 4ml;
k) Sodium Hydroxide: Adjusted using 10M Stock solution; and
l) Distilled water q.s. to make 100mL.
[0084] In one embodiment, the present invention relates to a mosquito repellent composition which is easy to use and provides long term protection upto 4-10 hours.
[0085] In another embodiment, the present invention relates to a mosquito repellent composition which can be easily applied on the skin.
[0086] In one embodiment, the present invention relates to a mosquito repellent composition, which can be formulated as a deodorizer, waterless bodybath, cream, lotion or oil.
[0087] In one embodiment, the present invention relates to a mosquito repellent composition, which cleanses the skin and provides anti-microbial protection when used as a bodybath.
[0088] In one embodiment, the present invention relates to a mosquito repellent composition, which is effective in repelling mosquitoes indoors as well as outdoors.
[0089] In one embodiment, the present invention relates to a mosquito repellent composition which has pleasant fragrance.
[0090] In one embodiment, the present invention relates to a mosquito repellent composition, which effectively repels mosquitoes that may transmit the Chikungunya, Zika, Dengue and West Nile viruses.
[0091] In another embodiment, the present invention relates to a mosquito repellent composition, which is safe for use by anyone, even children and elderly, without any harmful side effects.
[0092] The inventors of the present invention have surprisingly found that the efficacy of the present invention and synergistic effect of the composition lies in the formulation of the composition of the ingredients in a particular ratio which is highly significant. The composition of the present invention not only repels mosquitoes but exhibits anti-microbial properties in addition to being safe for use on skin for long periods of time. The composition of the present invention has cleansing properties due to which it can be used as anti-mosquito body bath.
[0093] According to another embodiment of the present invention, the mosquito repellent composition was tested for Skin Irritation on human subjects. The study has been conducted as per “IS 13424: 2001 (Reaffirmed 2006)” and the composition was found to be “safe for human skin”. In other words, the mosquito repellent composition according to the embodiments of the present invention is dermatologically tested and found to be safe. The details of the study are reported in Example 1.
[0094] According to another embodiment of the present invention, the mosquito repellent composition was tested for antimicrobial efficacy against skin pathogenic bacteria The results, as depicted in Example 2 and Table 1 indicate that antimicrobial efficacy of composition of the present invention was found to be 99.99% effective against bacterial cultures Staphylococcus aureus, Pseudomonas aeruginosa, Staphylococcus epidermidis and fungal culture Candida albanicus.
[0095] According to another embodiment of the present invention, complete protection time (CPT) was evaluated for the mosquito repellent composition, in comparison with DEET (N,N-diethyl-3-methylbenzamide; Positive control, used in 100% concentration as positive control, known to be toxic). The composition according to the embodiments of the present invention was applied on human volunteer’s skin and was tested against female mosquito species Anopheles stephensi. CPT was found to be 240 ± 1.96*34.35 minutes for the composition according to present invention and 360 ± 1.96*35.81 minutes for positive control. On this basis, in 95 out of 100 observations, median CPT of the composition of the present invention lies between 173 to 307 minutes and that of positive control DEET lies between 290 to 430 minutes. The details of the study are reported in Example 3.
[0096] In one embodiment, the present invention relates to a mosquito repellent composition that did not induce any skin sensitization (allergies) to the skin of the guinea pigs when compared to its corresponding control group of animals. The study has been conducted as per IS 13424: 2001 (Reaffirmed 2006). The details of the study are reported in Example 4.
[0097] While the foregoing describes various embodiments of the disclosure, other and further embodiments of the disclosure may be devised without departing from the basic scope thereof. The scope of the invention is determined by the claims that follow. The invention is not limited to the described embodiments, versions or examples, which are included to enable a person having ordinary skill in the art to make and use the invention when combined with information and knowledge available to the person having ordinary skill in the art.
[0098] The present invention is further explained in the form of following examples. However, it is to be understood that the following examples are merely illustrative and are not to be taken as limitations upon the scope of the invention.
[0099] All the starting materials are commercially available and were purchased from PK Scientific Pvt. Ltd. Company, Delhi. The studies on the efficacy and safety of the composition were carried out at Toxicology Centre Shriram Institute for Industrial Research, University Road, Delhi.
[00100] Example 1: Skin Irritation (Patch Test) Study
[00101] This study was conducted based on the requirements of IS 13424: 2001 (Reaffirmed 2006) and was designed to determine skin irritation of the composition of the present invention, after a single application (patch test) in the human subjects.
