FIELD OF INVENTION
[001] The present disclosure broadly relates to field of biopharmaceuticals and particularly refers to a composition exhibiting anti-biofilm activity against the opportunistic human pathogens. The present disclosure also discloses a process of preparing the composition.
BACKGROUND OF INVENTION
[002] A biofilm is formed by the accumulation of microorganisms embedded in a polysaccharide matrix that is adhered to a biological or a non-biotic surface. Diverse micro-organisms such as fungi, bacteria, protozoa, and associated bacteriophage viruses can be found in these biofilms. These micro-organisms are planktonic, i.e., they move freely around in the water and other liquid media. However, there are many pathogenic and harmful microorganisms that get attached to a surface where they form biofilms.
[003] Bacteria in a biofilm can also survive harsh conditions and withstand the host's immune system. Especially the bacteria belonging to the genus Staphylococcus colonizes the human epithelium and turns pathogenic. In particular, the biofilm-forming ability of Methicillin-Resistant Staphylococcus aureus (MRSA) has made them very stubborn and strong. This commensal bacterium that turns pathogenic against humans becomes ineradicable by gaining antibiotic resistance. Microbes living in biofilms show an elevated pattern of adaptive resistance to antibiotics and other disinfectants compared to their planktonic compartments. Adaptive antibiotic resistance acts as an obstacle when treating biofilm-associated acute and chronic infections. Many currently available antibiotics are reported to have toxic side effects. Particularly, antibiotics disrupt gut microbiome and kill beneficial bacteria. Therefore, targeting biofilm formation using anti-biofilm has become an alternative strategy to the antibiotics. [004] Anti-biofilm agents disrupt the adherence of bacterium to a surface instead of killing the bacterial cells, thereby excluding the possibility of the emergence of antibiotic resistance. Various anti-biofilm agents are reported in the literature to inhibit the biofilm formation activity of bacteria. For instance, JP2016517396A

document discloses an anti-biofilm composition comprising a group of compounds such as disodium EDTA, sodium citrate, zinc chloride for use against bacteria, and bacterial biofilms associated with one or more of infections in wounds, cystic fibrosis infections, mastitis, otitis, food-borne diseases, and hospital-related infections. EP2296476A1 document discloses a method for inhibiting biofilm formation by bacterial cells on a solid surface comprising the step of contacting the surface with organic compounds, particularly thiazolidinedione, hydantoin, rhodamine compounds.
[005] While many improvements have been made to the anti-biofilm agents, there is still a dire need to provide anti-biofilm agents that are safe, effective in inhibiting the biofilm formation by Staphylococcus bacteria without any risk of emergence of resistance to the drugs.
SUMMARY OF THE INVENTION
[006] In an aspect of the present disclosure, there is provided a composition comprising: a) dodecanolide; and b) thymol, wherein dodecanolide has a weight percentage in a range of 0.0005-0.002% with respect to the composition, and thymol has a weight percentage in a range of 0.0001-0.001% with respect to the composition.
[007] In another aspect of the present disclosure, there is provided a process for preparing a composition comprising: a) dodecanolide; b) thymol; c) at least one water phase component and d) at least one oil phase component, wherein dodecanolide has a weight percentage in a range of 0.0005-0.002%) with respect to the composition, and thymol has a weight percentage in a range of 0.0001-0.001%) with respect to the composition, said process comprising: a) obtaining dodecanolide; b) obtaining thymol; and c) obtaining at least one water phase component or at least one oil phase component; d) contacting dodecanolide and thymol with the at least one water phase component or the at least one oil phase component to obtain the composition.

[008] In yet another aspect of the present disclosure, there is provided a process for preparing a composition comprising: a) dodecanolide; b) thymol; and c) a combination of at least one water phase and at least one oil phase component, wherein dodecanolide has a weight percentage in a range of 0.0005-0.002% with respect to the composition, and thymol has a weight percentage in a range of 0.0001-0.001% with respect to the composition, said process comprising: i) heating the at least one water phase component to a temperature in range of 65-75°C for a time period in a range of 25-35 minutes, to obtain a heated water phase component, and blending the heated water phase component at a speed in a range of 200-400 rpm for a time period in a range of 10-20 minutes to obtain a first mixture; ii) heating the at least one oil phase component at a temperature in a range of 60-80°C for a time period in a range of 25-35 minutes to obtain, to obtain a heated oil phase component, and blending the heated oil phase component at a speed of 300-500 rpm for a time period in a range of 20-40 minutes to obtain a second mixture; iii) contacting the first mixture and the second mixture at a speed in a range of 400-600 rpm for a time period in range of 2-40 minutes to obtain a third mixture; and iv) contacting dodecanolide, thymol, the at least one solvent and the third mixture at a speed in a range of 400-600 rpm for a time period in a range of 40-50 minutes at a temperature in a range of 35-40°C, to obtain the composition. [009] These and other features, aspects, and advantages of the present subject matter will be better understood with reference to the following description and appended claims. This summary is provided to introduce a selection of concepts in a simplified form. This summary is not intended to identify key features or essential features of the claimed subject matter, nor is it intended to be used to limit the scope of the claimed subject matter.
BRIEF DESCRIPTION OF ACCOMPANYING DRAWINGS
[0010] The following drawings form a part of the present specification and are included to further illustrate aspects of the present disclosure. The disclosure may be better understood by reference to the drawings in combination with the detailed description of the specific embodiments presented herein.

[0011] Figure 1 depicts the percentage inhibition of biofilm formed by MRSA in the
presence of dodecanolide at different concentrations, in accordance with an
embodiment of the present disclosure.
[0012] Figure 2 depicts the percentage inhibition of biofilm formed by MRSA in the
presence of thymol at different concentrations, in accordance with an embodiment of
the present disclosure.
[0013] Figure 3 depicts the various combinations of dodecanolide and thymol used
at different concentrations, in accordance with an embodiment of the present
disclosure.
[0014] Figure 4 depicts the representative images of Petri plates of Colony Forming
Unit (CFU) assay for the assessment of non-antibacterial activity against MRSA, in
accordance with an embodiment of the present disclosure.
[0015] Figure 5 depicts the assessment of the growth of MRSA upon treatment with
the combination of dodecanolide and thymol, in accordance with an embodiment of
the present disclosure.
DETAILED DESCRIPTION OF THE INVENTION
[0016] Those skilled in the art will be aware that the present disclosure is subject to variations and modifications other than those specifically described. It is to be understood that the present disclosure includes all such variations and modifications. The disclosure also includes all such steps, features, compositions, and compounds referred to or indicated in this specification, individually or collectively, and any and all combinations of any or more of such steps or features. Definitions
[0017] For convenience, before further description of the present disclosure, certain terms employed in the specification, and examples are delineated here. These definitions should be read in the light of the remainder of the disclosure and understood as by a person of skill in the art. The terms used herein have the meanings recognized and known to those of skill in the art, however, for

