TECHNICAL FIELD
[001] The subject matter described herein in general relates to the field of antimicrobial compositions, and in particular relates to antimicrobial compositions for use as air sanitizers and deodorizers.
BACKGROUND OF INVENTION
[002] People have concerns regarding the air we breathe owing to the increased microbial load in the indoor space. The spread of respiratory tract pathogens, such as pneumococcal bacteria and molds causing asthma / allergies, from people who are sick and carriers of the disease to others occupying the same space is a grave concern. Also, as urban residents are spending more of their time indoors, in offices and homes, reducing the microbial loads in the indoor air has become extremely important.
[003] Various compositions for suppressing, eliminating, or counteracting airborne microorganisms and malodors have been disclosed in the art. For example, EP3119405A1 discloses a sanitizing composition comprising: i) an alkoxy silyl ammonium film-forming compound; ii) a benzalkonium or benzethionium chloride, iii) a polymeric biguanide, and iv) a glycol carrier. US11765936 discloses a composition comprising triethylene glycol di- and/or mono-glycidyl ether as the malodor counteractant(s) for air sanitation and eliminating odor. Further, US7307053B2 discloses a formulation for sanitizing air, and treating odors comprising: a glycol selected from the group consisting triethylene glycol (TEG), dipropylene glycol, propylene glycol and mixtures thereof; a quaternary ammonium salt; and one or more corrosion inhibitors.
[004] Many of these antimicrobial compositions include synthetic, toxic and poisonous chemicals that can cause serious effects on human health and our environment. Further, with the use of synthetic chemicals, after extensive usage of these antimicrobial compositions, the microorganisms have been shown to build up a tolerance or resistance to some of these synthetic chemicals, which reduces the efficacy of these compositions. Moreover, synthetic chemicals can persist in the environment, which can be deleterious. Considering the above evidence, there

exists a need to develop non-toxic, environmentally friendly, microbial resistance-free antimicrobial compositions for effective use as air sanitizers and deodorizers.
SUMMARY OF THE INVENTION
[005] In an aspect of the present invention, there is provided an antimicrobial
composition comprising: a) lactic acid; and b) Sapindus mukorossi extract, wherein
lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to
1:4.
[006] In another aspect of the present invention, there is provided a process for
preparing the antimicrobial composition comprising: a) lactic acid; and b) Sapindus
mukorossi extract, wherein lactic acid to the Sapindus mukorossi extract weight
ratio is in a range of 1:0.125 to 1:4; said process comprising the steps of: a)
obtaining lactic acid; b) obtaining the Sapindus mukorossi extract; and c)
contacting lactic acid and the Sapindus mukorossi extract 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] 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.
[0011] 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”.
[0012] 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.
[0013] The term “including” is used to mean “including but not limited to”.
“Including” and “including but not limited to” are used interchangeably.
[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 range of 1:0.125 to 1:4, should be construed as including sub-ranges of
1:0.15 to 1:0.3, 1:1.5 to 1:3, 1:1.25 to 1:3.75, and so on.
[0015] The term “at least one” is used to mean one or more and thus includes
individual components as well as mixtures/combinations.
[0016] The term “extract” includes solids, such as powders, as well as liquid
substances extracted from a plant, containing its essence in a concentrated form.
[0017] The term “Minimum Inhibitory Concentration (MIC)” refers to the lowest
concentration of a chemical which prevents visible growth of a microorganism.

[0018] The term “Minimum Bactericidal Concentration (MBC)” refers to a
concentration resulting in microbial death as defined by the inability to re-culture
bacteria.
[0019] The term “air microbes” include both bacteria and fungi.
[0020] For the purposes of the present disclosure, the terms “fragrances”,
“fragrance solubilizers”, and “preservative” refer to the generally known
components used in the art. The specific components of the above-described
categories have been mentioned in the present disclosure wherever applicable.
Wherever not mentioned, it can be construed that any component falling under the
categories can be used as part of the composition.
[0021] 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.
[0022] 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.
[0023] Conventionally used synthetic antimicrobials like triclosan are associated
with dangerous side-effects and are tightly regulated. Further, with the use of
synthetic chemicals, the microorganisms have been shown to build up a tolerance
or resistance to some of these synthetics, which reduces the efficacy of these
compositions. Therefore, the resistance-free antimicrobial compositions from
natural plant sources, which are environmental friendly would be an alternative to
the current problem. Antimicrobials against which resistance has not been reported
include alcohols (ethanol, propanol) and organic acid (lactic acid, citric acid, sorbic
acid, acetic acid, propionic acid, salicylic acid). They act on bacterial cell
membrane and cause intercellular pH changes, thereby damaging the cell
membrane and mediating cell death. These chemicals are combined with natural

