FIELD OF INVENTION
[001] The present disclosure relates to the field of antifouling compounds in general, and in particular, relates to a novel composition comprising ascorbic acid and gallic acid for inhibiting biofilm formation.
BACKGROUND OF INVENTION
[002] Bacterial biofilms are aggregations of microbial cells which are embedded in a polysaccharide matrix. These biofilms allow bacteria to adhere to both biological and non-biological surfaces. Once within the biofilm matrix, the survival rates and ability to resist harsh environmental conditions of bacterial cells is considerably enhanced as the physiology and physiological responses of these cells is modified (Science against Microbial Pathogens: Communicating Current Research and Technological Advances; Esperanza Cortes et al., 2011; Vol 2; 693-1348). For instance, biofilms comprising sessile bacterial communities can often evolve to include planktonic forms, which are more resistant to antibiotics. [003] The polysaccharide matrix within which bacterial communities are embedded comprises extracellular polymeric substances (EPS) which are secreted by the cells themselves. The EPS is primarily comprised of a matrix of polysaccharides, lipids, lipopolysaccharides and other substances which act as a scaffold and also permit cell-cell communication.
[004] Bacterial biofilms are the causative agents of a number of diseases including dental caries. The primary etiological agent for causing human dental caries is the bacterium Streptococcus mutans. The bacterium can metabolize dietary carbohydrates to produce large amount of acid and can also survive in a low pH environment. Furthermore, S. mutans utilizes dietary sucrose to synthesize water soluble and insoluble glucans or polysaccharides which constitute dental plaque or biofilm.

[005] Biofilms retard the ability of antimicrobial substances, such as antibiotics, to diffuse within the matrix and reach bacterial cells, thereby imparting antimicrobial resistance to bacterial communities. Several different antimicrobials are used to control bacterial biofilms which include the use of bactericidal compounds, antibiotics, enzymes and bacteriophages, however, the effectiveness of these compounds is limited. Further, the excessive use of such bactericidal agents has led to the development of microbial communities that are resistant to these molecules. Thus, there is a need to find alternative compounds to inhibit the formation of bacterial biofilms.
SUMMARY OF INVENTION
[006] In an aspect of the present disclosure, there is provided a composition
comprising: (a) ascorbic acid; and (b) gallic acid, wherein gallic acid to ascorbic acid
w/w ratio in said composition is in the range of 1:1.5-1:4.
[007] In an aspect of the present disclosure, there is provided a method for preparing
a composition comprising a 1:1.5-1:4 w/w ratio of gallic acid: ascorbic acid, said
method comprising the steps of:
(a) obtaining ascorbic acid; (b) obtaining gallic acid; and (c) contacting ascorbic acid
and gallic acid to obtain said composition.
[008] In an aspect of the present disclosure, there is provided a method for
inhibiting bacterial biofilm formation, said method comprising:
(a) obtaining a composition comprising a 1:1.5-1:4 w/w ratio of gallic acid: ascorbic
acid; (b) contacting said composition with bacteria capable of biofilm formation,
wherein said composition inhibits bacterial biofilm formation.
[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 shows percentage control of biofilm formation by the individual
actives, ascorbic acid and gallic acid at different weight percentages, in accordance
with an embodiment of the present disclosure.
[0012] Figure 2 shows the percentage inhibition of biofilm formation by the
combination of ascorbic acid and gallic acid at different weight percentages.
[0013] Figure 3 shows the synergistic effect of a composition comprising a 1:4 w/w
ratio of gallic acid: ascorbic acid.
DETAILED DESCRIPTION OF THE INVENTION
[0014] The present disclosure relates to a composition comprising gallic acid and
ascorbic acid which at specific ratios and weight percentages demonstrates a
synergistic effect in the inhibition of biofilm formation.
[0015] 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
[0016] 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.
[0017] 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.
[0018] 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".
[0019] 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.
[0020] The term "including" is used to mean "including but not limited to".
"Including" and "including but not limited to" are used interchangeably.
[0021] Ascorbic acid, also known as Vitamin C, is an essential nutrient found in
many foods and is used as a dietary supplement.
[0022] Gallic acid, also known as trihydroxybenzoic acid, is found both free and as
part of hydrolyzable tannins.
[0023] 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.
[0024] The formation of bacterial biofilms is a major cause of disease. One of the
diseases caused by such biofilms is dental caries, and among the bacterial species
causing this disease is S. mutans. Most bacterial species have developed resistance

against most commonly used bactericidal compounds. This disclosure provides an
alternative solution to this problem in the form of a novel composition comprising
gallic acid and ascorbic acid which at specific w/w ratios, significantly inhibits
biofilm formation and can thus be used for the treatment of dental caries.
[0025] 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.
[0026] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) ascorbic acid; and (b) gallic acid, wherein
gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:1.5 - 1:4.
[0027] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) ascorbic acid; and (b) gallic acid, at a w/w ratio in the range of 1:1.5 -
1:4, wherein said composition is used to treat dental caries.
[0028] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) ascorbic acid; and (b) gallic acid, wherein
gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:2.5-l :4.
[0029] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) ascorbic acid; and (b) gallic acid, wherein
gallic acid to ascorbic acid w/w ratio in said composition is 1:4.
[0030] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) gallic acid; and (b) ascorbic acid, at a w/w ratio of 1:4, wherein said
composition is used to treat dental caries.
[0031] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) gallic acid having a weight percentage in the
range of 0.0064%-0.0512%; and (b) ascorbic acid, wherein gallic acid to ascorbic acid
w/w ratio in said composition is in the range of 1:1.5-1:4.

[0032] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) gallic acid having a weight percentage of
0.0128%; and (b) ascorbic acid, wherein gallic acid to ascorbic acid w/w ratio in said
composition is in the range of 1:1.5-1:4.
[0033] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) gallic acid having a weight percentage in the
range of 0.0064%-0.0512%; and (b) ascorbic acid, wherein gallic acid to ascorbic acid
w/w ratio in said composition is in the range of 1:2.5-l :4.
[0034] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) gallic acid having a weight percentage in the
range of 0.0064%-0.0512%; and (b) ascorbic acid, wherein gallic acid to ascorbic acid
w/w ratio in said composition is 1:4.
[0035] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) gallic acid having a weight percentage of
0.0128%; and (b) ascorbic acid, wherein gallic acid to ascorbic acid w/w ratio in said
composition is 1:4.
[0036] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) gallic acid; and (b) ascorbic acid having a
weight percentage in the range of 0.0064%-0.0512% wherein gallic acid to ascorbic
acid w/w ratio in said composition is in the range of 1:1.5-1:4.
[0037] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) gallic acid; and (b) ascorbic acid having a
weight percentage of 0.0128% wherein gallic acid to ascorbic acid w/w ratio in said
composition is in the range of 1:1.5-1:4.
[0038] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) gallic acid; and (b) ascorbic acid having a
weight percentage in the range of 0.0064%-0.0512%, wherein gallic acid to ascorbic
acid w/w ratio in said composition is in the range of 1:2.5-l :4.