convenience and completeness, particular terms and their meanings are set forth
below.
[0018] The articles "a", "an" and "the" are used to refer to one or to more than one
(i.e., to at least one) of the grammatical object of the article.
[0019] The terms "comprise" and "comprising" are used in the inclusive, open
sense, meaning that additional elements may be included. It is not intended to be
construed as "consists of only".
[0020] Throughout this specification, unless the context requires otherwise the
word "comprise", and variations such as "comprises" and "comprising", will be
understood to imply the inclusion of a stated element or step or group of element or
steps but not the exclusion of any other element or step or group of element or steps.
[0021] The term "including" is used to mean "including but not limited to".
"Including" and "including but not limited to" are used interchangeably.
[0022] Phytochemicals (from Greek phyto, meaning "plant") are chemicals
produced by plants through primary or secondary metabolism. They generally have
biological activity in the plant host and play a role in plant growth or defense against
competitors, pathogens, or predators.
[0023] The term "biofilm" is used to describe the aggregation of microorganisms
in which cells adhere to each other to a surface (living/nonliving). The biofilm
formation confers numerous advantages to the pathogens, including an increase in
drug resistance and providing high tolerance to environmental stresses such as
antimicrobials and host immune responses.
[0024] As used herein, the water phase components are defined as components that
dissolve in water. Similarly, oil phase components are defined as components that
dissolve in oil. And in the present disclosure, the term water-phase components and
oil-phase components are added to the active ingredients, forming the composition
as disclosed in the present disclosure. Particularly,
[0025] For the purpose of the present disclosure, the term "preservatives",
"humectants", "chelants", "feel enhancer", "neutralizes", "emollients",
"structurants" refers to the generally known components used in the art.

[0026] The Biofilm Inhibitory Concentration (BIC) is defined as the lower concentration of antibiotics/ antimicrobials showing 50% and 90% reduction of biofilm binding. The percentage of biofilm inhibition is calculated by the following formula:

[0027] For the purposes of the present document, the term "synergism" is used to describe the interaction or cooperation of two or more organizations, substances, or other agents to produce a combined effect greater than the sum of their separate effects. The synergistic effect shown by the combination of two phytochemicals at various concentrations is studied by the checkerboard assay through Fractional Inhibitory Concentration Index (FIC). Therefore, FIC is used to estimate the interaction between two or more phytochemicals used in combination.
i) If the FIC index is <_0.5, then the combined effect shown by the two or
more phytochemicals used in combination is denoted as synergistic.
ii) If the FIC index is between 0.5 to 4, then no interaction is shown by two or more phytochemicals used in combination.
iii) If the FIC index is >4, then the combined effect shown by the two or more phytochemicals used in combination is denoted as antagonistic. [0028] Ratios, concentrations, amounts, and other numerical data may be presented herein in a range format. It is to be understood that such range format is used merely for convenience and brevity and should be interpreted flexibly to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited.
[0029] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the disclosure, the preferred methods, and materials are now described. All publications mentioned herein are incorporated herein by reference.

[0030] The present disclosure is not to be limited in scope by the specific embodiments described herein, which are intended for the purposes of exemplification only. Functionally-equivalent products, compositions, and methods are clearly within the scope of the disclosure, as described herein. [0031] As discussed in the background of the present disclosure, the current measures against the genus Staphylococcus bacterial biofilm-associated infections employ antibiotics. The antibiotics are inefficient at their effective concentration due to the emergence of antibiotic resistance and also creates an imbalance in the human microbiota. The majority of currently available antibiotics are reported to have toxic side effects. Therefore, targeting biofilm formation by different anti-biofilm agents has become an alternative strategy to antibiotics. Although many chemical compounds have been reported as anti-biofilm agents, however, their use for exhibiting anti-biofilm activity of bacteria may have many issues with cytotoxicity, bio-availability, and other side effects. Therefore, it has been aimed to develop an anti-biofilm agent against the microbes employing phytochemicals which are generally recognized as safe. Plenty of compounds with anti-biofilm activity have been identified from various natural resources so far. But still, many of the identified compounds have failed to cross the clinical trials because of the requirement of high concentration and toxic side effects.
[0032] To address the problems as mentioned above, the present disclosure provides a composition based on the natural compounds.
[0033] In the present invention, a synergistic combination of phytochemicals as anti-biofilm agent(s) is developed, since combinatorial therapy has the benefits of reduced toxicity, improved efficacy, and impedes the pathogen in developing resistance against two drug candidates concurrently. Moreover, the current disclosure is also aimed to potentiate the efficacy of currently available antibiotics by combining with anti-biofilm agents and the context of drug resistance was also vitally considered. Therefore, the current invention focuses on the development of cost-effective, non-toxic, and potential anti-biofilm formulation without developing antibiotic resistance.

[0034] The present disclosure pertains to the synergistic combination achieved through the use of phytochemicals comprising of dodecanolide and thymol, as mentioned herein. The phytochemicals are generally considered as GRAS (Generally recognized as safe) compounds The synergistic combination of dodecanolide and thymol provides inhibitory activity against biofilm formation in the genus of Staphylococcus. The present disclosure discloses a composition comprising phytochemicals such as dodecanolide and thymol. The phytochemicals when present in specific weight ranges as disclosed in the present disclosure provides a synergistic anti-biofilm effect against the infection caused by the Staphylococcus genus. Besides, the synergistic combination of dodecanolide and thymol exhibits anti-biofilm efficacy at lower concentrations with a decreased probability of developing drug resistance. It has also been established that the desirable synergistic effect is not exhibited if the composition is made of individual phytochemicals outside a specific weight range. The composition can be used as an anti-biofilm formulation for effectively inhibiting the biofilm formation by Methicillin-Resistant S. aureus (MRS A). One of the most important features of the composition of the present disclosure is that the synergistic combination of dodecanolide and thymol identified synergistic combinations do not affect the growth of MRSA and hence the possibility of drug resistance and toxicity is very meager.
[0035] In an embodiment of the present disclosure, there is provided a composition comprising: a) dodecanolide; and b) thymol, wherein dodecanolide has a weight percentage in a range of 0.0005-0.002% with respect to the composition, and thymol has a weight percentage in a range of 0.0001-0.001% with respect to the composition.
[0036] In an embodiment of the present disclosure, there is provided a composition comprising: a) dodecanolide; and b) thymol, wherein dodecanolide has a weight percentage in a range of 0.0005-0.002%) with respect to the composition. In another embodiment of the present disclosure, dodecanolide has a weight percentage in a range of 0.0008-0.0018% with respect to the composition. In yet another embodiment of the present disclosure, dodecanolide has a weight percentage in a