plant actives to obtain antimicrobial actives with good activity (WO2015/136366). The present disclosure describes an antimicrobial composition which is a combination of a fruit extract (Sapindus mukorossi) and an organic acid (lactic acid).
[0024] Escherichia coli (E. coli) are a large diverse group of bacteria found in the mammalian intestine. While they are generally safe, certain strains of E. coli have acquired plasmids that confer them virulence properties leading to diarrhoea and urinary tract infections amongst humans. They are present as food contaminants. Staphylococcus aureus (S. aureus) is a common human pathogen. It is found in the environment and on the human body, majorly the nasal cavity. It causes skin and soft tissue infections such as boils, furuncles and cellulitis. S. aureus has become a major bacterium of concern as it can build up in hospitals leading to higher incidences of hospital-acquired infections. Candida albicans (C. albicans) is a pathogenic yeast and used as a representative of fungi for lab studies.
[0025] The present disclosure, describes an anti-microbial composition which is a combination of a fruit extract and an organic acid (lactic acid).
[0026] Sapindus mukorossi, a member of the family Sapindaceae, is commonly known by several names such as soapnut, soapberry, washnut, reetha, aritha, dodan and doadni. It is a deciduous tree widely grown in upper reaches of Indo-Gangetic plains, Shivaliks and sub Himalayan tracts at altitudes from 200 m to 1500 m. The fruit is valued for the saponins (10.1%) present in the pericarp and constitutes up to 56.5% of the drupe known for inhibiting tumour cell growth. Recent literature suggests that S. mukorossi has antimicrobial, cytotoxic, insecticidal, fungicidal activities.
[0027] In an embodiment of the present disclosure, there is provided an antimicrobial composition comprising: a) lactic acid; and b) Sapindus mukorossi extract, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:4. In another embodiment of the present disclosure, the lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:2. [0028] In an embodiment of the present disclosure, there is provided an antimicrobial composition as described herein, wherein lactic acid has a weight

percentage in a range of 0.75 – 2.25% with respect to the composition. In another embodiment, the lactic acid has a weight percentage in a range of 1-2% with respect to the composition.
[0029] In an embodiment of the present disclosure, there is provided an antimicrobial composition comprising: a) lactic acid; and b) Sapindus mukorossi extract, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:4, and wherein lactic acid has a weight percentage in a range of 0.75 – 2.25% with respect to the composition.
[0030] In an embodiment of the present disclosure, there is provided an antimicrobial composition as described herein, wherein the Sapindus mukorossi extract has a weight percentage in range of 0.20 – 2.20% with respect to the composition. In another embodiment, the Sapindus mukorossi extract has a weight percentage in range of 0.50 – 2.0 % with respect to the composition.
[0031] In an embodiment of the present disclosure, there is provided an antimicrobial composition comprising: a) lactic acid; and b) Sapindus mukorossi extract, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:4, and wherein the Sapindus mukorossi extract has a weight percentage in range of 0.20 – 2.20% with respect to the composition.
[0032] In an embodiment of the present disclosure, there is provided an antimicrobial composition comprising: a) lactic acid; and b) Sapindus mukorossi extract, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:4, and wherein lactic acid has a weight percentage in a range of 0.75 – 2.25% with respect to the composition, and wherein the Sapindus mukorossi extract has a weight percentage in range of 0.20 – 2.20% with respect to the composition.
[0033] In an embodiment of the present disclosure, there is provided an antimicrobial composition as described herein, wherein the Sapindus mukorossi extract comprises saponins having a weight percentage in a range of 30 - 60% with respect to the extract. In another embodiment of the present disclosure, the Sapindus mukorossi extract comprises saponins having a weight percentage in a range of 35 – 55% with respect to the extract.

[0034] In an embodiment of the present disclosure, there is provided an antimicrobial composition comprising: a) lactic acid; and b) Sapindus mukorossi extract, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:4, and wherein the Sapindus mukorossi extract comprises saponins having a weight percentage in a range of 30-60% with respect to the extract.
[0035] In an embodiment of the present disclosure, there is provided an antimicrobial composition comprising: a) lactic acid; and b) Sapindus mukorossi extract, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:4, and wherein the Sapindus mukorossi extract has a weight percentage in range of 0.20 – 2.20% with respect to the composition, and wherein the Sapindus mukorossi extract comprises saponins having a weight percentage in a range of 30 - 60% with respect to the extract.
[0036] In an embodiment of the present disclosure, there is provided an antimicrobial composition comprising: a) lactic acid; and b) Sapindus mukorossi extract, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:4, and wherein lactic acid has a weight percentage in a range of 0.75 – 2.25% with respect to the composition, and wherein the Sapindus mukorossi extract has a weight percentage in range of 0.20 – 2.20% with respect to the composition, and wherein the Sapindus mukorossi extract comprises saponins having a weight percentage in a range of 30 - 60% with respect to the extract. [0037] In an embodiment of the present disclosure there is provided a composition as described herein, wherein the composition further comprises at least one excipient.
[0038] In an embodiment of the present disclosure, there is provided a formulation comprising: a) lactic acid; b) Sapindus mukorossi extract; and (c) at least one excipient, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:4.
[0039] In an embodiment of the present disclosure, there is provided a formulation comprising: a) lactic acid; b) Sapindus mukorossi extract; and (c) at least one excipient, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a