[0039] In an embodiment of the present disclosure, there is provided a composition to inhibit biofilm formation comprising: (a) gallic acid; and (b) ascorbic acid having a weight percentage of 0.0128%, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:2.5-l :4.
[0040] In an embodiment of the present disclosure, there is provided a composition to inhibit biofilm formation comprising: (a) gallic acid; and (b) ascorbic acid having a weight percentage in the range of 0.0064% - 0.0512%>, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4.
[0041] In an embodiment of the present disclosure, there is provided a composition to inhibit biofilm formation comprising: (a) gallic acid; and (b) ascorbic acid having a weight percentage of 0.0128%), wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4.
[0042] In an embodiment of the present disclosure, there is provided a composition to inhibit biofilm formation comprising: (a) gallic acid having a weight percentage in the range of 0.0064%>-0.0512%>; and (b) ascorbic acid having a weight percentage in the range of 0.0064%>-0.0512%>, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:1.5-1:4.
[0043] In an embodiment of the present disclosure, there is provided a composition to treat dental caries comprising: (a) gallic acid having a weight percentage in the range of 0.0064%>-0.0512%>; and (b) ascorbic acid having a weight percentage in the range of 0.0064%>-0.0512%>, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:1.5-1:4
[0044] In an embodiment of the present disclosure, there is provided a composition to inhibit biofilm formation comprising: (a) gallic acid having a weight percentage of 0.0128%>; and (b) ascorbic acid having a weight percentage of 0.0128%>, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:1.5-1:4. [0045] In an embodiment of the present disclosure, there is provided a composition to treat dental caries comprising: (a) gallic acid having a weight percentage of 0.0128%>;

and (b) ascorbic acid having a weight percentage of 0.0128%, wherein gallic acid to
ascorbic acid w/w ratio in said composition is in the range of 1:1.5-1:4.
[0046] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) gallic acid having a weight percentage in the
range of 0.0064%-0.0512%; and (b) ascorbic acid having a weight percentage in the
range of 0.0064%-0.0512%, wherein gallic acid to ascorbic acid w/w ratio in said
composition is in the range of 1:2.5-1:4.
[0047] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) gallic acid having a weight percentage of
0.0128%; and (b) ascorbic acid having a weight percentage of 0.0128%, wherein
gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:2.5-l :4.
[0048] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) gallic acid having a weight percentage in the
range of 0.0064%-0.0512%; and (b) ascorbic acid having weight percentage in the
range of 0.0064%-0.0512%, wherein gallic acid to ascorbic acid w/w ratio in said
composition is 1:4.
[0049] In an embodiment of the present disclosure, there is provided a composition to
treat dental caries comprising: (a) gallic acid having a weight percentage in the range
of 0.0064%-0.0512%; and (b) ascorbic acid having weight percentage in the range of
0.0064%-0.0512%, wherein gallic acid to ascorbic acid w/w ratio in said composition
is 1:4.
[0050] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) gallic acid having a weight percentage of
0.0128%; and (b) ascorbic acid having weight percentage of 0.0128%), wherein gallic
acid to ascorbic acid w/w ratio in said composition is 1:4.
[0051] In an embodiment of the present disclosure, there is provided a composition to
treat dental caries comprising: (a) gallic acid having a weight percentage in the range

of 0.0128%; and (b) ascorbic acid having weight percentage of 0.0128%, wherein
gallic acid to ascorbic acid w/w ratio in said composition is 1:4.
[0052] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) gallic acid; (b) ascorbic acid; (c) at least one
carrier, (d) at least one diluent, and (e) at least one excipient, wherein gallic acid to
ascorbic acid w/w ratio in said composition is in the range of 1:1.5-1:4.
[0053] In an embodiment of the present disclosure, there is provided a composition to
treat dental caries comprising: (a) gallic acid; (b) ascorbic acid; (c) at least one carrier,
(d) at least one diluent, and (e) at least one excipient, wherein gallic acid to ascorbic
acid w/w ratio in said composition is in the range of 1:1.5-1:4.
[0054] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) gallic acid; (b) ascorbic acid; (c) at least one
carrier, (d) at least one diluent, and (e) at least one excipient, wherein gallic acid to
ascorbic acid w/w ratio in said composition is in the range of 1:2.5-l :4.
[0055] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) gallic acid; (b) ascorbic acid; and (c) at least
one carrier, (d) at least one diluent, and (e) at least one excipient, wherein gallic acid
to ascorbic acid w/w ratio in said composition is 1:4.
[0056] In an embodiment of the present disclosure, there is provided a composition to
treat dental caries comprising: (a) gallic acid; (b) ascorbic acid; and (c) at least one
carrier, (d) at least one diluent, and (e) at least one excipient, wherein gallic acid to
ascorbic acid w/w ratio in said composition is 1:4.
[0057] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) gallic acid having a weight percentage in the
range 0.0064%-0.0512%; (b) ascorbic acid; (c) at least one carrier, (d) at least one
diluent, and (e) at least one excipient, wherein gallic acid to ascorbic acid w/w ratio in
said composition is in the range of 1:1.5-1:4.

[0058] In an embodiment of the present disclosure, there is provided a composition to inhibit biofilm formation comprising: (a) gallic acid having a weight percentage in the range of 0.0064%-0.0512%; (b) ascorbic acid; (c) at least one carrier, (d) at least one diluent, and (e) at least one excipient, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:2.5-1:4.
[0059] In an embodiment of the present disclosure, there is provided a composition to inhibit biofilm formation comprising: (a) gallic acid having a weight percentage in the range of 0.0064%-0.0512%; (b) ascorbic acid; (c) at least one carrier, (d) at least one diluent, and (e) at least one excipient, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4.
[0060] In an embodiment of the present disclosure, there is provided a composition to inhibit biofilm formation comprising: (a) gallic acid having a weight percentage of 0.0128%; (b) ascorbic acid; (c) at least one carrier, (d) at least one diluent, and (e) at least one excipient, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:1.5-1:4.
[0061] In an embodiment of the present disclosure, there is provided a composition to inhibit biofilm formation comprising: (a) gallic acid having a weight percentage of 0.0128%; (b) ascorbic acid; (c) at least one carrier, (d) at least one diluent, and (e) at least one excipient, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:2.5-1:4.
[0062] In an embodiment of the present disclosure, there is provided a composition to inhibit biofilm formation comprising: (a) gallic acid having a weight percentage of 0.0128%; (b) ascorbic acid; (c) at least one carrier, (d) at least one diluent, and (e) at least one excipient wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4.
[0063] In an embodiment of the present disclosure, there is provided a composition to inhibit biofilm formation comprising: (a) gallic acid; (b) ascorbic acid having a weight percentage in the range of 0.0064%-0.0512%; (c) at least one carrier, (d) at least one