range of 0.0010-0.0016% with respect to the composition. In one another embodiment of the present disclosure, dodecanolide has a weight percentage in a range of 0.0012-0.0015% with respect to the composition.
[0037] In an embodiment of the present disclosure, there is provided a composition comprising: a) dodecanolide; and b) thymol, wherein thymol has a weight percentage in a range of 0.0001-0.001% with respect to the composition. In another embodiment of the present disclosure, thymol has a weight percentage in a range of 0.0003-0.0009% with respect to the composition In yet another embodiment of the present disclosure, thymol has a weight percentage in a range of 0.0005-0.0008%) with respect to the composition. In one another embodiment of the present disclosure, thymol has a weight percentage in a range of 0.0006-0.0007%) with respect to the composition.
[0038] In an embodiment of the present disclosure, there is provided a composition comprising: a) dodecanolide; b) thymol; and c) at least one water phase component, wherein dodecanolide has a weight percentage in a range of 0.0005-0.002%) with respect to the composition, thymol has a weight percentage in a range of 0.0001-0.001%) with respect to the composition and at least one water phase component has weight percentage in the range of 0.025-11.0% with respect to the composition. [0039] In an embodiment of the present disclosure, there is provided a composition comprising: a) dodecanolide; b) thymol; and c) at least one oil phase component, wherein dodecanolide has a weight percentage in a range of 0.0005-0.002%) with respect to the composition, thymol has a weight percentage in a range of 0.0001-0.001%) with respect to the composition and at least one oil phase component has weight percentage in the range of 0.07 -53.0% with respect to the composition. [0040] In an embodiment of the present disclosure, there is provided a composition comprising: a) dodecanolide; b) thymol and c) a combination of at least one water phase component and at least one oil phase component, wherein dodecanolide has a weight percentage in a range of 0.0005-0.002%) with respect to the composition, and thymol has a weight percentage in a range of 0.0001-0.001%) with respect to the composition.

[0041] In an embodiment of the present disclosure, there is provided a composition comprising: a) dodecanolide; b) thymol; and c) at least one water phase component, wherein dodecanolide has a weight percentage in a range of 0.0005-0.002% with respect to the composition, and thymol has a weight percentage in a range of 0.0001-0.001% with respect to the composition, wherein the at least one water phase component is selected from a group consisting of preservatives, humectants, chelants, and combinations thereof. In another embodiment of the present disclosure, the preservatives are selected from a group consisting of paraben, gluconolactone, sodium benzoate, chlorobutanol, benzyl alcohol, imidazolidinyl urea, benzalkonium chloride, alpha tocopherol, sodium sulphite, methyl-dibromo glutaronitrile, tea tree essential oil, and combinations thereof, the humectants are selected from a group consisting of aloe vera powder, arnica extract, algae extract, hydrolyzed baobab protein, coenzyme qlO, hydrolyzed collagen protein, colloidal oatmeal, glycerin, caprylyl glycol, ceramide complex, lacto-ceramide, matrixyl s-6, propylene glycol, rhubarb root extract, hyaluronic acid, hydrolyzed jojoba protein, hydrolyzed keratin protein, sodium PCA, and combinations thereof, the chelants are selected from a group consisting of diethylenetriaminepentaacetic acid, tetrasodium ethylenediaminetetraacetate, disodium ethylenediaminetetraacetate, triglycollamic acid, ethane 1-hydroxy-1, 1-diphosphonate, polyphosphates, citric acid, and combinations thereof.
[0042] In an embodiment of the present disclosure, there is provided a composition comprising: a) dodecanolide; b) thymol; and c) at least one oil phase component, wherein dodecanolide has a weight percentage in a range of 0.0005 - 0.002 % with respect to the composition, and thymol has a weight percentage in a range of 0.0001 - 0.001 % with respect to the composition, and wherein the at least one oil phase component is selected from a group consisting of feel enhancers, neutralizers, emollients, structurants, and combinations thereof. In another embodiment of the present disclosure, the feel enhancers are selected from a group consisting of isopropyl myristate, bis-di glyceryl polyacyladipate-1, butylene glycol dicaprylate/dicaprate, cetearyl isononanoate, decyl oleate, glyceryl oleate, glyceryl ricinoleate, oleyl erucate, olus oil, alkyl citrate and stearate esters, waxes, essential

oils, and combinations thereof, the neutralizers are selected from a group consisting of borax, sodium hydroxide, sodium carbonate and alkali extract of coconut husk, triethanol amine, and combinations thereof, the emollients are selected from a group consisting of amodimethicone, C12-15 alkyl benzoate, cetyl palmitate, isopropyl myristate, jojoba gel, mineral oil, natural gel-wax, octyldodecanol, amodimethicone, cyclodimethicone, cyclomethicone, dimethicone 500, dimethicone satin, isodimethicone copolymer, PEG8, polyisobutene 250, squalane, almond oil, castor oil, grapeseed oil, red raspberry seed oil, cherry kernel oil, and combinations thereof, and the structurants are selected from a group consisting of rice bran oil, corn oil, sunflower oil, starch, alginate, carnauba wax, bees wax, berry wax, fruit wax, silicone based polymers, acrylate copolymer, cellulose based polymers, and combinations thereof.
[0043] In an embodiment of the present disclosure, there is provided a composition comprising: a) dodecanolide; b) thymol; and c) at least one water phase component, wherein dodecanolide has a weight percentage in a range of 0.0005-0.002% with respect to the composition, and thymol has a weight percentage in a range of 0.0001-0.001% with respect to the composition, wherein the at least one water phase component is selected from a group consisting of preservatives having a weight percentage in a range of 0.005-0.5%) with respect to the composition, humectants a weight percentage in a range of 0.01-10%) with respect to the composition, chelants having a weight percentage in a range of 0.01-0.5%) with respect to the composition.
[0044] In an embodiment of the present disclosure, there is provided a composition comprising: a) dodecanolide; b) thymol; and c) at least one oil phase components, wherein dodecanolide has a weight percentage in a range of 0.0005-0.002%) with respect to the composition, and thymol has a weight percentage in a range of 0.0001-0.001%) with respect to the composition, wherein the at least one oil phase components is selected from a group consisting of feel enhancers having a weight percentage in range of 0.01-25%> with respect to the composition, neutralizers having a weight percentage in a range of 0.0004-0.4%> with respect to the composition, emollients having a weight percentage in a range of 0.05-20%> with