[00102] In the study, 15 males and 15 females of good health or physical condition were enrolled. The composition of the present invention was diluted in distilled water (8%) & used for application. Distilled water was used as negative control and 3 % Sodium Lauryl Sulphate (SLS) (3 gm of SLS was dissolved in 100ml distilled water to make it a clear, transparent solution) was used as the positive control.
[00103] On the day of application, 40µl of 8 % of solution of composition of the present invention was gently applied on the inner aspect of the upper arm at the site marked as “3”.For comparison 40µl of distilled water (negative control) was also dispensed in separate Finn chambers and gently applied on the inner aspect of the upper arm at the site marked as “1”. Similarly, 40µl of 3% (w/v) aqueous solution of Sodium Lauryl Sulphate (positive control) was also dispensed in separate Finn chambers and applied on the inner aspect of the upper arm at the site “2”. The patches were held in place by securing it with a non-allergic tape. After 24 hours of the exposure period, the patches were removed gently and the sites were cleared with luke warm water to remove the left over test items and marked for identification. The reactions were observed at 24 hours, 72 hours, and 7 days after the patch removal. Under the conditions of this study, the single application of composition of the present invention did not elicit any skin reactions. The mean score for composition of the present invention was 0.20 at 24 hours after patch removal.
[00104] Example 2: Antimicrobial Efficacy against Skin Pathogenic Bacteria
[00105] This study was conducted as per the guidelines ASTM E 2315: Standard guide for Assessment of Antimicrobial activity using a time kill procedure.
[00106] Stock spore suspensions of test organisms were prepared. Bacterial cultures were prepared for Staphylococcus aureus, Pseudomonas aeruginosa and Staphylococcus epidermidis as to obtain a count of at least 108 cfu per ml by using densitometer. Fungal culture of Candida albicans was prepared as to obtain a count of at least 108 cfu per ml by using densitometer.
[00107] TEST PROCEDURE: 10 ml of the sample was prepared with respect to individual test culture. 100 µl of adjusted 106 cfu/ml of the test cultures were inoculated in individual sample prepared. A contact time of 3 minutes was taken in consideration in the test. Thus a ‘0’ min and ‘3’ was taken. Plates were incubated at 30°C for 48 hrs. The Count was taken and observations recorded. For population quantitation of the control blank: A volume of inoculum suspension was also inoculated into the 10ml of dilution blank (saline).The initial and final count of the population in the blank was done for valid test. The results are depicted in Table 1.
Table 1: Efficacy of the composition of present invention against microorganisms:
Challenged
Microorganisms Count at 0 min
(in cfu)
Count after 3 min
(in cfu)
% Reduction

Staphylococcus aureus 2.8 x 106 No colony observed 99.99
Staphylococcus epidermidis 3.2 x 106 No colony observed 99.99
Pseudomonas aeruginosa 4.1 x 106 No colony observed 99.99
Candida albicans 2.4 x 106 No colony observed 99.99
[00108] The results indicate that antimicrobial efficacy of composition of the present invention was found to be 99.99% effective against bacterial cultures Staphylococcus aureus, Pseudomonas aeruginosa, Staphylococcus epidermidis and fungal culture Candida albanicus.
[00109] Example 3: Mosquito Repellency activity and Complete Protection Time on Human Skin
[00110] The study was conducted to meet the requirement of WHO Guidelines for Efficacy Testing of Mosquito Repellents for Human Skin, 2009. 40 Healthy (20 male and 20 female), volunteers of age 18-60 years were selected for the study. The volunteers were exposed to the 1 ml of the 20 % ethanolic DEET solution on one arm and compared with same amount of the composition of the present invention, on other arm for 8 hours.
[00111] Mosquito repellency test was conducted with female mosquitoes species Anopheles stephensi.1 ml of the 20% ethanolic DEET solution (N,N-diethyl-meta-toluamide) was used as positive control and compared with 1mL of the composition of the present invention. Both were applied to the area of 600 cm2 of the forearm skin between the wrist and elbow. The 40 healthy individuals were observed after every hour for 3 minutes. Complete protection time was calculated as the number of minutes elapsed between the time of repellent application and the first mosquito landing and/or probing. Repellency activity study was completed in 8 hours.
[00112] Median and Confidence Interval of Complete Protection Time were estimated using the Kaplan Meier Survivor Function. The estimated median CPT is the smallest observed CPT for which S(t)=0.5.The 95 % confidence interval of this estimate is given by t(50)±1.96.