diluent, and (e) at least one excipient, wherein gallic acid to ascorbic acid w/w ratio in
said composition is in the range of 1:1.5-1:4.
[0064] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) gallic acid; (b) ascorbic acid having a weight
percentage in the range of 0.0064%-0.0512%; (c) at least one carrier, (d) at least one
diluent, and (e) at least one excipient, wherein gallic acid to ascorbic acid w/w ratio in
said composition is in the range of 1:2.5-l :4.
[0065] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) gallic acid; (b) ascorbic acid having a weight
percentage in the range of 0.0064%-0.0512%; (c) at least one carrier, (d) at least one
diluent, and (e) at least one excipient, wherein gallic acid to ascorbic acid w/w ratio in
said composition is in the range of 1:4.
[0066] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) gallic acid; (b) ascorbic acid having a weight
percentage of 0.0128%; (c) at least one carrier, (d) at least one diluent, and (e) at least
one excipient, wherein gallic acid to ascorbic acid w/w ratio in said composition is in
the range of 1:1.5-1:4.
[0067] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) gallic acid; (b) ascorbic acid having a weight
percentage of 0.0128%; (c) at least one carrier, (d) at least one diluent, and (e) at least
one excipient, wherein gallic acid to ascorbic acid w/w ratio in said composition is in
the range of 1:2.5-1:4.
[0068] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) gallic acid; (b) ascorbic acid having a weight
percentage of 0.0128%; (c) at least one carrier, (d) at least one diluent, and (e) at least
one excipient, wherein gallic acid to ascorbic acid w/w ratio in said composition is
1:4.

[0069] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) gallic acid having a weight percentage in the
range of 0.0064%-0.0512%; (b) ascorbic acid having a weight percentage in the range
of 0.0064%-0.0512%; (c) at least one carrier, (d) at least one diluent, and (e) at least
one excipient, wherein gallic acid to ascorbic acid w/w ratio in said composition is in
the range of 1:1.5-1:4.
[0070] In an embodiment of the present disclosure, there is provided a composition to
treat dental caries comprising: (a) gallic acid having a weight percentage in the range
of 0.0064%-0.0512%; (b) ascorbic acid having a weight percentage in the range of
0.0064%-0.0512%; (c) at least one carrier, (d) at least one diluent, and (e) at least one
excipient, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the
range of 1:1.5-1:4.
[0071] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) gallic acid having a weight percentage in the
range of 0.0064%-0.0512%; (b) ascorbic acid having weight percentage in the range
of 0.0064%-0.0512%; (c) at least one carrier, (d) at least one diluent, and (e) at least
one excipient, wherein gallic acid to ascorbic acid w/w ratio in said composition is
1:2.5:1:4.
[0072] In an embodiment of the present disclosure, there is provided a composition to
inhibit biofilm formation comprising: (a) gallic acid having a weight percentage in the
range of 0.0064%-0.0512%; (b) ascorbic acid having weight percentage in the range
of 0.0064%-0.0512%; (c) at least one carrier, (d) at least one diluent, and (e) at least
one excipient, wherein gallic acid to ascorbic acid w/w ratio in said composition is
1:4.
[0073] In an embodiment of the present disclosure, there is provided a composition to
treat dental caries comprising: (a) gallic acid having a weight percentage in the range
of 0.0064%-0.0512%; (b) ascorbic acid having weight percentage in the range of

0.0064%-0.0512%; (c) at least one carrier, (d) at least one diluent, and (e) at least one excipient, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4. [0074] In an embodiment of the present disclosure, there is provided a composition to inhibit biofilm formation comprising: (a) gallic acid having a weight percentage of 0.0128%; (b) ascorbic acid having weight percentage of 0.0128%; (c) at least one carrier, (d) at least one diluent, and (e) at least one excipient, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:1.5-1:4. [0075] In an embodiment of the present disclosure, there is provided a composition to treat dental caries comprising: (a) gallic acid having a weight percentage of 0.0128%; (b) ascorbic acid having weight percentage of 0.0128%; (c) at least one carrier, (d) at least one diluent, and (e) at least one excipient, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:1.5-1:4.
[0076] In an embodiment of the present disclosure, there is provided a composition to inhibit biofilm formation comprising: (a) gallic acid having a weight percentage of 0.0128%; (b) ascorbic acid having weight percentage of 0.0128%; (c) at least one carrier, (d) at least one diluent, and (e) at least one excipient, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:2.5-1:4.
[0077] In an embodiment of the present disclosure, there is provided a composition to inhibit biofilm formation comprising: (a) gallic acid having a weight percentage of 0.0128%; (b) ascorbic acid having weight percentage of 0.0128%; (c) at least one carrier, (d) at least one diluent, and (e) at least one excipient, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4.
[0078] In an embodiment of the present disclosure, there is provided a composition to treat dental caries comprising: (a) gallic acid having a weight percentage of 0.0128%; (b) ascorbic acid having weight percentage of 0.0128%; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4.

[0079] In an embodiment of the present disclosure, there is provided a method of
preparing a composition to inhibit biofilm formation comprising: (a) obtaining
ascorbic acid; (b) obtaining gallic acid; and (c) contacting ascorbic acid and gallic
acid to obtain said composition.
[0080] In an embodiment of the present disclosure, there is provided a method of
preparing a composition to inhibit biofilm formation comprising: (a) obtaining
ascorbic acid; (b) obtaining gallic acid; and (c) contacting ascorbic acid and gallic
acid to obtain said composition, wherein gallic acid to ascorbic acid w/w ratio in said
composition is in the range of 1:1.5-1:4.
[0081] In an embodiment of the present disclosure, there is provided a method of
preparing a composition to inhibit biofilm formation comprising: (a) obtaining
ascorbic acid; (b) obtaining gallic acid; and (c) contacting ascorbic acid and gallic
acid to obtain said composition, wherein gallic acid to ascorbic acid w/w ratio in said
composition is in the range of 1:2.5-l :4.
[0082] In an embodiment of the present disclosure, there is provided a method of
preparing a composition to inhibit biofilm formation comprising: (a) obtaining
ascorbic acid; (b) obtaining gallic acid; and (c) contacting ascorbic acid and gallic
acid to obtain said composition, wherein gallic acid to ascorbic acid w/w ratio in said
composition is in the range of 1:4.
[0083] In an embodiment of the present disclosure, there is provided a method of
preparing a composition as described herein, wherein said composition comprises: (a)
gallic acid having a weight percentage in the range of 0.0064%-0.0512%; and (b)
ascorbic acid, wherein gallic acid to ascorbic acid w/w ratio in said composition is in
the range of 1:1.5-1:4.
[0084] In an embodiment of the present disclosure, there is provided a method of
preparing a composition as described herein, wherein said composition comprises: (a)
gallic acid having a weight percentage in the range of 0.0064%-0.0512%; and (b)

