TECHNICAL FIELD
[001] The subject matter described herein in general relates to antimicrobial
compositions, and in particular, relates to the antibacterial compositions.
BACKGROUND OF INVENTION
[002] Antimicrobial compositions or antimicrobials have occupied a major space in the health care, food, cosmetic and household care sectors. For instance, antimicrobials can be used as food preservatives to curb the activities of harmful bacteria or find use in air purifiers to decrease the microbial load in the air. Conventionally used synthetic antimicrobials, such as triclosan, have been tightly regulated owing to its dangerous side-effects. There is an ever-increasing demand for antimicrobial agents, particularly anti-bacterial agents, due to increased antibiotic resistance.
[003] In the recent years, owing to environmental concerns, compositions derived from natural sources, are being increasingly used as desirable alternatives. WO2017083363 discloses the use of an antimicrobial composition that comprise a mixture of at least one Albizia extract and a Magnolia extract. WO2002022147 discloses a synergistic composition comprising an extract obtained from a neem plant, together with an effective amount of an antibiotic or chemotherapeutic anti-bacterial drug, the said composition being useful in the treatment of resistant bacterial infections. US6268399 discloses an antimicrobial composition comprising an effective amount of an antimicrobial agent comprising an extract of the plant Gaultheria shallon. However, these compositions have met with minimal acceptance in the marketplace owing to poor microbicidal effects and similar instances/symptoms of skin irritancy experienced as with synthetic chemical agents. [004] Considering the above evidence, there exists a need to develop a herbal composition that is devoid of synthetic ingredients, that is capable of imparting an antimicrobial effect.

SUMMARY OF THE INVENTION
[005] In an aspect of the present invention, there is provided a composition comprising: a) Acacia concinna extract; and b) citric acid, salts and derivatives thereof, wherein the Acacia concinna extract, and citric acid, salts and derivatives thereof, w/w ratio is in a range of 1:1 to 17:1.
[006] In another aspect of the present invention, there is provided a process for preparing the composition comprising: a) Acacia concinna extract; and b) citric acid, salts and derivatives thereof, wherein the Acacia concinna extract, and citric acid, salts and derivatives thereof, w/w ratio is in a range of 1:1 to 17:1, said process comprising: a) obtaining Acacia concinna extract; b) obtaining citric acid salts and derivatives thereof; and c) contacting the Acacia concinna extract with citric acid salts and derivatives thereof, to obtain the composition.
[007] 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.
DETAILED DESCRIPTION OF THE INVENTION
[008] 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
[009] 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.
[0010] KVS O`bWQZSa kOl* kO\l O\R kbVSl O`S caed to refer to one or to more than one
(i.e., to at least one) of the grammatical object of the article.
[0011] KVS bS`[a kQ][^`WaSl O\R kQ][^`WaW\Ul O`S caSR W\ bVS W\QZcaWdS* ]^S\ aS\aS*
meaning that additional elements may be included. It is not intended to be construed
Oa kQ]\aWaba ]T ]\Zgl,
[0012] Throughout this specification, unless the context requires otherwise the word
kQ][^`WaSl* O\R dO`WObW]\a acQV Oa kQ][^`WaSal O\R kQ][^`WaW\Ul* eWZZ PS
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.
[0013] KVS bS`[ kW\QZcRW\Ul Wa caSR b] [SO\ kW\QZcRW\U Pcb \]b ZW[WbSR b]l,
kC\QZcRW\Ul O\R kW\QZcRW\U Pcb \]b ZW[WbSR b]l O`S caSR W\bS`QVO\USOPZg,
[0014] 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. For example, a ratio range
of about 1:1 to 17:1 should be interpreted to include not only the explicitly recited
limits of about 1:1 to about 17:1, but also to include sub-ranges, such as 1:1, 4:1,
8:1,16:1 and so forth, as well as individual amounts, including fractional ratios,
within the specified ranges, such as 4.33:1, and 8.6:1, for example.
[0015] KVS bS`[ kOb ZSOab ]\Sl Wa caSR b] [SO\ ]\S ]` []`S O\R bVca W\QZcRSa
individual components as well as mixtures/combinations.
[0016] KVS bS`[ kSfb`OQbl `STS`a b] O preparation containing the active ingredient of
a substance in concentrated form. Exacts may be in the form of solids, powders, and
liquids.
[0017] ; Q][^]aWbW]\ Q][^`WaW\U kag\S`UWabWQ OQbWdWbgl ]` O kag\S`UWabWQ
Q][^]aWbW]\l Wa O Q][PW\ObW]\ ]T Q][^]c\Ra eVWQV SfVWPWba W\Q`SOaSR PW]Z]UWQOZ ]`
functional activity as a non-linear multiple of the biological or functional activity of