[00113] Result of the studies indicate that under the conditions of this study, the 95 % confidence interval for the estimated median Complete Protection Time (CPT) for the composition of the present invention and DEET (N,N-diethyl-meta-toluamide, Positive control) applied on Human volunteer’s skin against female mosquito species Anopheles stephensi was found to be 240 ± 1.96*34.35 minutes and 360 ± 1.96*35.81. On this basis, in 95 out of 100 observations, median CPT of the composition of the present invention lies between 173 to 307 minutes and that of positive control DEET lies between 290 to 430 minutes.
[00114] Example 4: Skin Sensitization Study (Buehler Test) in Dunken Hartley Guinea Pigs
[00115] The study was conducted to meet the requirement of IS 13424: 2001 (Reaffirmed 2006). This study was designed to determine the skin sensitization potential of the composition of the present invention on contact with Skin of Dunken Hartley Guinea Pigs, following the method described by Buehler (1965).
[00116] Initially, a pilot study was conducted for titrating suitable dose for the induction and challenge exposure. A volume of 0.5 ml each of 4 serial concentrations 25, 50, 75 and 100 % of composition of the present invention were applied on the back of four guinea pigs and the test site were observed after 6 hours for any skin reaction. Since, 0.5 ml, the maximum recommended dose, did not revealed any signs for cutaneous/ general toxicity in guinea pig in the pilot study, thus, 100% dose concentration was selected for induction and challenge phases respectively in the main study.
[00117] In the main test, thirty animals were randomly selected and divided in two groups. 1st group i.e. control group consisted five male and five female animals and 2nd group i.e. treated group consisted of Ten Male and Ten Female animals. 24 hrs before the test the hair on the back of each guinea pig were closely clipped on the both side of the spinal column. The composition of the present invention was evenly applied on the left side of clipped back at the dose level of 0.5 ml of 100 % of test item per treated animal and control group animals applied with gauze patch only during induction phase. The test item was applied on day 1, 8 and 15 days respectively on test animals during the induction (sensitization) phase and Challenge exposure was made after completion of fourteen days from last exposure both in treated and control groups. After 24 and 48 hours of patch removal in the challenge phase all the animals were observed in light for any reaction and skin of test sites was graded as per Magnusson & Kligman Scale.
[00118] No gross skin reactions (erythema or edema) were observed at the test sites and in the area surrounding it in any the guinea pigs of control and treated groups. Mean body weights of treated guinea pigs were also comparable to control group animals. Therefore, under the conditions of this study, the repeated application of composition of the present invention did not induce skin sensitization (allergies) in any of the test guinea pigs, when compared to its corresponding control group of animals.
[00119] The foregoing examples are merely illustrative and are not to be taken as limitations upon the scope of the invention. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art. Such changes and modifications may be made without departing from the scope of the invention.

ADVANTAGES OF THE PRESENT INVENTION
[00120] The present invention provides a mosquito repellent composition, comprising natural ingredients, which is safe, effective and devoid of side effects.
[00121] The present invention provides a mosquito repellent composition that does not elicit any skin reactions.
[00122] The present invention provides a non-irritating and dermatologically safe mosquito repellent composition.
[00123] The present invention provides a complete protection time (CPT) of the mosquito repellent composition.
[00124] The present invention provides a mosquito repellent composition, which is easy to use and provides long term protection.
[00125] The present invention provides a mosquito repellent composition, which has pleasant fragrance, and can be used indoors as well as outdoors.
[00126] The present invention provides a mosquito repellent composition, which alleviates the danger of diseases like Zika Fever, Dengue, Chikungunya, Malaria and Yellow fever.
[00127] The present invention provides a mosquito repellent composition, having cleansing and antimicrobial properties.
[00128] The present invention provides a mosquito repellent composition that provides antimicrobial efficacy of 99.99% against skin pathogenic bacteria like Staphylococcus aureus, Pseudomonas aeruginosa, Staphylococcus epidermidis and Candida albanicus.
CLAIMS:1. A mosquito repellent composition comprising natural surfactant, natural oils and plant extracts, skin conditioner, emulsifiers, preservative, pH adjuster and anti-microbial agent, wherein natural oils and plant extracts act as natural repellent and natural surfactant is Rhamnolipid.