range of 1:0.125 to 1:4, and wherein lactic acid has a weight percentage in a range of 0.75 – 2.25% with respect to the formulation, and wherein the Sapindus mukorossi extract has a weight percentage in range of 0.20 – 2.20% with respect to the formulation.
[0040] In an embodiment of the present disclosure, there is provided a formulation comprising: a) lactic acid; b) Sapindus mukorossi extract; and c) at least one excipient, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:4, and wherein lactic acid has a weight percentage in a range of 0.75 – 2.25% with respect to the formulation, and wherein the Sapindus mukorossi extract has a weight percentage in range of 0.20 – 2.20% with respect to the formulation, and wherein the Sapindus mukorossi extract comprises saponins having a weight percentage in a range of 30 - 60% with respect to the extract. [0041] In an embodiment of the present disclosure, there is provided a formulation as described herein, wherein the at least one excipient is selected from a group consisting of fragrance, fragrance solubilizer, preservative, and combinations thereof. In another embodiment of the present disclosure, the fragrance is at least one selected from a group consisting of fruity (citrus), aqua, floral (lavendar, chamomile), and combinations thereof, the fragrance solubilizer is at least one selected from a group consisting of triton X-100, tween 20, tween 80, and combinations thereof, the preservative is at least one selected from a group consisting of phenoxyethanol, benzyl benzoate, methylisothiazolinone, zinc pyrithione, benzalkonium chloride, benzyl alcohol, grapefruit seed extract, and combinations thereof. In yet another embodiment of the present disclosure, and the preservative is phenoxyethanol, the fragrance solubilizer is triton X100. [0042] In an embodiment of the present disclosure, there is provided a formulation comprising: a) lactic acid; b) Sapindus mukorossi extract; and c) at least one excipient, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:4, and wherein the at least one excipient is selected from a group consisting of fragrance, fragrance solubilizer, preservative, and combinations thereof.

[0043] In an embodiment of the present disclosure, there is provided a formulation comprising: a) lactic acid; b) Sapindus mukorossi extract; and c) at least one excipient, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:4, and wherein lactic acid has a weight percentage in a range of 0.75 – 2.25% with respect to the formulation, and wherein the Sapindus mukorossi extract has a weight percentage in range of 0.20 – 2.20% with respect to the formulation, and wherein the at least one excipient is selected from a group consisting of fragrance, fragrance solubilizer, preservative, and combinations thereof, and wherein the fragrance is at least one selected from a group consisting of fruity (citrus), aqua, floral (lavendar, chamomile), and combinations thereof, and has a weight percentage in a range of 0.1- 1%, with respect to the formulation, and wherein the fragrance solubilizer is at least one selected from a group consisting of triton X-100, tween 20, tween 80, and combinations thereof , and has a weight percentage in a range of 0.2 – 0.5%, with respect to the formulation, and wherein the preservative is at least one selected from a group consisting of phenoxyethanol, benzyl benzoate, methylisothiazolinone, zinc pyrithione, benzalkonium chloride, benzyl alcohol, grapefruit seed extract, and combinations thereof. and has a weight percentage in a range of 0.2 – 0.5% with respect to the formulation.
[0044] In an embodiment of the present disclosure, there is provided a formulation comprising: a) lactic acid; b) Sapindus mukorossi extract; and c) at least one excipient, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:4, and wherein lactic acid has a weight percentage in a range of 0.75 – 2.25% with respect to the composition, and wherein the Sapindus mukorossi extract has a weight percentage in range of 0.20 – 2.20% with respect to the formulation, and wherein the Sapindus mukorossi extract comprises saponins having a weight percentage in a range of 30 -60% with respect to the extract, and wherein the at least one excipient is selected from a group consisting of fragrance, fragrance solubilizer, and preservative, and wherein the fragrance is at least one selected from a group consisting of fruity (citrus), aqua, floral (lavendar, chamomile), and combinations thereof, and has a weight percentage in a range of 0.1- 1.0% with respect to the formulation, and wherein the fragrance solubilizer is

at least one selected from a group consisting of triton X-100, tween 20, tween 80,
and combinations thereof , and has a weight percentage in the range of 0.2 – 0.5%,
with respect to the formulation, and wherein the preservative is at least one selected
from a group consisting of phenoxyethanol, benzyl benzoate,
methylisothiazolinone, zinc pyrithione, benzalkonium chloride, benzyl alcohol, grapefruit seed extract, and combinations thereof and has a weight percentage in the range of 0.2 – 0.5% with respect to the formulation.
[0045] In an embodiment of the present disclosure, there is present a formulation comprising: a) lactic acid; b) Sapindus mukorossi extract; and c) at least one excipient, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:2, and wherein lactic acid has a weight percentage in a range of 1 – 2% with respect to the formulation, and wherein the Sapindus mukorossi extract has a weight percentage in a range of 0.50 – 2.0 % with respect to the formulation, and wherein the at least one excipient is selected from a group consisting of fragrance, fragrance solubilizer, and preservative, and wherein the fragrance has a weight percentage in a range of 0.1 – 1.0% with respect to the composition, and wherein the fragrance solubilizer is triton X 100, and has a weight percentage in a range of 0.2- 0.5% with respect to the formulation, and wherein the preservative is phenoxyethanol, and has a weight percentage in a range of 0.2- 0.5% with respect to the formulation.
[0046] In an embodiment of the present disclosure, there is provided an antimicrobial composition as described herein, wherein said composition inhibits growth of gram-positive and gram-negative bacteria. In another embodiment of the present disclosure, the gram-positive bacterium is S. aureus, and the gram-negative bacterium is E. coli.
[0047] In an embodiment of the present disclosure, there is provided an antimicrobial composition comprising: a) lactic acid; and b) Sapindus mukorossi extract, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:4, and wherein said composition inhibits growth of gram-positive and Gram-negative bacteria.