respect to the composition, structurants having a weight percentage in a range of 0.01-7% with respect to the composition.
[0045] In an embodiment of the present disclosure, there is provided a composition comprising: a) dodecanolide; b) thymol; and c) a combination of at least one water phase and at least one oil phase component, wherein dodecanolide has a weight percentage in a range of 0.0005 - 0.002 % with respect to the composition, thymol has a weight percentage in a range of 0.0001 - 0.001 % with respect to the composition and combination of at least one water phase and at least one oil phase component has weight percentage in the range of 0.07-64% with respect to the composition.
[0046] In an embodiment of the present disclosure, there is provided a composition comprising: a) dodecanolide; b) thymol; and c) a combination of at least one water phase and at least oil phase component, wherein dodecanolide has a weight percentage in a range of 0.0005 - 0.002 % with respect to the composition, and thymol has a weight percentage in a range of 0.0001 - 0.001 % with respect to the composition, and wherein the at least one water phase is selected from a group consisting of preservatives having a weight percentage in a range of 0.005-0.5%) with respect to the composition, humectants a weight percentage in a range of 0.01-10%o with respect to the composition, chelants having a weight percentage in a range of 0.01-0.5%) with respect to the composition, and the at least one oil phase component is selected from a group consisting of feel enhancers having a weight percentage in range of 0.01-25%) with respect to the composition, neutralizers having a weight percentage in a range of 0.0004-0.4% with respect to the composition, emollients having a weight percentage in a range of 0.05-20%> with respect to the composition, structurants having a weight percentage in a range of 0.01-7%) with respect to the composition.
[0047] In an embodiment of the present disclosure, there is provided a composition comprising: a) dodecanolide; and b) thymol, wherein dodecanolide has a weight percentage in a range of 0.0005-0.002%) with respect to the composition, and thymol has a weight percentage in a range of 0.0001-0.001%> with respect to the composition, wherein the composition exhibits inhibitory activity against biofilm

formation of at least one microorganism selected from a group consisting of bacteria, fungi, protozoans. In another embodiment of the present disclosure, the at least one microorganism is bacteria.
[0048] In an embodiment of the present disclosure, there is provided a composition comprising: a) dodecanolide; b) thymol; and c) at least one water phase component, wherein dodecanolide has a weight percentage in a range of 0.0005 - 0.002 % with respect to the composition, and thymol has a weight percentage in a range of 0.0001
- 0.001 % with respect to the composition, and wherein the least one water phase
component is selected from a group consisting of preservatives, humectants,
chelants, and combinations thereof, and wherein the composition exhibits
inhibitory activity against biofilm formation of at least one microorganism selected
from a group consisting of bacteria, fungi, protozoans. In another embodiment of
the present disclosure, the at least one microorganism is bacteria.
[0049] In an embodiment of the present disclosure, there is provided a composition comprising: a) dodecanolide; b) thymol; and c) at least one oil phase component, wherein dodecanolide has a weight percentage in a range of 0.0005 - 0.002 % with respect to the composition, and thymol has a weight percentage in a range of 0.0001
- 0.001 % with respect to the composition, wherein the at least one oil phase
component is selected from a group consisting of feel enhancers, neutralizers,
emollients, structurants, and combinations thereof, and wherein the composition
exhibits inhibitory activity against biofilm formation of at least one microorganism
selected from a group consisting of bacteria, fungi, protozoans. In another
embodiment of the present disclosure, the at least one microorganism is bacteria.
[0050] In an embodiment of the present disclosure, there is provided a composition
comprising: a) dodecanolide; b) thymol; and c) a combination of at least one water
phase and at least oil phase component, wherein dodecanolide has a weight
percentage in a range of 0.0005 - 0.002 % with respect to the composition, and
thymol has a weight percentage in a range of 0.0001 - 0.001 % with respect to the
composition, and wherein the at least one water phase is selected from a group
consisting of preservatives, humectants, chelants, and combinations thereof, and the
at least one oil phase component is selected from a group consisting of feel

enhancers, neutralizers, emollients, structurants, and combinations thereof, and wherein the composition exhibits inhibitory activity against biofilm formation of at least one microorganism selected from a group consisting of bacteria, fungi, protozoans. In another embodiment of the present disclosure, the at least one microorganism is bacteria.
[0051] In an embodiment of the present disclosure, there is provided a composition comprising: a) dodecanolide; and b) thymol, wherein dodecanolide has a weight percentage in a range of 0.0005-0.002% with respect to the composition, and thymol has a weight percentage in a range of 0.0001-0.001% with respect to the composition, wherein the at least one microorganism is bacteria. [0052] In an embodiment of the present disclosure, there is provided a composition comprising: a) dodecanolide; and b) thymol, wherein dodecanolide has a weight percentage in a range of 0.0005-0.002%) with respect to the composition, and thymol has a weight percentage in a range of 0.0001-0.001%) with respect to the composition, wherein said composition exhibits inhibitory activity against biofilm formation by bacteria is selected from a group consisting of Staphylococcus aureus, Methicillin-resistant Staphylococcus aureus (MRSA), Methicillin-susceptible Staphylococcus aureus (MSSA) and combinations thereof.
[0053] In an embodiment of the present disclosure, there is provided a composition comprising: a) dodecanolide; and b) thymol, wherein dodecanolide has a weight percentage in a range of 0.0005-0.002%) with respect to the composition, and thymol has a weight percentage in a range of 0.0001-0.001%> with respect to the composition, wherein the composition has a shelf life for a period in a range of 12 months to 24 months. In another embodiment of the present disclosure, the composition has a shelf life for a period in a range of 14-22 months. In yet another embodiment of the present disclosure, the composition has a shelf life for a period in a range of 16-20 months.
[0054] In an embodiment of the present disclosure, there is provided a process for preparing a composition comprising: a) dodecanolide; and b) thymol, wherein dodecanolide has a weight percentage in a range of 0.0005-0.002 % with respect to the composition, and thymol has a weight percentage in a range of 0.0001-0.001%>