the individual compounds. In other words, the combined biological or functional activity of two or more compounds being tested is significantly greater than the expected result based on independent effects of the compounds when tested separately. Synergy may be apparent only at some ranges or weight percentages. [0018] The term derivatives refer to compounds that arise or be synthesized from a parent compound by replacement of one atom with another atom or group of atoms. [0019] Excipients are substances that serve as mechanisms to improve the delivery and the effectiveness of drugs
[0020] 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.
[0021] 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. [0022] As mentioned previously, conventionally used synthetic antimicrobials are toxic, not environmentally friendly; and are associated with side-effects such as, irritation on skin and mucous membrane of the user. Although herbal alternatives are being increasingly used as a desirable alternative, so far, these compositions have met with minimal acceptance in the marketplace owing to poor microbicidal effect and similar instances/symptoms of skin irritancy experienced as with synthetic chemical agents. The present disclosure aims to solve the said problem by using a combination of Acacia concinna extract and citric acid, which when used in defined w/w ratios is effective in providing a synergistic effect against microorganisms, particularly gram-negative bacteria.
[0023] In an embodiment of the present disclosure, there is provided a composition comprising: a) Acacia concinna extract; and b) citric acid, salts and derivatives

thereof, wherein the Acacia concinna extract, and citric acid, salts and derivatives
thereof, w/w ratio is in a range of 1:1 to 17:1.
[0024] In an embodiment of the present disclosure, there is provided a composition
comprising: a) Acacia concinna extract; and b) citric acid, wherein the Acacia
concinna extract, and citric acid w/w ratio is in a range of 1:1 to 17:1. In another
embodiment of the present disclosure, the Acacia concinna extract, and citric acid
w/w ratio is in a range of 4:1 to 17:1.
[0025] In an embodiment of the present disclosure, there is provided a composition
comprising: a) Acacia concinna extract; and b) citric acid, wherein the Acacia
concinna extract, and citric acid w/w ratio is 4.16:1.
[0026] In an embodiment of the present disclosure, there is provided a composition
comprising: a) Acacia concinna extract; and b) citric acid, wherein the Acacia
concinna extract, and citric acid w/w ratio is 8.33:1.
[0027] In an embodiment of the present disclosure, there is provided a composition
comprising: a) Acacia concinna extract; and b) citric acid, wherein the Acacia
concinna extract, and citric acid w/w ratio is 16.67:1.
[0028] In an embodiment of the present disclosure, there is provided a composition
as described herein, wherein the Acacia concinna extract has a weight percentage in
a range of 0.3 j 3% with respect to the extract; and wherein citric acid has a weight
percentage in a range of 0.03 - 0.4% with respect to the composition. In another
embodiment of the present disclosure, Acacia concinna extract has a weight
percentage in a range of 0.31 j 2.5% with respect to the extract; and wherein citric
acid has a weight percentage in a range of 0.03 - 0.3% with respect to the
composition.
[0029] In an embodiment of the present disclosure, there is provided a composition
comprising: a) Acacia concinna extract; and b) citric acid, wherein the Acacia
concinna extract, and citric acid w/w ratio is in a range of 1:1 - 17:1, and wherein
Acacia concinna extract has a weight percentage in a range of 0.3 j 3% with respect
to the extract; and wherein citric acid has a weight percentage in a range of 0.03 -
0.4% with respect to the composition.