[0048] In an embodiment of the present disclosure, there is provided an antimicrobial composition comprising: a) lactic acid; and b) Sapindus mukorossi extract, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:4, and wherein lactic acid has a weight percentage in a range of 0.75 – 2.25% with respect to the composition, and wherein the Sapindus mukorossi extract has a weight percentage in range of 0.2 – 2.2 % with respect to the composition, and wherein said composition inhibits growth of gram-positive and gram-negative bacteria.
[0049] In an embodiment of the present disclosure, there is provided an antimicrobial composition comprising: a) lactic acid; and b) Sapindus mukorossi extract, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:4, and wherein lactic acid has a weight percentage in a range of 0.90 – 2.1% with respect to the composition, and wherein the Sapindus mukorossi extract has a weight percentage in range of 0.4 – 2.1 % with respect to the composition, and wherein said composition inhibits growth of gram-positive and gram-negative bacteria.
[0050] In an embodiment of the present disclosure, there is provided an antimicrobial composition comprising: a) lactic acid; and b) Sapindus mukorossi extract, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:4, and wherein lactic acid has a weight percentage in a range of 0.75 – 2.25% with respect to the composition, and wherein the Sapindus mukorossi extract has a weight percentage in range of 0.20 – 2.2 % with respect to the composition, and wherein the Sapindus mukorossi extract comprises saponins having a weight percentage in a range of 30 - 60% with respect to the extract, and wherein said composition inhibits growth of gram-positive and gram-negative bacteria.
[0051] In an embodiment of the present disclosure, there is provided an antimicrobial composition comprising: a) lactic acid; and b) Sapindus mukorossi extract, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:4, and wherein the composition further comprises at least one excipient selected from a group consisting of fragrance, fragrance solubilizer,

preservative and combinations thereof, and wherein said composition inhibits growth of gram-positive and gram-negative bacteria
[0052] In an embodiment of the present disclosure, there is provided an antimicrobial composition as described herein, wherein said composition inhibits growth of fungi. In another embodiment of the present disclosure, the fungus is C. albicans.
[0053] In an embodiment of the present disclosure, there is provided an antimicrobial composition comprising: a) lactic acid; and b) Sapindus mukorossi extract, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:4, and wherein said composition inhibits growth of fungi. [0054] In an embodiment of the present disclosure, there is provided an antimicrobial composition comprising: a) lactic acid; and b) Sapindus mukorossi extract, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:4, and wherein lactic acid has a weight percentage in a range of 0.75 – 2.25% with respect to the composition, and wherein the Sapindus mukorossi extract has a weight percentage in range of 0.20 – 2.2 % with respect to the composition, and wherein said composition inhibits growth of fungi. [0055] In an embodiment of the present disclosure, there is provided an antimicrobial composition comprising: a) lactic acid; and b) Sapindus mukorossi extract, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:4, and wherein lactic acid has a weight percentage in a range of 0.75 – 2.25% with respect to the composition, and wherein the Sapindus mukorossi extract has a weight percentage in range of 0.50 – 2.0 % with respect to the composition, and wherein the Sapindus mukorossi extract comprises saponins having a weight percentage in a range of 30-60% with respect to the extract, and wherein said composition inhibits growth of fungi.
[0056] In an embodiment of the present disclosure, there is provided an antimicrobial composition comprising: a) lactic acid; and b) Sapindus mukorossi extract, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:4, and wherein the composition further comprises at least one excipient selected from a group consisting of fragrance, fragrance solubilizer,

preservative and combinations thereof, and wherein said composition inhibits growth of fungi.
[0057] In an embodiment of the present disclosure, there is provided an antimicrobial composition as described herein, wherein the composition inhibits growth of bacteria and fungi. In another embodiment of the present disclosure, the bacteria comprise gram-positive bacteria and gram-negative bacteria, and fungi is C. albicans.
[0058] In an embodiment of the present disclosure, there is provided an antimicrobial composition as described herein, wherein the composition inhibits growth of bacteria and fungi in a mixed culture.
[0059] In an embodiment of the present disclosure, there is provided a formulation as described herein, wherein the formulation inhibits growth of bacteria and fungi. In another embodiment of the present disclosure, the bacteria comprise gram-positive bacteria and gram-negative bacteria, and fungi is C. albicans.
[0060] In an embodiment of the present disclosure, there is provided a formulation as described herein, wherein the composition inhibits growth of bacteria and fungi in a mixed culture.
[0061] In an embodiment, there is provided a process for preparing an
antimicrobial composition comprising: a) lactic acid; and b) Sapindus mukorossi extract, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:4, said process comprising the steps of: a) obtaining lactic acid; b) obtaining the Sapindus mukorossi extract; and c) contacting lactic acid and the Sapindus mukorossi extract to obtain the composition.
[0062] In another embodiment of the present disclosure, there is provided a process for preparing a n antimicrobial composition comprising: a) lactic acid; and b) Sapindus mukorossi extract, wherein lactic acid, and the Sapindus mukorossi extract w/w ratio is in a range of 1:0.125 to 1:4, and wherein lactic acid has a weight percentage in a range of 0.75 – 2.25% with respect to the composition, and wherein the Sapindus mukorossi extract has a weight percentage in a range of 0.20 – 2.2 % with respect to the composition, and wherein said process comprising the steps of: a) obtaining lactic acid; b) obtaining the Sapindus mukorossi extract; and