with respect to the composition, said process comprising: a) obtaining dodecanolide; b) obtaining thymol; c) obtaining at least one water phase component or at least one oil phase component; and d) contacting dodecanolide and thymol with the at least one water phase component or the at least one oil phase component to obtain the composition.
[0055] In an embodiment of the present disclosure, there is provided a process for preparing a composition comprising: a) dodecanolide; b) thymol; and c) at least one water phase component or the at least one oil phase component, wherein dodecanolide has a weight percentage in a range of 0.0005-0.002 % with respect to the composition, and thymol has a weight percentage in a range of 0.0001-0.002% with respect to the composition, said process comprising: a) obtaining dodecanolide; b) obtaining thymol; c) obtaining at least one water phase component or at least one oil phase component; and d) contacting dodecanolide and thymol with the at least one water phase component or the at least one oil phase component to obtain the composition.
[0056] In an embodiment of the present disclosure, there is provided a process for preparing a composition comprising: a) dodecanolide; b) thymol; and c) at least one water phase component or at least one oil phase component, wherein dodecanolide has a weight percentage in a range of 0.0005-0.002 % with respect to the composition, and thymol has a weight percentage in a range of 0.0001-0.001% with respect to the composition, said process comprising: a) obtaining dodecanolide; b) obtaining thymol; c) obtaining at least one water phase component or at least one oil phase component; and d) contacting dodecanolide and thymol with the at least one water phase component or the at least one oil phase component to obtain the composition., wherein the process further comprises at least one solvent having a weight percentage in a range of 10-98% with respect to the composition. [0057] In an embodiment of the present disclosure, there is provided a process for preparing a composition comprising: a) dodecanolide; b) thymol; and c) at least one water phase component or at least one oil phase component, wherein dodecanolide has a weight percentage in a range of 0.0005-0.002 % with respect to the composition, and thymol has a weight percentage in a range of 0.0001-0.001% with

respect to the composition, said process comprising: a) obtaining dodecanolide; b) obtaining thymol; c) obtaining at least one water phase component or at least one oil phase component; and d) contacting dodecanolide and thymol with the at least one water phase component or the at least one oil phase component to obtain the composition, wherein the process further comprises at least one solvent selected from a group consisting of water, methanol, ethanol, dimethyl sulfoxide (DMSO), and combinations thereof, having a weight percentage in a range of 10-98% with respect to the composition. In another embodiment of the present disclosure, the at least one solvent is water.
[0058] In an embodiment of the present disclosure, there is provided a process for preparing a composition comprising: a) dodecanolide; and b) thymol, wherein dodecanolide has a weight percentage in a range of 0.0005-0.002 % with respect to the composition, and thymol has a weight percentage in a range of 0.0001-0.001% with respect to the composition, said process comprising: a) heating the at least one water phase component to a temperature in a range of 65-75°C for a time period in a range of 25-35 minutes, to obtain a heated water phase component, and blending the heated water phase component at a speed in a range of 200-400 rpm for a time period in a range of 10-20 minutes to obtain a first mixture; b) heating the at least one oil phase component at a temperature in a range of 60-80°C for a time period in a range of 25-35 minutes to obtain a heated oil phase component, and blending the heated oil phase component at a speed of 300-500 rpm for a time period in a range of 20-40 minutes to obtain a second mixture; c) contacting the first mixture and the second mixture at a speed in a range of 400-600 rpm for a time period in range of 2-40 minutes to obtain a third mixture; and d) contacting dodecanolide, thymol, the at least one solvent and the third mixture at a speed in a range of 400-600 rpm for a time period in a range of 40-50 minutes at a temperature in a range of 35-40°C, to obtain the composition.
[0059] In an embodiment of the present disclosure, there is provided a formulation comprising the composition as described herein, wherein the formulation can be used in a form of an anti-biofilm cream, ointment, lotion, paste, gel or the like.

[0060] Although the subject matter has been described in considerable detail with reference to certain examples and implementations thereof, other implementations are possible. EXAMPLES
[0061] The disclosure will now be illustrated with working examples, which is intended to illustrate the working of disclosure and not intended to take restrictively to imply any limitations on the scope of the present disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this disclosure belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice of the disclosed methods and compositions, the exemplary methods, devices and materials are described herein. It is to be understood that this disclosure is not limited to particular methods, and experimental conditions described, as such methods and conditions may apply. [0062] The working examples as depicted in the forthcoming sections highlight the combinatorial effect of different phytochemicals used at different concentrations on Methicillin Resistant Staphylococcus aureus (MRSA). The composition as disclosed in the present disclosure comprised: a) dodecanolide; and b) thymol, wherein dodecanolide has a weight percentage in a range of 0.0005-0.002 % with respect to the composition, and thymol has a weight percentage in a range of 0.0001-0.001% with respect to the composition. The synergism between dodecanolide and thymol is achieved when the combinatorial effect is greater than the sum of individual phytochemical effect against the biofilm formation by MRSA. The absence of any of the phytochemical specified above or replacement of the same with any other phytochemical substantially diminishes the synergistic effect against the biofilm formation.
Materials and Methods
[0063] The phytochemicals such as dodecanolide and thymol were used for arriving at the composition of the present disclosure. Dodecanolide and thymol were procured from Sigma-Aldrich and Alfa-Aesar, respectively. The anti-biofilm

efficacy of the phytochemicals mentioned above was studied in the Methicillin-Resistant S. aureus (MRSA) species.
Evaluation of Biofilm Inhibitory Concentration (BIC) of dodecanolide and thymol against biofilm formed by Methicillin-Resistant S. aureus (MRSA): [0064] Different concentrations of dodecanolide and thymol were used against the species of MRSA. The effect of dodecanolide and thymol on biofilm formed by MRSA at different concentrations was evaluated to determine the Biofilm Inhibitory Concentration (BIC) of the respective drug. The BIC of dodecanolide and thymol against MRSA biofilm was evaluated by broth dilution assay in 24-well microtitre plates (MTP). For this purpose, each well of MTP was loaded with 1 mL of Typtone soya broth containing 1% of sucrose (TSBS). 1% (v/v) of 12 h MRSA was allowed to form biofilm on 24-well Micro Titer Plate (MTP) treated with dodecanolide at different concentration in a range of 0 to 250 ug/mL and thymol at different concentrations in a range of 0 to 200 ug/mL. TSBS with MRSA culture alone was maintained as control. After incubation, the loosely bound planktonic cells were removed, and the biofilm attached to the MTP plate was washed twice with sterile saline and air-dried. Then, 0.4 % of crystal violet was added to each well and incubated for 5 min at room temperature, and was subsequently washed and air-dried. After that, the biofilms were destained using 10 % glacial acetic acid. The absorbance of the control and treated wells was measured at 570 nm. The percentage of biofilm inhibition was calculated using the formula:

[0065] Checkerboard assay - Checkerboard assay is an approach of studying the effect of the potency of the combination of phytochemicals in comparison to the effect of the potency of individual phytochemicals. The effect of the potency of the combination of phytochemicals in comparison to the effect of the potency of individual phytochemicals is determined by the Fractional Inhibitory Concentration (FIC) index. The phytochemical combinatorial effect is defined by the FIC index value.

• If FIC index is <0.5, then the phytochemical combinatorial effect is denoted as a synergistic effect.
• If FIC index is between 0.5 to 4.0, then the phytochemical combinatorial effect is indifferent.
• If FIC index is >4.0, then the phytochemical combinatorial effect is denoted as an antagonistic effect.
[0066] In the present disclosure, the combinatorial effect of dodecanolide and thymol was evaluated by using Checkerboard assay. Dodecanolide and thymol were used in a combination to inhibit the biofilm formed by MRSA. To study the effectiveness of the combined effect of dodecanolide and thymol against biofilm formed by MRSA, FIC index value was determined. The FIC index was determined as prescribed by ODDs, 2003 as follows:

Example 1
a) Effect of dodecanolide on MRSA
[0067] The effect of dodecanolide against the biofilm formed by MRSA was studied by using dodecanolide at different concentrations ranging from 0 to 250 ug/mL. The biofilm structure formed by MRSA was inhibited in a concentration-dependent manner without inhibiting the growth of the MRSA species. Figure 1 illustrates the percentage inhibition of biofilm formed by MRSA in the presence of

dodecanolide at different concentrations. Referring to Figure 1, the maximum
inhibition of MRSA biofilm was observed by using dodecanolide at a concentration
of 225 ug/mL.
b) Effect of thymol on MRSA
[0068] The effect of thymol against the biofilm formed by MRSA was studied by
using thymol at varying concentrations ranging from 0 to 200 ug/mL. Figure 2
illustrates the percentage inhibition of biofilm formed by MRSA in the presence of
thymol at different concentrations. Referring to Figure 2, thymol at 100 ug/mL
showed significant inhibition of biofilm formed by MRSA.
[0069] Hence, it can be inferred from Figure 1 and 2 that BIC of dodecanolide and
thymol was determined at 225 ug/mL and 100 ug/mL, respectively.
Example 2
Effect of composition of the present disclosure on MRSA biofilm formation [0070] Various combinations of dodecanolide and thymol were used against biofilm formation by MRSA. The combinatorial effect of dodecanolide and thymol against biofilm produced by MRSA was evaluated by checkerboard assay (as explained above in the method section). Figure 3 illustrates that various combinations of dodecanolide and thymol were evaluated at different concentrations to arrive at the composition of the present disclosure. This layout of the combination of the aforementioned phytochemicals was followed for MRSA biofilm inhibition study. It can be deduced from the checkerboard assay that if the combinatorial effect of two phytochemicals is found to be greater than the sum of individual effect, then there is a synergistic effect between the two phytochemicals. However, the inverse of the synergistic effect is antagonistic effect, and the intermediate between two effects is indifferent. a) Evaluation of the composition on the biofilm formation by MRSA:
[0071] Combinations of components such as dodecanolide and thymol at different concentrations were utilized to assess their anti-biofilm activity against MRSA, and their activity was evaluated by using checkerboard assay. In the combinations of dodecanolide and thymol, dodecanolide was varied from 14.0625 to 225 ugm/mL

and thymol was varied from 6.25 to 100 ug/mL. Table 1 depicts the FIC indexes of dodecanolide and thymol at varied concentration against MRSA biofilm formation. As depicted in the Table 1, the sign '*' indicated the presence and '-' indicated the absence of anti-biofilm activity in individual and combination of phytochemicals, respectively. Table 1

[0072] From Table 1, it can be inferred that most of the combinations of dodecanolide and thymol showed anti-biofilm activity, however, only a combination of dodecanolide and thymol at and 14.0625+6.25 ug/mL exhibited the maximum synergistic effect on inhibiting the biofilm formation as FIC index was <0.5, i.e., 0.125.
[0073] It can also be observed that the combination of dodecanolide and thymol at 56.25+25 ug/mL showed a substantial synergistic effect on inhibiting the biofilm formation as FIC index was 0.5. Further, it was observed that the combination of dodecanolide and thymol used at various concentrations such as 225, 112.5 and 28.125 ug/mL of dodecanolide and 100, 50, 25 and 12.5 ug/mL of thymol did not show the synergistic effect in inhibiting the biofilm formed by MRSA, with FIC indices greater than 0.5. It can also be inferred that the weight percentages of the