[0030] In an embodiment of the present disclosure, there is provided a composition comprising: a) Acacia concinna extract; and b) citric acid, wherein the Acacia concinna extract, and citric acid w/w ratio is 4.16:1, and wherein Acacia concinna extract has a weight percentage in a range of 0.3 j 3% with respect to the extract; and wherein citric acid has a weight percentage in a range of 0.03 - 0.4% with respect to the composition.
[0031] In an embodiment of the present disclosure, there is provided a composition comprising: a) Acacia concinna extract; and b) citric acid, wherein the Acacia concinna extract, and citric acid w/w ratio is 8.33:1, and wherein Acacia concinna extract has a weight percentage in a range of 0.3 j 3% with respect to the extract; and wherein citric acid has a weight percentage in a range of 0.03 - 0.4% with respect to the composition.
[0032] In an embodiment of the present disclosure, there is provided a composition comprising: a) Acacia concinna extract; and b) citric acid, wherein the Acacia concinna extract, and citric acid w/w ratio is 16.67:1, and wherein Acacia concinna extract has a weight percentage in a range of 0.3 j 3% with respect to the extract; and wherein citric acid has a weight percentage in a range of 0.03 - 0.4% with respect to the composition.
[0033] In an embodiment of the present disclosure, there is provided a composition as described herein, wherein the Acacia concinna extract comprises saponin having a weight percentage in a range of 1 j 4% with respect to the extract. In another embodiment of the present disclosure, wherein the Acacia concinna extract comprises saponin having a weight percentage in a range of 2 j 2.5% with respect to the extract.
[0034] In an embodiment of the present disclosure, there is provided a composition comprising: a) Acacia concinna extract; and b) citric acid, wherein the Acacia concinna extract, and citric acid w/w ratio is in a range of 1:1 - 17:1, and wherein Acacia concinna extract comprises saponin having a weight percentage in a range of 1 j 4% with respect to the extract.
[0035] In an embodiment of the present disclosure, there is provided a composition comprising: a) Acacia concinna extract; and b) citric acid, wherein the Acacia

concinna extract, and citric acid w/w ratio is 4.16:1, and wherein Acacia concinna
extract comprises saponins having a weight percentage in a range of 1 j 4% with
respect to the extract.
[0036] In an embodiment of the present disclosure, there is provided a composition
comprising: a) Acacia concinna extract; and b) citric acid, wherein the Acacia
concinna extract, and citric acid w/w ratio is 8.33:1, and wherein Acacia concinna
extract comprises saponins having a weight percentage in a range of 1 j 4% with
respect to the extract.
[0037] In an embodiment of the present disclosure, there is provided a composition
comprising: a) Acacia concinna extract; and b) citric acid, wherein the Acacia
concinna extract, and citric acid w/w ratio is 16.67:1, and wherein Acacia concinna
extract comprises saponins having a weight percentage in a range of 1 j 4% with
respect to the extract.
[0038] In an embodiment of the present disclosure, there is provided a composition
comprising: a) Acacia concinna extract; and b) citric acid, wherein the Acacia
concinna extract, and citric acid w/w ratio is in a range of 1:1 - 17:1, and wherein
Acacia concinna extract has a weight percentage in a range of 0.3 j 3% with respect
to the extract; and wherein citric acid has a weight percentage in a range of 0.03 to
0.4% with respect to the composition, and wherein Acacia concinna extract
comprises saponins having a weight percentage in a range of 1 j 4% with respect to
the extract.
[0039] In an embodiment of the present disclosure, there is provided a composition
comprising: a) Acacia concinna extract; and b) citric acid, wherein the Acacia
concinna extract, and citric acid w/w ratio is in a range of 1:1 - 17:1, wherein Acacia
concinna extract has a weight percentage in a range of 0.31 j 2.5% with respect to
the extract; and wherein citric acid has a weight percentage in a range of 0.03 - 0.3%
with respect to the composition; and wherein the Acacia concinna extract comprises
saponins having a weight percentage in a range of 2 j 2.5% with respect to the
extract.
[0040] In an embodiment of the present disclosure, there is provided a composition
comprising: a) Acacia concinna extract; and b) citric acid, wherein the Acacia