c) contacting lactic acid and the Sapindus mukorossi extract to obtain the composition.
[0063] In another embodiment of the present disclosure, there is provided a process for preparing a composition comprising: a) lactic acid; and b) Sapindus mukorossi extract, wherein lactic acid, and the Sapindus mukorossi extract w/w ratio is in a range of 1:0.125 to 1:4, and wherein lactic acid has a weight percentage in a range of 0.75 – 2.25% with respect to the composition, and wherein the Sapindus mukorossi extract has a weight percentage in a range of 0.20 – 2.2 % with respect to the composition, and wherein the Sapindus mukorossi extract comprises saponins having a weight percentage in a range of 30 - 60% with respect to the extract, and wherein said process comprising the steps of: a) obtaining lactic acid; b) obtaining the Sapindus mukorossi extract; and c) contacting lactic acid and the Sapindus mukorossi extract to obtain the composition.
[0064] In another embodiment of the present disclosure, there is provided a process for preparing a formulation comprising: a) lactic acid; b) Sapindus mukorossi extract; and (c) at least one excipient, wherein lactic acid, and the Sapindus mukorossi extract w/w ratio is in a range of 1:0.125 to 1:4, and wherein the at least one excipient is selected from a group consisting of fragrance, fragrance solubilizer, preservatives, and combinations thereof, said process comprising the steps of: a) obtaining lactic acid; b) obtaining the Sapindus mukorossi extract; c) obtaining the at least one excipient; and d) contacting lactic acid, the Sapindus mukorossi extract, with the at least one excipient, to obtain the formulation. [0065] In another embodiment of the present disclosure, there is provided a process for preparing a formulation comprising: a) lactic acid; b) Sapindus mukorossi extract; and c) at least one excipient, wherein lactic acid, and the Sapindus mukorossi extract w/w ratio is in the range of 1:0.125 to 1:4, and wherein lactic acid has a weight percentage in a range of 0.75 – 2.25% with respect to the composition, and wherein the Sapindus mukorossi extract has a weight percentage in range of 0.20 – 2.2 % with respect to the composition, and wherein the at least one excipient is selected from a group consisting of fragrance, fragrance solubilizer, preservatives, and combinations thereof, and wherein said process

comprising the steps of: a) obtaining lactic acid; b) obtaining the Sapindus mukorossi extract; c) obtaining the at least one excipient; and d) contacting lactic acid, the Sapindus mukorossi extract, with the at least one excipient, to obtain the formulation.
[0066] In another embodiment of the present disclosure, there is provided a process for preparing a formulation comprising: a) lactic acid; and b) Sapindus mukorossi extract, and c) at least one excipient, wherein lactic acid, and the Sapindus mukorossi extract w/w ratio is in a range of 1:0.125 to 1:4, and wherein lactic acid has a weight percentage in a range of 0.75 – 2.25% with respect to the composition, and wherein the Sapindus mukorossi extract has a weight percentage in a range of 0.20 – 2.2 % with respect to the formulation, and wherein the Sapindus mukorossi extract comprises saponins having a weight percentage in a range of 30 -60% with respect to the extract, and wherein the at least one excipient is selected from a group consisting of fragrance, fragrance solubilizer, preservatives, and combinations thereof, said process comprising the steps of: a) obtaining lactic acid; b) obtaining the Sapindus mukorossi extract; c) obtaining the at least one excipient; and d) contacting lactic acid, the Sapindus mukorossi extract, with the at least one excipient, to obtain the formulation.
[0067] In an embodiment of the present disclosure, there is provided a formulation as described herein, wherein the formulation is delivered using an ultrasonic humidifier device at a frequency of 0.8 to 3 MHz for a period of 1-2 h.
[0068] In an embodiment of the present disclosure, there is present a formulation comprising: a) lactic acid; b) Sapindus mukorossi extract; and c) at least one excipient, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:4, and wherein the at least one excipient is selected from a group consisting of fragrance, fragrance solubilizer, preservatives, and combinations thereof wherein the formulation is delivered using a ultrasonic humidifier device at a frequency of 0.8 to 3 MHz for a period of 1-2 h.
[0069] In an embodiment, the antimicrobial composition of the present disclosure can be used as a food preservative.