dodecanolide and thymol used for the studies were falling outside the weight percentage ranges as disclosed in the present disclosure.
[0074] Henceforth, it can be concluded that the composition having a combination of dodecanolide and thymol at a low concentration of 14.0625+6.25 ug/mL showed the synergistic effect against the biofilm formation in MRSA. Therefore, a combination of dodecanolide and thymol 14.0625+6.25 ug/mL is considered as the best working example, whereas, combination of dodecanolide and thymol used at various concentrations such as 225, 112.5 and 28.125 ug/mL of dodecanolide and 100, 50, 25 and 12.5 ug/mL of thymol were considered as the non-working examples.
b) The non-antibacterial activity of compositions against MRSA
[0075] The non-antibacterial activity of synergistic combinations of dodecanolide and thymol was confirmed by using colony-forming unit (CFU) assay. The composition having a combination of dodecanolide and thymol at 14.0625+6.25 ug/mL was added separately to the tube containing 1 mL of TSBS. Tubes without any treatment and tubes containing TSBS alone were deliberated as control and blank, respectively. All tubes were inoculated with 1% v/v of an overnight culture of MRSA and incubated at 37°C for 24 h in a shaker incubator at a speed of 160 rpm. After incubation, the absorbance of the control and treated cells was measured at 600 nm, and then cells were harvested by centrifugation (8000 rpm for 10 min) and subsequently suspended in 1 mL of Phosphate Buffered Saline (PBS). The cell suspensions were serially diluted, and 100 uL of 10"9 dilution of each sample was spread over the TSBS agar plates. TSBS agar plate spread with 100 uL of PBS was considered as blank. The plates were incubated overnight at 37°C for 24 h. Then, the number of colonies in each plate were counted manually and the graph was plotted. The number of colonies in treated samples were compared with that of control at an optical density of 600 nm.
[0076] Figure 4 depicts the representative images of Petri plates of CFU assay of the blank, control, and the composition having a combination of dodecanolide and

thymol at 14.0625+6.25 ug/mL against MRSA biofilm. Figure 5 depicts the assessment of the growth of MRSA upon treatment with the said composition of dodecanolide and thymol at 14.0625+6.25 ug/mL. As can be seen from Figure 4, the number of colonies was almost similar in both the control and treated Petri plates. Similarly, it can be observed from Figure 5 that the number of colonies in treated samples was comparable with that of control, and the optical density at 600nm was also unaffected upon treatment with said composition on MRSA biofilm. These results altogether substantiated the non-antibacterial nature of the synergistic combination of dodecanolide and thymol at 14.0625+6.25 ug mLused for arriving at the composition of the present disclosure. Hence, the composition is non-antibacterial. Example 3
(a) Process for preparing the composition of the present disclosure
[0077] The process for preparing the composition of the present disclosure comprised the following steps: (a) obtaining 0.0014% dodecanolide; (b) obtaining 0.00625% of thymol; (c) contacting dodecanolide and thymol to obtain the composition.
(b) Process for preparing the composition having a combination of water
phase and oil phase component of the present disclosure
[0078] In the present disclosure, the formulation was prepared by using the active ingredients as disclosed in the composition of the present disclosure. The formulation such as a topical cream or lotion comprising active ingredients along with water and oil phase components such as preservative, feel enhancer, neutralizer, humectant, emollient, structurant, chelant, solvent, and combinations thereof. Formulation, as disclosed in the present disclosure, can be used in the form of topical cream or lotion.
[0079] For the purpose of preparing the formulation, water phase and oil phase components were used.
[0080] Water phase components that were used in the present disclosure: 1 .Preservatives such as paraben, gluconolactone, sodium benzoate, chlorobutanol, benzyl alcohol, imidazolidinyl urea, benzalkonium chloride, alpha tocopherol,

sodium sulphite, methyl-dibromo glutaronitrile, Tea tree essential oil were used at the range of 0.005% to 0.5%.
2.Humectants such as aloe vera powder, arnica extract, algae extract, hydrolyzed baobab protein, coenzyme qlO, hydrolyzed collagen protein , colloidal oatmeal, glycerin, caprylyl glycol, ceramide complex, lacto-ceramide, matrixyl s-6, propylene glycol, Rhubarb root extract, hyaluronic acid, hydrolyzed jojoba protein, hydrolyzed keratin protein, sodium PCA were used at the range of 0.01% to 10%.
3.Chelants such as diethylenetriaminepentaacetic acid, tetrasodium
ethyl enediaminetetraacetate, disodium ethylenediaminetetraacetate,
triglycollamic acid, ethane 1-hydroxy-1, 1-diphosphonate, polyphosphates, citric acid were used at the range of 0.01% to 0.5%. [0081] Oil phase components that were used in the present disclosure:
1. Feel enhancers such as Isopropyl myristate, Bis-diglyceryl polyacyladipate-1, butylene glycol dicaprylate/dicaprate, cetearyl isononanoate, decyl oleate, glyceryl oleate, glyceryl ricinoleate, oleyl erucate, olus oil, alkyl citrate and stearate esters, waxes, essential oils were used at the range of 0.01% to 25%.
2. Neutralizers such as borax, sodium hydroxide, sodium carbonate, and alkali extract of coconut husk, triethanolamine were used at the range of 0.0004% to 0.4%.
3.Emollients such as amodimethicone, CI2-15 Alkyl benzoate, cetyl palmitate, isopropyl myristate, jojoba gel, mineral oil, natural gel-wax, octyldodecanol, amodimethicone, cyclodimethicone, cyclomethicone, dimethicone 500, dimethicone satin, isodimethicone copolymer, PEG8, polyisobutene 250, squalane, almond oil, castor oil, grapeseed oil, red raspberry seed oil, cherry kernel oil were used at the range of 0.05% to 20%.
4. Structurants such as rice bran oil, corn oil, sunflower oil, starch, alginate, carnauba wax, bees wax, berry wax, fruit wax, silicone based polymers, acrylate copolymer, cellulose based polymers were used at the range of 0.01 to 7%.

[0082] The solvent such as methanol, ethanol, or Dimethyl Sulfoxide (DMSO) and water were used in the range of 10% to 98%. Overall, the components used for preparing the formulation are shown in Table 2 below. Table 2

[0083] Table 2 depicts a formulation table comprising the composition of the present disclosure. According to the present disclosure, the formulation was prepared by using the active ingredients, i.e., synergistic combinations of dodecanolide (0.0014%) and thymol (0.00625%) along with other mentioned ingredients in the desired quantity and the working example for the formulation as depicted in table 2.
[0084] The formulation of the synergistic combination of dodecanolide and thymol as disclosed in the present disclosure was prepared by the following steps:
1. The water (aqueous) phase was prepared by dissolving the water phase components such as sodium benzoate, glycerin, and disodium EDTA in a required quantity of water.
2. The aqueous phase was kept in 70°C water bath for 30 minutes and stirred for 15 minutes at 300 rpm until all the components were completely dissolved to obtain the first mixture.
3. In another set, the oil phase components such as isopropyl myri state, triethanolamine, almond oil, and berry wax were taken in a required quantity and molten at 70°C for 30 minutes and stirred for 30 minutes at 400 rpm to obtain a better solubilized second mixture.
4. Then, the first mixture containing the aqueous phase was slowly added to the second mixture of oil phase with moderate agitation and was stirred for 30 minutes at 500 rpm, to obtain the third mixture
5. The active ingredients in the present disclosure, i.e., dodecanolide and thymol were dissolved in methanol at a stock concentration of 1 %, separately.
6. Dodecanolide and thymol combination at their working concentration of 0.0014%) and 0.00625%), respectively, were added to this mixture and stirred for 45 minutes at 500 rpm at 35-40°C.
7. The formulation was cooled to room temperature, and pH range of the formulation was measured to be 5-6.