ascorbic acid, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:2.5-1:4.
[0085] In an embodiment of the present disclosure, there is provided a method of preparing a composition as described herein, wherein said composition comprises: (a) gallic acid having a weight percentage in the range of 0.0064%-0.0512%; and (b) ascorbic acid, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:4.
[0086] In an embodiment of the present disclosure, there is provided a method of preparing a composition as described herein, wherein said composition comprises: (a) gallic acid having a weight percentage of 0.0128%; and (b) ascorbic acid, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:1.5-1:4. [0087] In an embodiment of the present disclosure, there is provided a method of preparing a composition as described herein, wherein said composition comprises: (a) gallic acid having a weight percentage of 0.0128%; and (b) ascorbic acid, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:2.5-l :4. [0088] In an embodiment of the present disclosure, there is provided a method of preparing a composition as described herein, wherein said composition comprises: (a) gallic acid having a weight percentage of 0.0128%; and (b) ascorbic acid, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4. [0089] In an embodiment of the present disclosure, there is provided a method of preparing a composition as described herein, wherein said composition comprises: (a) gallic acid; and (b) ascorbic acid having a weight percentage in the range of 0.0064%-0.0512%), wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:1.5-1:4.
[0090] In an embodiment of the present disclosure, there is provided a method of preparing a composition described herein, wherein said composition comprises: (a) gallic acid; and (b) ascorbic acid having a weight percentage in the range of 0.0064%>-

0.0512%, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:2.5-1:4.
[0091] In an embodiment of the present disclosure, there is provided a method of preparing a composition described herein, wherein said composition comprises: (a) gallic acid; and (b) ascorbic acid having a weight percentage in the range of 0.0064%-0.0512%), wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:4.
[0092] In an embodiment of the present disclosure, there is provided a method of preparing a composition as described herein, wherein said composition comprises: (a) gallic acid; and (b) ascorbic acid having a weight percentage of 0.0128%>, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:1.5-1:4. [0093] In an embodiment of the present disclosure, there is provided a method of preparing a composition as described herein, wherein said composition comprises: (a) gallic acid; and (b) ascorbic acid having a weight percentage of 0.0128%>, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:2.5-l :4. [0094] In an embodiment of the present disclosure, there is provided a method of preparing a composition as described herein, wherein said composition comprises: (a) gallic acid; and (b) ascorbic acid having a weight percentage of 0.0128%>, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4 [0095] In an embodiment of the present disclosure, there is provided a method of preparing a composition described herein, wherein said composition comprises: (a) gallic acid having a weight percentage in the range of 0.0064%>-0.0512%>; and (b) ascorbic acid having weight percentage in the range of 0.0064%>-0.0512%>, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:1.5-1:4. [0096] In an embodiment of the present disclosure, there is provided a method of preparing a composition as described herein, wherein said composition comprises: (a) gallic acid having a weight percentage in the range of 0.0064%>-0.0512%>; and (b)

ascorbic acid having weight percentage in the range of 0.0064%-0.0512%, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:2.5-l :4. [0097] In an embodiment of the present disclosure, there is provided a method of preparing a composition as described herein, wherein said composition comprises: (a) gallic acid having a weight percentage in the range of 0.0064%-0.0512%; and (b) ascorbic acid having a weight percentage in the range of 0.0064%-0.0512%, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4 [0098] In an embodiment of the present disclosure, there is provided a method of preparing a composition as described herein, wherein said composition comprises: (a) gallic acid having a weight percentage of 0.0128%; and (b) ascorbic acid having weight percentage of 0.0128%, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:1.5-1:4.
[0099] In an embodiment of the present disclosure, there is provided a method of preparing a composition as described herein, wherein said composition comprises: (a) gallic acid having a weight percentage of 0.0128%; and (b) ascorbic acid having weight percentage of 0.0128%), wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:2.5-1:4.
[00100] In an embodiment of the present disclosure, there is provided a method of preparing a composition described herein, wherein said composition comprises: (a) gallic acid having a weight percentage of 0.0128%>; and (b) ascorbic acid having weight percentage of 0.0128%), wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4.
[00101] In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit biofilm formation comprising: (a) obtaining ascorbic acid; (b) obtaining gallic acid; (c) obtaining at least one carrier, at least one diluent, at least one excipient, and combinations thereof; and (d) contacting ascorbic acid, gallic acid and carrier, diluent, excipient and combinations thereof, to obtain said composition.

[00102] In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit biofilm formation comprising: (a) obtaining ascorbic acid; (b) obtaining gallic acid; (c) obtaining at least one carrier, at least one diluent, at least one excipient, and combinations thereof; and (d) contacting ascorbic acid, gallic acid and carrier, diluent, excipient and combinations thereof, to obtain said composition, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:1.5-1:4.
[00103] In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit biofilm formation comprising: (a) obtaining ascorbic acid; (b) obtaining gallic acid; (c) obtaining at least one carrier, at least one diluent, at least one excipient, and combinations thereof; and (d) contacting ascorbic acid, gallic acid and carrier, diluent, excipient and combinations thereof, to obtain said composition, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:2.5-1:4.
[00104] In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit biofilm formation comprising: (a) obtaining ascorbic acid; (b) obtaining gallic acid; (c) obtaining at least one carrier, at least one diluent, at least one excipient, and combinations thereof; and (d) contacting ascorbic acid, gallic acid and carrier, diluent, excipient and combinations thereof, to obtain said composition, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:4.
[00105] In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit biofilm formation comprising: (a) obtaining gallic acid having a weight percentage in the range of 0.0064%-0.0512%; (b) obtaining ascorbic acid; (c) obtaining at least one carrier, at least one diluent, at least one excipient, and combinations thereof; and (d) contacting ascorbic acid, gallic acid and carrier, diluent, excipient and combinations thereof, to obtain said composition,

wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:1.5-1:4.
[00106] In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit biofilm formation comprising: (a) gallic acid having a weight percentage in the range of 0.0064%-0.0512%; (b) ascorbic acid; (c) obtaining at least one carrier, at least one diluent, at least one excipient, and combinations thereof; and (d) contacting ascorbic acid, gallic acid and carrier, diluent, excipient and combinations thereof, to obtain said composition, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:2.5-l :4. [00107] In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit biofilm formation comprising: (a) gallic acid having a weight percentage in the range of 0.0064%-0.0512%; (b) ascorbic acid; (c) obtaining at least one carrier, at least one diluent, at least one excipient, and combinations thereof; and (d) contacting ascorbic acid, gallic acid and carrier, diluent, excipient and combinations thereof, to obtain said composition, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4.
[00108] In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit biofilm formation comprising: (a) gallic acid having a weight percentage of 0.0128%; (b) ascorbic acid; (c) obtaining at least one carrier, at least one diluent, at least one excipient, and combinations thereof; and (d) contacting ascorbic acid, gallic acid and carrier, diluent, excipient and combinations thereof, to obtain said composition, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:1.5-1:4.
[00109] In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit biofilm formation comprising: (a) gallic acid having a weight percentage of 0.0128%; (b) ascorbic acid; (c) obtaining at least one carrier, at least one diluent, at least one excipient, and combinations thereof; and (d) contacting ascorbic acid, gallic acid and carrier, diluent, excipient and combinations