[0070] In another embodiment, the antimicrobial composition of the present disclosure can be used as air sanitizer and deodorizer to decrease microbial load in the air.
[0071] 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
[0072] 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. [0073] The working examples described herein clearly depict the effect of the combination of organic acid, and a fruit extract in defined w/w ratios for use in inhibiting bacterial and fungal growth. The present disclosure is directed to an antimicrobial composition comprising: a) lactic acid; and b) Sapindus mukorossi extract, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:4, for use as an air sanitizer and deodorizer to decrease the microbial load in the air. Further, the antimicrobial composition can be delivered with cold mist generated upon using the ultrasonic humidifier, when operated at a frequency of 0.8 to 3 MHz, for a period of 1-2h. This device generates mists of the

composition which spreads in to the room giving better spread and delivery of the composition.
MATERIALS AND METHODS
Example 1: Evaluation of antimicrobial activity of the actives
[0074] Actives evaluated for antimicrobial activity: Momordica charantia (product
no. MC/09010), Mangifera indica (product no. RD/2285), Phyllanthus amarus
(product no. RD/2237), Punica granatum (product no. PG/10003), Syzygium
cumini (product no. SC/09002), Terminalia chebula (product no. TA/09003) were
procured from Natural Remedies, Bangalore. Sundried and seedless Sapindus
mukorossi (Aritha) powder was obtained from Prakruti Products Pvt. Ltd., Karwar
(Product number-SM-17114). Actives were dissolved in dimethyl sulfoxide
(DMSO) at concentration of 20 mg/ml. Lactic acid was procured from Fluka
(Catalog number-69775).
[0075] Cultivation and Preparation of microbial cultures: Escherichia coli
DH5alpha and Staphylococcus aureus ATCC 25923 were used as representative of
gram-positive and gram-negative bacteria, respectively. Candida albicans, MTCC
227 served as a fungal representative.
[0076] They were inoculated in 4 ml sterile Luria Bertani Broth (LB) broth
(HiMedia, Mumbai; Cat no. M1245) in a 15 ml sterile tubes and cultivated for 24h
at 37°C. Overnight culture were grown in fresh LB broth and grown for 2-3h to
obtain mid-log phase cells. After cultivation, the cells were harvested by
centrifugation (2 min, 5000 x g). The cell pellet was washed twice in fresh LB
broth. Aliquot was diluted to an optical density OD600nm of 0.1 and used for further
experiments.
[0077] Testing of antimicrobial activity of actives: Antimicrobial activities of
Momordica charantia, Mangifera indica, Phyllanthus amarus, Punica granatum,
Syzygium cumini, Terminalia chebula, and Sapindus mukorossi were evaluated for
their activity against E. coli, S. aureus and C. albicans cultures, which were
prepared as mentioned above. The assay was carried out in 48 well plates. LB
broth (245 µl) was added in to wells. The stocks of actives prepared in DMSO (5
µl) were added into LB broth in separate wells in duplicate. The cultures adjusted

to O.D.600 nm of 0.1 were further diluted 1:100 in LB broth. 250 µl of this diluted culture was added to the wells containing LB broth with (test) and without (growth control) chemicals being tested. The 48 well plate was incubated at 37°C for 24 h. The growth or absence of growth of microbial cells was determined by visual observations for turbidity in the growth medium. Presence of small amount of growth / turbidity was considered as growth observed (+). Absence of turbidity was considered as “growth inhibited”. The results obtained are represented in Table 1. Table 1: Antimicrobial activities of natural extracts against Candida albicans
Microorganism tested for growth
Extract Concentration C. albicans E. coli S. aureus
(ppm)
Sapindus mukorossi 1000 No Growth + +
Momordica charantia 1000 + + +
Mangifera indica 1000 + + +
Phyllanthus amarus 1000 + + +
Punica granatum 1000 + + +
Syzygium cumini 1000 + + +
Terminalia chebula 1000 + + +
‘+’ is indicative of growth observed
From the Table 1, it can be observed that Sapindus mukorossi contains antimicrobial activity against Candida albicans, a fungus, but not against bacteria (E. coli and S. aureus). None of the other extracts exhibited antimicrobial activity against bacteria or fungi.
Example 2: Determination of Minimum Inhibitory Concentration (MIC) [0078] MIC of lactic acid against S. aureus, E. coli and C. albicans was determined for evaluating its antibacterial activity. MIC values were determined using 48 well plate (Corning, USA). Stock solution of lactic acid (10%) was prepared in LB broth. A 2-fold serial-dilution in LB broth (volume 250 µl) was done using this stock as the first concentration to obtain six concentrations in the

range of 5% - 0.156%. This was achieved by adding 250 µl of active containing LB broth in to 250 µl of plain LB broth to obtain 2-fold dilution, and sequential performing the same with the resultant new solution containing the active. Log phase cultures of E. coli, S. aureus and C. albicans, adjusted to O.D.600 nm of 0.1 were diluted 1:100 in LB broth and 250 µl aliquots of these were added to the wells containing various concentration of lactic acid containing LB broth. This resulted in final lactic acid concentrations of 2.5%, 1.25%, 0.625%, 0.313%, 0.156% and 078% lactic acid. The 48 well plate was incubated at 37°C for 24 h. The growth or absence of growth of bacterial cells was measured by visual observations for turbidity in the growth medium. The lowest concentration that showed no visible growth indicates the MIC. The results obtained are represented in Table 2. Table 2
Growth of Microorganisms
Concentration C. albicans E. coli S. aureus
2.5% + No Growth No Growth
1.25% + No Growth No Growth
0.625% + No Growth No Growth
0.313% + + +
0.156% + + +
0.078% + + +
0% (Control) + + +
‘+’ is indicative of growth observed
From the Table 2, it can be observed that lactic acid has ability inhibit growth of bacteria (E. coli and S. aureus). The minimum concentrations of lactic acid required to inhibit the growth was both E. coli and S. aureus was found to be 0.625%. It has no antimicrobial activity against Candida albicans, a fungus, in the concentration ranges tested.
Example 3: Determination of Minimum Bactericidal Concentration (MBC) and Minimum Fungicidal Concentration (MFC)