8. Then, the formulation was stored at room temperature without direct exposure to sunlight with shelf life in the time period range of 12 to 26 months from the date of preparation.
[0085] Overall, the combination of the phytochemicals such as dodecanolide and thymol in the disclosed weight percentage ranges 0.0005-0.002 % and 0.0001-0.001% respectively were essential to have a desired synergistic effect against the biofilm formation of MRSA. Any deviation of the composition from their disclosed weight percentage ranges would not lead to the desired synergistic effect on inhibiting the biofilm activity of MRSA.
Advantages of the present disclosure
[0086] The present disclosure discloses a composition comprising dodecanolide and thymol, wherein dodecanolide has a weight percentage in a range of 0.0005-0.002% with respect to the composition, and thymol has a weight percentage in a range of 0.0001-0.001% with respect to the composition. The combination of the phytochemicals provides a synergistic effect against the biofilm formation in MRSA species, even by using these phytochemicals at a very low concentration. The efficacy of the phytochemicals is increased by the synergistic effect of using the combination of the composition as disclosed in the present disclosure. The combination of phytochemicals in present disclosure helps in overcoming the toxic side effects of antibiotics and antibiotic resistance. Further, their activity does not affect the viability of cells. The issue of further developing multidrug resistance is also addressed by the composition of the present disclosure, wherein the composition impedes the bacteria in developing resistance against dodecanolide and thymol concurrently. The composition of the present disclosure is also used for preparing the formulation. The formulation comprising the composition can be further used as a topical application such as anti-biofilm cream, ointment, lotion, paste, gel, etc. The present disclosure also discloses a convenient process for preparing the composition.

I/We Claim:
1. A composition comprising:
a. dodecanolide; and
b. thymol,
wherein dodecanolide has a weight percentage in a range of 0.0005-0.002% with respect to the total weight of the composition, and thymol has a weight percentage in a range of 0.0001-0.001% with respect to the total weight of the composition.
2. The composition as claimed in claim 1, wherein the composition includes at least one water phase component having a weight percentage in a range of 0.025-11.0% with respect to the total weight of the composition .
3. The composition as claimed in claim 1, wherein the composition includes at least one oil phase component having a weight percentage in a range of 0.07-53.0%) with respect to the total weight of the composition.
4. The composition as claimed in claim 1, wherein the composition includes a combination of at least one water phase and at least one oil phase component having a weight percentage in a range of 0.07-64.0%) with respect to the total weight of the composition.
5. The composition as claimed in claim 2 or 4, wherein the at least one water phase component is selected from a group consisting of preservatives, humectants, chelants, and combinations thereof.
6. The composition as claimed in claim 3 or 4, wherein the at least one oil phase component is selected from a group consisting of feel enhancers, neutralizers, emollients, structurants, and combinations thereof.
7. The composition as claimed in claim 5, wherein the preservatives has a weight percentage in a range of 0.005-0.5%) with respect to the composition, humectants has a weight percentage in a range of 0.01-10%) with respect to the composition, chelants has a weight percentage in a range of 0.01-0.5%) with respect to the composition.
8. The composition as claimed in claim 6, wherein the feel enhancers has a weight percentage in range of 0.01-25%) with respect to the composition,

neutralizers has a weight percentage in a range of 0.0004-0.4% with respect to the composition, emollients has a weight percentage in a range of 0.05-20% with respect to the composition, and structurants has a weight percentage in a range of 0.01-7% with respect to the composition.
9. The composition as claimed in any one of the claims 1 to 8, wherein the composition exhibits inhibitory activity against biofilm formation of at least one microorganism selected from a group consisting of bacteria, fungi, protozoans.
10. The composition as claimed in claim 9, wherein the at least one microorganism is bacteria.
11. The composition as claimed in claim 10, wherein the bacteria is selected from a group consisting of Staphylococcus aureus, Methicillin-resistant Staphylococcus aureus (MRSA), Methicillin-susceptible Staphylococcus aureus (MSSA) and combinations thereof.
12. The composition as claimed in any one of the claims 1 to 4, wherein the composition has a shelf life for a period in a range of 12 months to 24 months.
13. A process for preparing the composition as claimed in any one of the claims 1 to 3, said process comprising:
a. obtaining dodecanolide;
b. obtaining thymol;
c. obtaining at least one water phase component or at least one oil
phase component; and
d. contacting dodecanolide, thymol, the at least one water phase
component or the at least one oil phase component to obtain the
composition.
14. A process for preparing the composition as claimed in any one of the claims
1 and 4, said process comprising:
a. heating the at least one water phase component to a temperature in range of 65-75°C for a time period in a range of 25-35 minutes, to obtain a heated water phase component, and blending the heated

water phase component at a speed in a range of 200-400 rpm for a time period in a range of 10-20 minutes to obtain a first mixture;
b. heating the at least one oil phase component at a temperature in a
range of 60-80°C for a time period in a range of 25-35 minutes to
obtain, to obtain a heated oil phase component, and blending the
heated oil phase component at a speed of 300-500 rpm for a time
period in a range of 20-40 minutes to obtain a second mixture;
c. contacting the first mixture and the second mixture at a speed in a
range of 400-600 rpm for a time period in range of 2-40 minutes to
obtain a third mixture; and
d. contacting dodecanolide, thymol, the at least one solvent and the
third mixture at a speed in a range of 400-600 rpm for a time period
in a range of 40-50 minutes at a temperature in a range of 35-40°C,
to obtain the composition.
15. The process as claimed in any one of the claims 13 to 14, wherein the process further comprises at least one solvent having a weight percentage in a range of 10-98% with respect to the composition.
16. The process as claimed in claim 15, wherein the at least one solvent is selected from a group consisting of water, methanol, ethanol, dimethyl sulfoxide (DMSO), and combinations thereof.