thereof, to obtain said composition, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:2.5-1:4.
[OOllOJIn an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit biofilm formation comprising: (a) gallic acid having a weight percentage of 0.0128%; (b) ascorbic acid; (c) obtaining at least one carrier, at least one diluent, at least one excipient, and combinations thereof; and (d) contacting ascorbic acid, gallic acid and carrier, diluent, excipient and combinations thereof, to obtain said composition, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:4.
[00111]In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit biofilm formation comprising: (a) gallic acid; (b) ascorbic acid having a weight percentage in the range of 0.0064%-0.0512%; (c) obtaining at least one carrier, at least one diluent, at least one excipient, and combinations thereof; and (d) contacting ascorbic acid, gallic acid and carrier, diluent, excipient and combinations thereof, to obtain said composition, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:1.5-1:4. [00112]In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit biofilm formation comprising: (a) gallic acid; and (b) ascorbic acid having a weight percentage in the range of 0.0064%-0.0512%, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:2.5-1:4.
[00113]In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit biofilm formation comprising: (a) gallic acid; (b) ascorbic acid having a weight percentage in the range of 0.0064%-0.0512%; (c) obtaining at least one carrier, at least one diluent, at least one excipient, and combinations thereof; and (d) contacting ascorbic acid, gallic acid and carrier, diluent, excipient and combinations thereof, to obtain said composition, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4.

[00114]In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit biofilm formation comprising: (a) gallic acid; (b) ascorbic acid having a weight percentage of 0.0128%; (c) obtaining at least one carrier, at least one diluent, at least one excipient, and combinations thereof; and (d) contacting ascorbic acid, gallic acid and carrier, diluent, excipient and combinations thereof, to obtain said composition, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:1.5-1:4.
[00115]In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit biofilm formation comprising: (a) gallic acid; (b) ascorbic acid having a weight percentage of 0.0128%; (c) obtaining at least one carrier, at least one diluent, at least one excipient, and combinations thereof; and (d) contacting ascorbic acid, gallic acid and carrier, diluent, excipient and combinations thereof, to obtain said composition, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:2.5-1:4.
[00116]In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit biofilm formation comprising: (a) gallic acid; (b) ascorbic acid having a weight percentage of 0.0128%; (c) obtaining at least one carrier, at least one diluent, at least one excipient, and combinations thereof; and (d) contacting ascorbic acid, gallic acid and carrier, diluent, excipient and combinations thereof, to obtain said composition, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:4
[00117]In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit biofilm formation comprising: (a) gallic acid having a weight percentage in the range of 0.0064%-0.0512%; (b) ascorbic acid having weight percentage in the range of 0.0064%-0.0512%; (c) obtaining at least one carrier, at least one diluent, at least one excipient, and combinations thereof; and (d) contacting ascorbic acid, gallic acid and carrier, diluent, excipient and combinations

thereof, to obtain said composition, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:1.5-1:4.
[00118]In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit biofilm formation comprising: (a) gallic acid having a weight percentage of 0.0064%-0.0512%; (b) ascorbic acid having weight percentage of 0.0064%-0.0512%; (c) obtaining at least one carrier, at least one diluent, at least one excipient, and combinations thereof; and (d) contacting ascorbic acid, gallic acid and carrier, diluent, excipient and combinations thereof, to obtain said composition, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:2.5-1:4.
[00119]In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit biofilm formation comprising: (a) gallic acid having a weight percentage in the range of 0.0064%-0.0512%; (b) ascorbic acid having a weight percentage in the range of 0.0064%-0.0512%; (c) obtaining at least one carrier, at least one diluent, at least one excipient, and combinations thereof; and (d) contacting ascorbic acid, gallic acid and carrier, diluent, excipient and combinations thereof, to obtain said composition, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4
[00120] In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit biofilm formation comprising: (a) gallic acid having a weight percentage of 0.0128%; (b) ascorbic acid having weight percentage of 0.0128%; (c) obtaining at least one carrier, at least one diluent, at least one excipient, and combinations thereof; and (d) contacting ascorbic acid, gallic acid and carrier, diluent, excipient and combinations thereof, to obtain said composition, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:1.5-1:4.
[00121] In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit biofilm formation comprising: (a) gallic acid

having a weight percentage of 0.0128%; (b) ascorbic acid having weight percentage of 0.0128%; (c) obtaining at least one carrier, at least one diluent, at least one excipient, and combinations thereof; and (d) contacting ascorbic acid, gallic acid and carrier, diluent, excipient and combinations thereof, to obtain said composition, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:2.5-1:4. [00122] In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit biofilm formation comprising: (a) gallic acid having a weight percentage of 0.0128%; (b) ascorbic acid having weight percentage of 0.0128%; (c) obtaining at least one carrier, at least one diluent, at least one excipient, and combinations thereof; and (d) contacting ascorbic acid, gallic acid and carrier, diluent, excipient and combinations thereof, to obtain said composition, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4. [00123] In an embodiment of the present disclosure, there is provided a method for inhibiting bacterial biofilm formation comprising:
(a) obtaining a composition comprising; (i) gallic acid, and (ii) ascorbic acid, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:1.5-1:4; and
(b) contacting said composition with bacteria capable of biofilm formation, including but not limited to Streptococcus mutans; wherein said composition inhibits bacterial biofilm formation.
[00124] In an embodiment of the present disclosure, there is provided a method for inhibiting bacterial biofilm formation comprising:
(a) obtaining a composition comprising; (i) gallic acid, and (ii) ascorbic acid, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4; and
(b) contacting said composition with bacteria capable of biofilm formation, including but not limited to Streptococcus mutans; wherein said composition inhibits bacterial biofilm formation.
[00125] In an embodiment of the present disclosure, there is provided a method for inhibiting bacterial biofilm formation comprising:

(a) obtaining a composition comprising; (i) gallic acid having a weight percentage of 0.0064%-0.0512%, and (ii) ascorbic acid, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:1.5-1:4; and
(b) contacting said composition with bacteria capable of biofilm formation, including but not limited to Streptococcus mutans; wherein said composition inhibits bacterial biofilm formation.
[00126] In an embodiment of the present disclosure , there is provided a method for inhibiting bacterial biofilm formation comprising:
(a) obtaining a composition comprising; (i) gallic acid, and (ii) ascorbic acid having a weight percentage of 0.0064%-0.0512%, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:1.5-1:4; and (b) contacting said composition with bacteria capable of biofilm formation, including but not limited to Streptococcus mutans; wherein said composition inhibits bacterial biofilm formation. [00127] In an embodiment of the present disclosure, there is provided a method for inhibiting bacterial biofilm formation comprising:
(a) obtaining a composition comprising; (i) gallic acid having a weight percentage of 0.0064%-0.0512%, and (ii) ascorbic acid, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4; and (b) contacting said composition with bacteria capable of biofilm formation, including but not limited to Streptococcus mutans; wherein said composition inhibits bacterial biofilm formation.
[00128] In an embodiment of the present disclosure, there is provided a method for inhibiting bacterial biofilm formation comprising:
(a) obtaining a composition comprising; (i) gallic acid, and (ii) ascorbic acid having a weight percentage of 0.0064%-0.0512%, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4; and (b) contacting said composition with bacteria capable of biofilm formation, including but not limited to Streptococcus mutans; wherein said composition inhibits bacterial biofilm formation.

[00129] In an embodiment of the present disclosure, there is provided a method for inhibiting bacterial biofilm formation comprising:
(a) obtaining a composition comprising; (i) gallic acid having a weight percentage of 0.0064%-0.0512%, and (ii) ascorbic acid having a weight percentage of 0.0064%-0.0512%, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:1.5-1:4; and (b) contacting said composition with bacteria capable of biofilm formation, including but not limited to Streptococcus mutans; wherein said composition inhibits bacterial biofilm formation.
[00130] In an embodiment of the present disclosure, there is provided a method for inhibiting bacterial biofilm formation comprising:
(a) obtaining a composition comprising; (i) gallic acid having a weight percentage of 0.0064%-0.0512%, and (ii) ascorbic acid having a weight percentage of 0.0064%-0.0512%), wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4; and (b) contacting said composition with bacteria capable of biofilm formation, including but not limited to Streptococcus mutans; wherein said composition inhibits bacterial biofilm formation.
[00131] In an embodiment of the present disclosure, there is provided a method for inhibiting bacterial biofilm formation comprising:
(a) obtaining a composition comprising; (i) gallic acid having a weight percentage of 0.0128%), and (ii) ascorbic acid having a weight percentage of 0.0128%>, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:1.5-1:4; and
(b) contacting said composition with bacteria capable of biofilm formation, including but not limited to Streptococcus mutans; wherein said composition inhibits bacterial biofilm formation.
[00132] In an embodiment of the present disclosure, there is provided a method for inhibiting bacterial biofilm formation comprising:
(a) obtaining a composition comprising; (i) gallic acid having a weight percentage of 0.0128%o, and (ii) ascorbic acid, wherein gallic acid to ascorbic acid w/w ratio in said

composition is 1:1.5-1:4; and (b) contacting said composition with bacteria capable of biofilm formation, including but not limited to Streptococcus mutans; wherein said composition inhibits bacterial biofilm formation.
[00133] In an embodiment of the present disclosure, there is provided a method for inhibiting bacterial biofilm formation comprising:
(a) obtaining a composition comprising; (i) gallic acid; and (ii) ascorbic acid having a weight percentage of 0.0128%, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:1.5-1:4; and (b) contacting said composition with bacteria capable of biofilm formation, including but not limited to Streptococcus mutans; wherein said composition inhibits bacterial biofilm formation.
[00134] In an embodiment of the present disclosure, there is provided a method for inhibiting bacterial biofilm formation comprising:
(a) obtaining a composition comprising; (i) gallic acid having a weight percentage of 0.0128%, and (ii) ascorbic acid having a weight percentage of 0.0128%, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4; and (b) contacting said composition with bacteria capable of biofilm formation, including but not limited to Streptococcus mutans; wherein said composition inhibits bacterial film formation. [00135] In an embodiment of the present disclosure , there is provided a method for inhibiting bacterial biofilm formation comprising:
(a) obtaining a composition comprising; (i) gallic acid, (ii) ascorbic acid, and (iii) at least one carrier, at least one, at least one diluent, at least one excipient, and combinations thereof, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:1.5-1:4; and (b) contacting said composition with bacteria capable of biofilm formation, including but not limited to Streptococcus mutans; wherein said composition inhibits bacterial biofilm formation.
[00136] In an embodiment of the present disclosure, there is provided a method for inhibiting bacterial biofilm formation comprising:

(a) obtaining a composition comprising; (i) gallic acid, (ii) ascorbic acid, and (iii) at least one carrier, at least one, at least one diluent, at least one excipient, and combinations thereof, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4; and (b) contacting said composition with bacteria capable of biofilm formation, including but not limited to Streptococcus mutans; wherein said composition inhibits bacterial biofilm formation.
[00137] In an embodiment of the present disclosure, there is provided a method for inhibiting bacterial biofilm formation comprising:
(a) obtaining a composition comprising; (i) gallic acid having a weight percentage of 0.0064%-0.0512%, (ii) ascorbic acid having a weight percentage of 0.0064%-0.0512%, and (iii) at least one carrier, at least one, at least one diluent, at least one excipient, and combinations thereof, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:1.5-1:4; and (b) contacting said composition with bacteria capable of biofilm formation, including but not limited to Streptococcus mutans; wherein said composition inhibits bacterial biofilm formation.
[00138] In an embodiment of the present disclosure, there is provided a method for inhibiting bacterial biofilm formation comprising:
(a) obtaining a composition comprising; (i) gallic acid having a weight percentage of 0.0064%-0.0512%, (ii) ascorbic acid, and (iii) at least one carrier, at least one, at least one diluent, at least one excipient, and combinations thereof, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:1.5-1:4; and (b) contacting said composition with bacteria capable of biofilm formation, including but not limited to Streptococcus mutans; wherein said composition inhibits bacterial biofilm formation. [00139] In an embodiment of the present disclosure, there is provided a method for inhibiting bacterial biofilm formation comprising:
(a) obtaining a composition comprising; (i) gallic acid having a weight percentage of 0.0064%-0.0512%, (ii) ascorbic acid having a weight percentage of 0.0064%-0.0512%), and (iii) at least one carrier, at least one, at least one diluent, at least one