[0079] While MIC is the minimum concentration required for growth inhibitory activity of antibacterial substances, MBC and MFC is the minimum concentration required for its killing activity against bacteria and fungus, respectively. The experiment was done by challenging E. coli or S aureus at cell density of 1 million cells/ml against lactic acid at concentrations 4%, 2%, 1% and 0.5%, and additional 0.625% lactic acid concentration (MIC values) in LB broth. LB broth without lactic acid was used as growth control. After 24 h incubation at 37°C, 10 µl of the inoculated media were spotted on surface of sterile LB agar (HiMedia Laboratories, India) in 90 mm petridish using aseptic techniques. The petridish was incubated at 37°C for 24 h. The spots were evaluated for presence or absence of bacterial colonies formed from surviving bacteria in the 10 µl spotted volume. Absence of any colonies indicates at least 3 log kills required to be counted as bactericidal activity. The lowest concentration that showed no colonies on Brain Heart Fusion (BHI) agar plate indicates the MBC.
[0080] Similarly, MFC concentrations of Sapindus mukorossi was determined against C. albicans. 10% Sapindus mukorossi (Aritha) powder solution was prepared by adding 25 g of powder to 250 ml water. It was heated on hot water plate to boiling, cooled and then filtered through 0.2 um filter to sterilize and remove particulate matter. The experiment was further done by challenging C. albicans at cell density of 105 cells/ml against lactic acid at concentrations 8%, 4%, 2%, 1%, 0.5% 0.25%, 0.125%, 0.0625%, 0.032% and additional 0.625% Sapindus mukorossi extract concentration (MIC values) in LB broth. LB broth without Sapindus mukorossi extract was used as growth control. After 24 h incubation at 37°C, 10 µl of the inoculated media were spotted on surface of sterile LB agar (HiMedia Laboratories, India) in 90 mm petridish using aseptic techniques. The petridish was incubated at 37°C for 24 h. The spots were evaluated for presence or absence of fungal colonies formed from surviving fungus in the 10 µl spotted volume. Absence of any colonies indicates at least 2 log kills required to be counted as fungicidal activity. The lowest concentration that showed no colonies on SDA agar plate indicates the MFC. The results are presented in Table 3 (MBC), and Table 4 (MFC).

Table 3
Bacterial colonies
Concentration E. coli S. aureus
8% No Colonies No Colonies
4% No Colonies No Colonies
2% No Colonies No Colonies
1% No Colonies No Colonies
0.625% + +
0.5% + +
0% (Control) + +
MFC 1% 1%
‘+’- Bacterial colonies observed in spotted samples on LB agar
Table 4
Concentration Colonies of C. albicans
8.00%
No Colonies
4.00%
No Colonies
2.00%
No Colonies
1%
+
0.50%
+
0.25%
+
0.125%
+
0.063%
+
0% (Control) +
MFC 2%
‘+’: Fungal colonies observed in spotted samples on Sabouraud Dextrose Agar
(SDA).
[0081] As represented in Table 3, the minimum bactericidal concentrations of
lactic acid required to kill E. coli and S. aureus is 1%. As reported in Table 4, the

minimum fungicidal concentration of Sapindus mukorossi required to kill C. albicans is 2%. Hence, these concentrations were further evaluated for their efficacies in in situ conditions.
Example 4: Evaluation of Sapindus mukorossi extract and lactic acid to kill microorganisms in air
[0082] Evaluation study was done in windowless room of 200 sq. feet area with its door kept closed during the course of study. The sampling of air was done using air sampler (Model LA637) from HiMedia using a flow rate of 100 liters of air / minute for 3 minutes. An air sanitization solution consisting of 2% Sapindus mukorossi and 1% lactic acid solution was dispersed in room using an ultrasound based portable room humidifier (Model DT-1508B Allin Exporters). Alternatively, a mist spray bottle was used to spray 18 ml of the solution in to the room. Control experiment was done by applications of plain water instead of air sanitation solution. The microbial counts were taken before application of the air sanitization solution and 20 minutes after application of air sanitization solution. The bacterial counts were determined using agar plates of Trypticase Soy Agar (TSA) with lecithin and tween 80 supplemented with penicillin-streptomycin. The fungal counts were determined using agar plates made with Sabouraud Dextrose Agar (SDA) supplemented with cyclohexamide. The plates were incubated at 30°C for 4 days for development of bacterial and fungal colonies. Table 5
Counts (cfu/m3 of air) Inhibition (%)
Application Bacteria Fungi Bacteria Fungi
type Sample Before After Before After
Mist spray Control 343.33 326.67 193.33 170.00 4.9 12.1
Test 403.33 373.33 203.33 183.33 7.4 9.8
Humidifier Control 480.00 243.33 226.67 103.33 49.3 54.4
Test 356.67 60.00 236.67 36.67 83.2 84.5
[0083] The data in Table 5 shows that microbial count reductions are observed with the application of air sanitizing solution using humidifier. From the Table 5, it