concinna extract, and citric acid w/w ratio is 4.16:1, wherein Acacia concinna extract has a weight percentage in a range of 0.31 j 2.5% with respect to the extract; and wherein citric acid has a weight percentage in a range of 0.03 - 0.3% with respect to the composition; and wherein the Acacia concinna extract comprises saponins having a weight percentage in a range of 2 j 2.5% with respect to the extract. [0041] In an embodiment of the present disclosure, there is provided a composition comprising: a) Acacia concinna extract; and b) citric acid, wherein the Acacia concinna extract, and citric acid w/w ratio is 8.33:1, wherein Acacia concinna extract has a weight percentage in a range of 0.31 j 2.5% with respect to the extract; and wherein citric acid has a weight percentage in a range of 0.03 - 0.3% with respect to the composition; and wherein the Acacia concinna extract comprises saponins having a weight percentage in a range of 2 j 2.5% with respect to the extract. [0042] In an embodiment of the present disclosure, there is provided a composition comprising: a) Acacia concinna extract; and b) citric acid, wherein the Acacia concinna extract, and citric acid w/w ratio is 16.67:1, wherein Acacia concinna extract has a weight percentage in a range of 0.31 j 2.5% with respect to the extract; and wherein citric acid has a weight percentage in a range of 0.03 - 0.3% with respect to the composition; and wherein the Acacia concinna extract comprises saponins having a weight percentage in a range of 2 j 2.5% with respect to the extract. [0043] In an embodiment of the present disclosure, there is provided a process for preparing a composition comprising: a) Acacia concinna extract; and b) citric acid, salts and derivatives thereof, wherein the Acacia concinna extract, and citric acid, salts and derivatives thereof, w/w ratio is in a range of 1:1 to 17:1, said process comprising: a) obtaining the Acacia concinna extract; b) obtaining citric acid, salts and derivatives thereof; and c) contacting the Acacia concinna extract with citric acid, salts and derivatives thereof, to obtain the composition. [0044] In an embodiment of the present disclosure, there is provided a process for preparing a composition comprising: a) Acacia concinna extract; and b) citric acid; wherein the Acacia concinna extract, and citric acid, w/w ratio is in a range of 1:1 to 17:1, said process comprising: a) obtaining the Acacia concinna extract; b) obtaining

citric acid; and c) contacting the Acacia concinna extract with citric acid, to obtain the composition.
[0045] In an embodiment of the present disclosure, there is provided a process for preparing a composition comprising: a) Acacia concinna extract; and b) citric acid; wherein the Acacia concinna extract, and citric acid w/w ratio is in a range of 1:1 to 17:1, and wherein Acacia concinna extract has a weight percentage in a range of 0.3 j 3% with respect to the extract; and wherein citric acid has a weight percentage in a range of 0.03 to 0.4% with respect to the composition, said process comprising: a) obtaining the Acacia concinna extract; b) obtaining citric acid; and c) contacting the Acacia concinna extract with citric acid to obtain the composition. [0046] In an embodiment of the present disclosure, there is provided a process for preparing a composition comprising: a) Acacia concinna extract; and b) citric acid; wherein the Acacia concinna extract, and citric acid w/w ratio is in a range of 1:1 to 17:1, and wherein Acacia concinna extract has a weight percentage in a range of 0.3 j 3% with respect to the extract; and wherein citric acid has a weight percentage in a range of 0.03 to 0.4% with respect to the composition, and wherein the Acacia concinna extract comprises saponin having a weight percentage in a range of 1 j 4% with respect to the extract, said process comprising: a) obtaining the Acacia concinna extract; b) obtaining citric acid; and c) contacting the Acacia concinna extract with citric acid to obtain the composition.
[0047] In an embodiment of the present disclosure, there is provided a process for preparing a composition comprising: a) Acacia concinna extract; and b) citric acid; wherein the Acacia concinna extract, and citric acid w/w ratio is in a range of 1:1 to 17:1, and wherein Acacia concinna extract has a weight percentage in a range of 0.31 j 2.5% with respect to the extract; and wherein citric acid has a weight percentage in a range of 0.03 - 0.3% with respect to the composition, and wherein the Acacia concinna extract comprises saponin having a weight percentage in a range of 1 j 4% with respect to the extract, said process comprising: a) obtaining the Acacia concinna extract; b) obtaining citric acid; and c) contacting the Acacia concinna extract with citric acid to obtain the composition.