excipient, and combinations thereof, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:1.5-1:4; and (b) contacting said composition with bacteria capable of biofilm formation, including but not limited to Streptococcus mutans; wherein said composition inhibits bacterial biofilm formation.
[00140] In an embodiment of the present disclosure , there is provided a method for inhibiting bacterial biofilm formation comprising:
(a) obtaining a composition comprising; (i) gallic acid having a weight percentage of 0.0064%-0.0512%, (ii) ascorbic acid having a weight percentage of 0.0064%-0.0512%, and (iii) at least one carrier, at least one, at least one diluent, at least one excipient, and combinations thereof, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4; and (b) contacting said composition with bacteria capable of biofilm formation, including but not limited to Streptococcus mutans; wherein said composition inhibits bacterial biofilm formation.
[00141] In an embodiment of the present disclosure, there is provided a method for inhibiting bacterial biofilm formation comprising:
(a) obtaining a composition comprising; (i) gallic acid having a weight percentage of 0.0064%-0.0512%, (ii) ascorbic acid, and (iii) at least one carrier, at least one, at least one diluent, at least one excipient, and combinations thereof, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4; and (b) contacting said composition with bacteria capable of biofilm formation, including but not limited to Streptococcus mutans; wherein said composition inhibits bacterial biofilm formation. [00142] In an embodiment of the present disclosure, there is provided a method for inhibiting bacterial biofilm formation comprising:
(a) obtaining a composition comprising; (i) gallic acid, (ii) ascorbic acid having a weight percentage of 0.0064%-0.0512%, and (iii) at least one carrier, at least one, at least one diluent, at least one excipient, and combinations thereof, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4; and (b) contacting said

composition with bacteria capable of biofilm formation, including but not limited to Streptococcus mutans; wherein said composition inhibits bacterial biofilm formation. [00143] In an embodiment of the present disclosure, there is provided a method for inhibiting bacterial biofilm formation comprising:
(a) obtaining a composition comprising; (i) gallic acid having a weight percentage of 0.0128%, (ii) ascorbic acid having a weight percentage of 0.0128%, and (iii) at least one carrier, at least one, at least one diluent, at least one excipient, and combinations thereof, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:1.5-1:4; and (b) contacting said composition with bacteria capable of biofilm formation, including but not limited to Streptococcus mutans; wherein said composition inhibits bacterial biofilm formation.
[00144] In an embodiment of the present disclosure , there is provided a method for inhibiting bacterial biofilm formation comprising:
(a) obtaining a composition comprising; (i) gallic acid having a weight percentage of 0.0128%), (ii) ascorbic acid, and (iii) at least one carrier, at least one, at least one diluent, at least one excipient, and combinations thereof, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:1.5-1:4; and (b) contacting said composition with bacteria capable of biofilm formation, including but not limited to Streptococcus mutans; wherein said composition inhibits bacterial biofilm formation. [00145] In an embodiment of the present disclosure , there is provided a method for inhibiting bacterial biofilm formation comprising:
(a) obtaining a composition comprising; (i) gallic acid having a weight percentage of 0.0128%), (ii) ascorbic acid having a weight percentage of 0.0128%>, and (iii) at least one carrier, at least one, at least one diluent, at least one excipient, and combinations thereof, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:1.5-1:4; and (b) contacting said composition with bacteria capable of biofilm formation, including but not limited to Streptococcus mutans; wherein said composition inhibits bacterial biofilm formation.

[00146] In an embodiment of the present disclosure , there is provided a method for inhibiting bacterial biofilm formation comprising:
(a) obtaining a composition comprising; (i) gallic acid having a weight percentage of 0.0128%, (ii) ascorbic acid having a weight percentage of 0.0128%, and (iii) at least one carrier, at least one, at least one diluent, at least one excipient, and combinations thereof, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4; and
(b) contacting said composition with bacteria capable of biofilm formation, including but not limited to Streptococcus mutans; wherein said composition inhibits bacterial biofilm formation.
[00147] In an embodiment of the present disclosure , there is provided a method for inhibiting bacterial biofilm formation comprising:
(a) obtaining a composition comprising; (i) gallic acid having a weight percentage of 0.0128%), (ii) ascorbic acid, and (iii) at least one carrier, at least one, at least one diluent, at least one excipient, and combinations thereof, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4; and (b) contacting said composition with bacteria capable of biofilm formation, including but not limited to Streptococcus mutans; wherein said composition inhibits bacterial biofilm formation. [00148] In an embodiment of the present disclosure , there is provided a method for inhibiting bacterial biofilm formation comprising:
(a) obtaining a composition comprising; (i) gallic acid, (ii) ascorbic acid having a weight percentage of 0.0128%>, and (iii) at least one carrier, at least one, at least one diluent, at least one excipient, and combinations thereof, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4; and (b) contacting said composition with bacteria capable of biofilm formation, including but not limited to Streptococcus mutans; wherein said composition inhibits bacterial biofilm formation. [00149]Although the subject matter has been described with reference to specific embodiments, this description is not meant to be construed in a limiting sense. Various modifications of the disclosed embodiments, as well as alternate

embodiments of the subject matter, will become apparent to persons skilled in the art upon reference to the description of the subject matter. It is therefore contemplated that such modifications can be made without departing from the spirit or scope of the present subject matter as defined.
EXAMPLES
[00150] 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 vary. Example 1
Material and Methods
Chemicals and materials: Ascorbic acid (Catalogue No. A4544-25G, Sigma Aldrich, MO, USA), Gallic acid (14590, Molychem, Mumbai, India), Sucrose (S0389-1KG, Sigma Aldrich, MO, USA), (Crystal Violet (Catalogue No. C0775-25G, Sigma Aldrich, MO, USA), glacial acetic acid (Catalogue No. 01074101, BIOSOLVE, Netherlands), Supplemented Tryptic Soy Agar (Catalogue No. 2722, ATCC, VA, USA), Brain Heart Infusion (BHI) Medium (Catalogue No. M210, HiMedia Laboratories, Mumbai, India), Costar 96-Well flat clear bottom tissue culture treated plates (Catalogue No. 3596, Corning, NY, USA), Oxoid 2.5L jar (Catalogue No. AG0025A, Thermo Scientific), Oxoid AnaeroGen 2.5L sachets (Catalogue No. AN0025A, Thermo Scientific). Stock solutions (100 mg/mL) of all actives was