can be inferred that the percentage reduction in air microbes was found to be in the range of 49 - 85% on use of the antimicrobial composition through the air humidifier. The percentage reduction in air microbes was found to be marginal, i.e., in the range of 4 - 12%, in comparison to the control set of experiment when the same solution was applied using a mist spray mode of application. The humidifier thus seems to allow for better dispersal of air sanitizing solution, allowing for greater impact in terms of air sanitation. The sampling solution is effective in reducing both bacterial and fungal counts in the air. Considering that air sampling requires an entry of a person in the room to operate the instrument, it is expected that the human activity inside the room will add on to the microbial loads in the room. Hence, it is not possible to get 100% microbial reduction without rapid processes.
Example 5: Evaluation of concentrations of Sapindus mukorossi extract and lactic acid for air sanitation
[0084] The air microbes were collected by air sampler in 10 ml LB broth placed in petriplate in an air sampler over a period of 10 minutes. It was used as air microbial mix culture. 250 ul of the mix culture was inoculated in 250 ul of various combination of lactic acid and Sapindus mukorossi extract concentrations prepared in water and was set up in a 48 well plate. After 1 h of contact time, 30 ul of each treatment was spotted on to TSA with lecithin and tween 80 plate to determine survival. The plates were incubated at 30°C for 1 day and observed for growth in spots. Any growth appeared was considered due to survival of cells during the treatment, and hence indicated lack of killing activity. No growth in the spots were considered as effective killing of air microbes. The results are presented in Table 6. Table 6
S. mukorossi I Lactic acid concentration
extract i 2% I 1% 0.50% 0.25% 0.125% 0%
4.00% - - - - - -
2.00% + + - - - -

1.00% + + - - - -
0.50% + + - - - -
0.25% +
0.00%
Key: – No Killing activity, + killing activity
[0085] The data in Table 6 indicates that the air microbes are effectively killed by concentration above 1% lactic acid and at 0.5% to 2% S. mukorossi extract. S. mukorossi extract at higher concentrations is not effective, perhaps due to protective activity from nutrients / phytochemicals from S. mukorossi extract. Based on these results as observed in Table 1 - 6, a prototype formulation for air sanitation is provided in Table 7. Table 7
S. No Ingredient Concentration Activity
1 S. mukorossi 0.5% - 2% Antimicrobial extract
2 Lactic acid 1% - 2% Antimicrobial
3 Fragrance as required Perfume
4 Triton X-100 0.5% Fragrance solubilizer
5 Phenoxyethanol 0.4% Preservative
6 Water to volume Solvent
The formulation as provided in Table 7 was prepared by mixing S. mukorossi extract, lactic acid, fragrance, triton X-100, phenoxyethanol, and water at a temperature range of 25 - 37℃ to obtain the formulation.
Advantages of the present disclosure:
[0086] The present disclosure discloses an antimicrobial composition comprising: a) lactic acid; and b) Sapindus mukorossi extract, wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:4. The present disclosure reveals that the compositions of the present disclosure are effective for

use in air sanitation and can impart deodorizing properties. Further, the composition when delivered through an ultrasound humidifier is effective in decreasing the microbial load in the air. The use of this combination in an air humidifier circumvents the need for use of expensive air purifiers and air conditions for decreasing the air microbes. Furthermore, the compositions of the present disclosure are non-toxic, environmentally friendly, cost-effective, and are effective against strains of antimicrobial resistant microbes.

I/We Claim:
1. An antimicrobial composition comprising:
a) lactic acid; and
b) Sapindus mukorossi extract,
wherein lactic acid to the Sapindus mukorossi extract weight ratio is in a range of 1:0.125 to 1:4.
2. The antimicrobial composition as claimed in claim 1, wherein lactic acid has a weight percentage in a range of 0.75 – 2.25% with respect to the composition.
3. The antimicrobial composition as claimed in claim 1, wherein the Sapindus mukorossi extract has a weight percentage in range of 0.20 – 2.20% with respect to the composition.
4. The antimicrobial composition as claimed in claim 1, wherein the Sapindus mukorossi extract comprises saponins having a weight percentage in a range of 30 - 60% with respect to the extract.
5. A formulation comprising:

a) the composition as claimed in any one of the claims 1-4; and
b) at least one excipient.

6. The formulation as claimed in claim 5, wherein the at least one excipient is selected from a group consisting of fragrance, fragrance solubilizers, preservatives, and combinations thereof.
7. The antimicrobial composition as claimed in any one of the claims 1-4, wherein said composition inhibits growth of Gram-positive and Gram-negative bacteria.
8. The antimicrobial composition as claimed in any one of the claims 1-4, wherein said composition inhibits growth of fungi.
9. A process for preparing the composition as claimed in claim 1, said process comprising the steps of: a) obtaining lactic acid; b) obtaining Sapindus mukorossi extract; and c) contacting lactic acid with the Sapindus mukorossi extract to obtain the composition.

10. A process for preparing the formulation as claimed in claim 5, said process comprising the steps of: a) obtaining lactic acid; b) obtaining Sapindus mukorossi extract; and c) contacting lactic acid and the Sapindus mukorossi extract with the at least one excipient to obtain the formulation.
11. The formulation as claimed in claim 5, wherein the formulation is delivered using an ultrasonic humidifier device for a period of 1 – 2 h at a frequency of 0.8 – 3 MHz.