[0048] In an embodiment of the present disclosure, the composition as described herein is an antimicrobial composition. The applications include, personal care products such as shampoos, cosmetics, and soaps, and the like.
EXAMPLES
[0049] 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.
[0050] Conventionally used microbicidal compositions are antimicrobial that are toxic, not bio-safe, corrosive; and cause irritation on skin and mucous membrane of the user. Although essential oils obtained from natural sources is increasingly used as a desirable alternative, so far, these compositions have met with minimal acceptance in the marketplace owing to poor microbicidal effects and similar instances/symptoms of skin irritancy experienced as with synthetic chemical agents. The present disclosure aims to solve the said problem by using a combination of Acacia concinna extract and citric acid which when used in defined w/w ratios is effective in providing a synergistic effect against bacteria, particularly gram-negative bacteria.
Materials and methods
[0051] In order to test the antimicrobial activity of the actives, Acacia concinna powder was procured from Harika Chemicals (Product number-8906070780946); and anhydrous citric acid was procured from HiMedia (RM 1023-500G).
Example 1: Preparation of the inoculum
[0052] 10% Acacia concinna solution was prepared by dissolving 10g of Acacia
concinna powder in 100ml of water. A working stock solution of 5% Acacia

concinna powder (A) was prepared in Luria Bertani (LB) broth in a 96 well plate. The lower concentrations were obtained by 2-fold serial dilutions. These concentrations were tested in combination with citric acid (C) (0.15% and 0.075%). The combinations tested were A+C (0.625%+0.15%), A+C (1.25%+0.075%), A+C (0.625%+0.075%). E coli > DH5 alpha and S aureus j ATCC25923 inoculum of 106 CFU/mL density was added to the wells, such that the total volume in each well is 200uL. LB broth with the inoculum alone served as the growth control. Separate media blanks (without the inoculum) was maintained for each active and the combinations. The wells were incubated at 37°C with shaking for 18 hours and their optical density (OD) at every 10 minutes were read using a spectrophotometer. From the data collected the average lag phase, doubling time and the maximum OD relative to control was calculated; and the results are presented below from Table 1 j 4. [0053] Table 1 depicts the lag delay of E. coli in presence of Acacia concinna powder (A), and citric acid (C), alone and in combination. Lag delay is the period between the introduction of a microorganism into a culture medium and the time it begins to increase exponentially. A short lag phase is indicative of rapid multiplication of microorganisms.
[0054] As can be observed in Table 1, the average lag delay with Acacia concinna extract was found to be comparable with the control at both the tested concentrations; and similarly, the lag delay with citric acid was comparable with the control with 0.075% of citric acid. However, unexpectedly, and surprisingly, for the various combination ratios tested, it can be observed that the delay in lag phase in

comparison to the control for the combination ratios 4.16:1 and 16.67:1 was found to be 8.5 hours, 12 hours respectively. This delay in lag phase suggests that the rate of bacterial growth has been synergistically reduced with the composition of the present disclosure, particularly at w/w ratio of 16.67:1.
[0055] Table 2 depicts the lag delay of S. aureus in presence of Acacia concinna powder (A), and citric acid (C), alone and in combination.
[0056] As can be observed in Table 2, the average lag delay with Acacia concinna extract was found to be comparable with the control at both the tested concentrations, whereas with citric acid, the lag delay was found to be close to 15 hours. Also, the various combination ratios tested, it can be observed that the delay in lag phase was comparable to the citric acid. This suggests although the combination is effective against S. aureus, a synergistic effect in terms of bacterial (S. aureus) growth reduction was not observed with the composition of the present disclosure. [0057] Table 3 depicts a doubling time of E. coli in presence of Acacia concinna powder, and citric acid, alone and in combination. Doubling time is the time taken by the bacteria to double in number during a specified period. It varies with different organisms. For example, E. coli divides around every 20 minutes, hence its doubling time is 20 minutes, and for Staphylococcus aureus it is 30 minutes.

[0058] Although a delay in doubling time was observed in comparison to the control for the all concentrations tested, it can be observed that a doubling time was about 4 times slower than the control for the combination (A:C) ratio 4.16:1; and a complete inhibition in bacterial growth was observed for the combination (A:C) ratio 16.67:1. A higher average doubling time is indicative of slow multiplication, thereby suggesting that the composition of the present disclosure is effective in reducing the bacterial growth (E.coli).
[0059] Table 4 depicts a doubling time of Acacia concinna powder, and citric acid, alone and in combination against S. aureus.