2. The composition as claimed in claim 1, wherein natural oils and plant extracts are selected from Peppermint Oil, citronella oil, Lemon eucalyptus Oil, Tea Tree Oil, Calendula Oil, or combinations thereof.
3. The composition as claimed in claim 1, wherein skin conditioner can be selected from Thiamine Hydrochloride, Calendula Oil or combinations thereof.
4. The composition as claimed in claim 1, wherein Thiamine hydrochloride is a skin conditioner and mosquito repellent.
5. The composition as claimed in claim 1, wherein preservative can be selected from Nisin, dehydroacetic acid or combinations thereof.
6. The composition as claimed in claim 1, wherein Nisin is a natural preservative and anti-microbial agent.
7. The composition as claimed in claim 1, wherein emulsifier is selected from lecithin or sunflower lecithin.
8. The composition as claimed in claim 1, comprising :
a) Rhamnolipid;
b) Nisin;
c) Peppermint Oil;
d) Lemon eucalyptus Oil ;
e) Citronella Oil;
f) Tea tree Oil;
g) Calendula Oil ;
h) Thiamine hydrochloride;
i) Dehydroacetic acid;
j) Sunflower Lecithin;
k) Sodium Hydroxide; and
l) Distilled water.
9. The composition as claimed in claim 1, wherein natural surfactant is present in an amount ranging from about 0.01% to about 5% by weight of the composition, preferably in the amount ranging from about 0.05 % to about 3 % by weight of the composition.
10. The composition as claimed in claim 1, wherein natural oils are present in an amount ranging from about 5% to about 50% by weight of the composition, preferably in an amount ranging from about 25% to about 35% by weight of the composition.
11. The composition as claimed in claim 1, wherein Peppermint Oil is present in an amount ranging from about 0.5% to about 10%, preferably in the amount ranging from about 1 % to about 5 % by weight of the composition.
12. The composition as claimed in claim 1, wherein lemon-eucalyptus oil is present in an amount ranging from about 1% to about 10%, preferably in the amount ranging from about 3 % to about 7 % by weight of the composition.
13. The composition as claimed in claim 1, wherein Citronella Oil is present in an amount ranging from about 1% to about 10%, preferably in the amount ranging from about 3 % to about 7 % by weight of the composition.
14. The composition as claimed in claim 1, wherein tea tree oil is present in an amount ranging from about 2% to about 20% preferably in the amount ranging from about 5 % to about 15 % by weight of the composition.
15. The composition as claimed in claim 1, wherein calendula oil is present in an amount ranging from about 5% to about 30% preferably in the amount ranging from about 10 % to about 20 % by weight of the composition.
16. The composition as claimed in claim 1, wherein preservative is present in an amount ranging from about 0.001% to about 1% by weight of the composition, preferably in an amount ranging from about 0.005% to about 0.5% by weight of the composition.
17. The composition as claimed in claim 1, wherein skin conditioner is present in an amount ranging from about 0.1% to about 20% by weight of the composition, preferably in an amount ranging from about 0.5 % to about 15% by weight of the composition.
18. The composition as claimed in claim 1, wherein emulsifier is present in an amount ranging from about 0.5% to about 10% by weight of the composition, preferably in an amount ranging from about 1% to about 5% by weight of the composition.
19. The composition as claimed in claim 1, wherein anti-microbial agents are present in an amount ranging from about 0.001% to about 25% by weight of the composition, preferably in an amount ranging from about 0.01% to about 20% by weight of the composition.
20. The composition as claimed in claim 1, comprising :
a) Rhamnolipids 0.1g;
b) Nisin 0.025g;
c) Peppermint Oil 2ml;
d) Lemon eucalyptus Oil 5ml;
e) Citronella Oil 5ml;
f) Tea tree Oil 10ml;
g) Calendula Oil 15ml;
h) Thiamine hydrochloride 0.1g;
i) Dehydroacetic acid 0.03g;
j) Sunflower Lecithin 4ml;
k) Sodium Hydroxide: Adjusted using 10M Stock solution; and
l) Distilled water q.s. to make 100mL.
21. The composition as claimed in claim 1, which provides long term protection from mosquitoes in the range of 4-10 hours.
22. The composition as claimed in claim 1, which can be formulated as a deodorizer, waterless body bath, cream, lotion or oil and can be easily applied on the skin.
23. The composition as claimed in claim 1, which99.99% effective against bacterial cultures Staphylococcus aureus, Pseudomonas aeruginosa, Staphylococcus epidermidis and fungal culture Candida albanicus.