prepared in cell culture grade DMSO. DMSO was used as a vehicle control in all experiments.
Bacterial strains and culture conditions: Streptococcus mutans (ATCC 25175) was used for the experiments disclosed herein. Overnight cultures of the bacterium were grown in BHI (Brain Heart Infusion) broth at 37 °C. Briefly, the bacterium was inoculated in 5.0 mL of BHI broth in a tightly capped sterile 15.0 mL polypropylene tube and placed under static condition for 18 h in an incubator set at 37 °C. Bacterial cultures were streaked on supplemented tryptic soy agar plates and the same were placed in an Oxoid 2.5L jar with Oxoid AnaeroGen 2.5L sachets in an incubator set at 37 °C. Colonies were isolated after 24 h and used for inoculating the BHI broth. Biofilm formation assay: To evaluate biofilm formation in the presence and absence of actives, S. mutans (106 CFU/mL) was grown in BHI broth supplemented with 0.05% sucrose in flat bottomed tissue culture treated 96-well plates. The plates were wrapped with plastic cling wrap and placed under static condition in an incubator set at 37 °C for 24 h. After 24 h, the medium with planktonic cells was removed by gently inverting the plate. The plate was completely immersed in a bath of tap water (2X) to wash off loosely bound cells and biofilm. The plate was inverted and tapped on folds of absorbent paper to get rid of excess liquids and the placed in an incubator set at 37 °C for 90 min. 100 uL of 0.01% crystal violet solution in water was added to the wells and the plate was incubated at room temperature for 60 min. The plate was inverted to discard the crystal violet solution and then completely immersed in a bath of tap water (3X) to wash off excess crystal violet and loosely attached bacterial cells and biofilm. The plate was inverted and tapped on folds of absorbent paper to get rid of excess liquids and placed in an incubator set at 37 °C for 60 min. 200 uL of 30% acetic acid in water was added to the wells and the plate was incubated at room temperature for 30 min. The contents of the wells were mixed well and 100 uL was transferred to a fresh flat clear bottom 96-well plate and absorbance measured at 590 nm on a spectrophotometer (MULTISKAN GO, Thermo Scientific).

Example 2
Inhibition of biofilm formation by gallic acid and ascorbic acid
[00151] In order to determine the concentrations at which gallic acid and ascorbic acid
were effective in isolation in inhibiting biofilm formation, the individual constituents
were tested at weight/weight percentages ranging from 0.0001% to 0.0512%. It was
found that the individual actives had the highest activity when each was present at a
concentration of 0.0128%) (Figure 1). At this concentration, ascorbic acid showed a
percent control of 86.8%>, while gallic acid showed a percent control of 93.7%. Below
the concentration of 0.0128%), the individual actives were not effective in inhibiting
biofilm formation, while concentrations above 0.0128%) did not show any marked
improvement in the inhibition of biofilm formation.
[00152] Thus the individual actives are most effective at inhibiting biofilm formation
at a w/w ratio of 0.0128%.
Example 3
Inhibition of biofilm formation by the combination of ascorbic acid and gallic
acid
[00153] The combination of gallic acid and ascorbic acid were then tested to
determine whether the composition would be effective in inhibiting biofilm formation.
For this purpose, a 1:1 ratio of gallic acid: ascorbic acid at varying weight/weight
concentrations was tested against the biofilm forming organism Streptococcus mutans.
[00154] Among the concentrations ranging from 0.0001%) to 0.0512%) of both gallic
acid and ascorbic acid, remarkably, both the actives, in combination, showed the
highest degree of percent inhibition at a concentration of 0.0128%) for each. The
percentage inhibition at this concentration was seen at 19.5% which was significantly
higher than the inhibition observed for the other ratios. Concentrations of both actives
below 0.0128%o showed very low inhibitory activity against biofilm formation, while
above this value, the concentrations demonstrated some inhibitory activity, however
this was not at levels comparable to when the actives were used at 0.0128%>.

[00155] Thus unexpectedly, at the concentration of 0.0128% each, the actives gallic acid and ascorbic acid in combination are most effective against biofilm formation as compared to the other concentrations. Example 3
Synergistic effect of the composition of ascorbic acid and gallic acid at specific w/w ratios
[00156] Since the 1:1 combination of 0.0128% (w/w) each of ascorbic acid and gallic acid exhibited the highest inhibition of biofilm formation, the efficacy of different combinations of the actives having a total effective concentration of 0.0256%) (w/w) was tested for activity against biofilm formation (Figure 3). Different combination of the actives covering the range of 1:4 to 4:1, ascorbic acid: gallic acid were tested for a total effective concentration of 0.0256%).
[00157]Unexpectedly, a ratio of 4:1 of ascorbic acid and gallic acid respectively, exhibited the highest ability to inhibit biofilm formation by S. mutans at 26.6 ± 3.5%. This was much higher than when 0.0256%) of the actives were used in isolation. Remarkably, as compared to a 1:1 ratio of ascorbic acid to gallic acid, the 4:1 ratio shows a significantly higher percentage inhibition when compared to the 1:1 w/w ratios at the different weight percentages.
[00158] Thus, a 4:1 ratio of the combination of ascorbic acid and gallic acid at a w/w percentage of 0.02048%) and 0.00512%), respectively, demonstrates remarkable synergism in the inhibition of biofilm formation.
[00159] Overall, the present disclosure provides a composition ascorbic acid and gallic acid, which exhibits a surprising and unexpected synergistic effect on bacterial biofilm formation. This apparent synergism of the said combination is not readily apparent at all concentration combinations (as discussed previously), suggesting that the results are not a result of mere admixture of the two or routine in nature. It is to be noted that it is within the purview of a person skilled in the art to incorporate the

disclosed composition into a host of formulations, particularly for the treatment of dental caries.

I/We Claim:
1. A composition comprising:
(a) ascorbic acid; and
(b) gallic acid,
wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:1.5-1:4.
2. The composition as claimed in claim 1, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:2.5-l :4.
3. The composition as claimed in claim 1, wherein gallic acid to ascorbic acid w/w ratio in said composition is 1:4.
4. A composition comprising:

(a) ascorbic acid having a w/w concentration in the range of 0.0064%-0.0512%, and
(b) gallic acid having a w/w concentration in the range of 0.0064%-0.0512%, wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:1.5-1:4.
5. A composition comprising:
(a) ascorbic acid having a w/w concentration in the range of 0.0128%; and
(b) gallic acid having a w/w concentration in the range of 0.0128%),
wherein gallic acid to ascorbic acid w/w ratio in said composition is in the range of 1:1.5-1:4.
6. The composition as claimed in claim 1-5, further comprising:
(a) at least one carrier;
(b) at least one diluent; and
(c) at least one excipient.
7. A method for preparing a composition as claimed in claim 1-5, said method
comprising the steps of:
(a) obtaining ascorbic acid;
(b) obtaining gallic acid; and

(c) contacting ascorbic acid and gallic acid to obtain said composition.
8. A method for preparing a composition as claimed in claim 6, said method
comprising the steps of:
(a) obtaining ascorbic acid;
(b) obtaining gallic acid;
(c) obtaining at least one carrier;
(d) obtaining at least one diluent;
(e) obtaining at least one excipient; and
(f) contacting ascorbic acid, gallic acid, at least one carrier, at least one diluent, and at least one excipient to obtain said composition.
9. A method for inhibiting bacterial biofilm formation, said method comprising:
(a) obtaining a composition as claimed in any of the claims 1-6;
(b) contacting said composition with bacteria capable of biofilm formation, wherein said composition inhibits bacterial film formation.
10. The method as claimed in claim 9, wherein the bacteria is Streptococcus mutans,