[0060] As can be observed in Table 4, a complete growth inhibition was observed Ob OZZ Q]\QS\b`ObW]\a bSabSR eWbV QWb`WQ OQWR* O\R T]` OZZ Q][PW\ObW]\ `ObW]ma bSabSR, Further, the effect of the actives, Acacia concinna powder, and citric acid, alone and in combination at various w/w ratios was tested against E. coli, and the percentage inhibition in bacterial growth was calculated. The results are presented below in
[0061] For various combination ratios tested, it can be observed that a synergistic effect is observed for all the ratios ranging from 1:1 to 16.67:1. However, no synergistic effect was observed for the w/w ratios 0.05:1 and 33:3:1. The percentage inhibition exhibited by the individual actives (2.5% of the Acacia concinna extract and 0.075% of citric acid) was found to be 92% and 3% respectively; the additive effect being 95%. However. when a combination of actives (2.5% of A+0.075% of C) was used, the percentage inhibition of bacterial growth was found to be 102%, which is comparable to the additive effect observed with the individual concentration of actives. Similarly, no synergistic effect was observed for the w/w ratio 0.05:1 (0.015% of Acacia concinna extract, and 0.3% citric acid). The percentage inhibition exhibited with 0.3% citric acid alone is 100%. No increase in percentage inhibition was observed when a combination of actives (0.015% of Acacia concinna extract,

and 0.3% citric acid). The data suggests that the composition of the present disclosure in defined w/w ratios, i.e., from 1:1 to 17:1 is very effective inhibiting the growth of E. coli. The percentage inhibition exhibited by the combination of actives at these w/w ratios is far greater than the additive effect exhibited by the individual concentrations of actives.
[0062] Similarly, the effect of the actives, Acacia concinna powder, and citric acid, alone and in combination at various w/w ratios was tested against S. aureus, and the percentage inhibition in bacterial growth was calculated. The results are presented below in Table 6.
[0063] Although the composition of the present disclosure is effective against S. aureus, it can be observed that a synergistic inhibition was observed only for the combination ratio 4.16:1.
[0064] Overall, the data suggests that composition of the present disclosure shows an anti-bacterial activity against both E. coli (Gram negative) and S. aureus (Gram positive) microbes. However, a surprising antimicrobial synergy was observed only against E. coli at specific w/w ratios. No synergy was observed against S. aureus.
Advantages of the present disclosure
[0065] The present disclosure discloses a composition comprising herbal ingredients that is effective against gram-negative bacteria, particularly against E. coli. The composition comprising Acacia concinna extract; and citric acid in a range of 1:1 to 17:1, was found to synergistically inhibit or slow down the growth of bacteria, particularly gram-negative bacteria. Also, since the composition is completely

prepared from plant sources, there are no side-effects associated with the use of the composition. The combination was found to be more effective than commercially available synthetics.

I/We Claim:
1. A composition comprising:
a) Acacia concinna extract; and
b) citric acid, salts and derivatives thereof,
wherein the Acacia concinna extract, and citric acid, salts and derivatives thereof, w/w ratio is in a range of 1:1 to 17:1.
2. The composition as claimed in claim 1, wherein the Acacia concinna extract, and citric acid, salts and derivatives thereof, has a w/w ratio of 4.16:1.
3. The composition as claimed in claim 1, wherein the Acacia concinna extract, and citric acid, salts and derivatives thereof, has a w/w ratio of 8.33:1.
4. The composition as claimed in claim 1, wherein the Acacia concinna extract, and citric acid, salts and derivatives thereof, has a w/w ratio of 16.67:1
5. The composition as claimed in any of the claims 1-4, wherein the Acacia concinna extract has a weight percentage in a range of 0.3 j 3% with respect to the extract; and wherein citric acid, salts and derivatives thereof, has a weight percentage in a range of 0.03 to 0.4% with respect to the composition.
6. The composition as claimed in any one of the claims 1 to 5, wherein the Acacia concinna extract comprises saponins having a weight percentage in a range of 1 j 4% with respect to the extract.
7. A process for preparing the composition as claimed in claim 1, said process comprising:
a) obtaining the Acacia concinna extract;
b) obtaining citric acid, salts and derivatives thereof; and
c) contacting the Acacia concinna extract with citric acid, salts and derivatives thereof, to obtain the composition.
8. The composition as claimed in any one of the claims 1 to 6, wherein said composition inhibits growth of gram-negative bacteria.