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Essential Oil Compositions From Pancha Tulasi Drops

Patent Information

Application #
Patent Number
Invention Field
PHARMACEUTICALS
Publication Type
INA
Publication Number
08/2015
Status
Legal Status
Filing Date
27 January 2015
Grant Date
2022-05-06
Renewal Date

Abstract

Abstract: Composition for disinfecting water contaminated with pathogenic microorganisms, . comprising: a) an emulsifier; b) an essential oil composition selected from the group consisting of basil oils of Ocimum sanctum; Ocimum basilicum; Ocimum grattissimum, Ocimum canum, Ocimum citriodorum, Ocimum tenuiflorum L, Ocimum ciliatum, Ocimum kilimandscharicum, Ocimum americanum; Ocimum campechianum, Ocimum Viride ; c) optionally a carrier oil comprising a plant oil; d) optionally a natural anti-oxidant present in basil preserving the shelf life as well as provide anti-oxidant effect; _ e) optionally containing an ingredient(s) enhances natural immunity. The present invention also provides a method for disinfecting water contaminated with pathogenic microorganisms, comprising treating the contaminated potable drinking water instantly with an effective amount of the composition.

Applicants

1. BABUULAL BHAVARLAL JAIN
555 GREEN PARK APARTMENT, DR. PRADHAN MARG, CHARAI, THANE (WEST), 400601, MAHARASHTRA, INDIA
2. SURESHKUMAR BABUULAL JAIN
555 GREEN PARK APARTMENT, DR. PRADHAN MARG, CHARAI, THANE (WEST), 400601, MAHARASHTRA, INDIA

Inventors

1. SURESHKUMAR BABUULAL JAIN
555 GREEN PARK APARTMENT, DR. PRADHAN MARG, CHARAI, THANE (WEST), 400601, MAHARASHTRA, INDIA
2. AASINKUMAR JAGNARAYAN MAURYA
12 SINDH SAGAR, PLOT 92, OPP. ANDHRA SCHOOL, WADALA (WEST), MUMBAI-400031, MAHARASHTRA, INDIA

Eregister



Year CBR Date CBR Number Renwal Amount Renwal Date Normal Due Date Renwal To Renwal From Due Date with Extension Reneal Certificate Number
3rd year 24/09/2024 46841 800 24/09/2024 06/08/2022 27/01/2018 27/01/2017 06/02/2023 85555
4th year 24/09/2024 46841 800 24/09/2024 06/08/2022 27/01/2019 27/01/2018 06/02/2023 85556
5th year 24/09/2024 46841 800 24/09/2024 06/08/2022 27/01/2020 27/01/2019 06/02/2023 85557
6th year 24/09/2024 46841 800 24/09/2024 06/08/2022 27/01/2021 27/01/2020 06/02/2023 85558
7th year 24/09/2024 46841 2400 24/09/2024 06/08/2022 27/01/2022 27/01/2021 06/02/2023 85559
8th year 24/09/2024 46841 2400 24/09/2024 06/08/2022 27/01/2023 27/01/2022 06/02/2023 85560
9th year 24/09/2024 46841 2400 24/09/2024 27/01/2023 27/01/2024 27/01/2023 27/07/2023 85561
10th year 24/09/2024 46841 2400 24/09/2024 27/01/2024 27/01/2025 27/01/2024 27/07/2024 85562
11th year 28/10/2024 50989 4800 28/10/2024 27/01/2025 27/01/2026 27/01/2025 27/07/2025 90730
12th year 28/10/2024 50989 4320 28/10/2024 27/01/2026 27/01/2027 27/01/2026 27/07/2026 90731
13th year 28/10/2024 50989 4320 28/10/2024 27/01/2027 27/01/2028 27/01/2027 27/07/2027 90732
14th year 28/10/2024 50989 4320 28/10/2024 27/01/2028 27/01/2029 27/01/2028 27/07/2028 90733
15th year 28/10/2024 50989 4320 28/10/2024 27/01/2029 27/01/2030 27/01/2029 27/07/2029 90734
16th year 28/10/2024 50989 7200 28/10/2024 27/01/2030 27/01/2031 27/01/2030 27/07/2030 90735
17th year 28/10/2024 50989 7200 28/10/2024 27/01/2031 27/01/2032 27/01/2031 27/07/2031 90736
18th year 28/10/2024 50989 7200 28/10/2024 27/01/2032 27/01/2033 27/01/2032 27/07/2032 90737
19th year 28/10/2024 50989 7200 28/10/2024 27/01/2033 27/01/2034 27/01/2033 27/07/2033 90738
20th year 28/10/2024 50989 7200 28/10/2024 27/01/2034 27/01/2035 27/01/2034 27/07/2034 90739

Documents

Orders
Application Number Applicant Section Controller Decision Date
267/MUM/2015
Application Documents
Name Date
1 267-MUM-2015-RELEVANT DOCUMENTS [24-09-2024(online)].pdf 2024-09-24
2 267-MUM-2015-PROOF OF ALTERATION [23-09-2024(online)].pdf 2024-09-23
3 267-MUM-2015-RESTORATION PAYMENT LETTER-20-09-2024.pdf 2024-09-20
4 267-MUM-2015-FORM-15 [02-11-2023(online)].pdf 2023-11-02
5 267-MUM-2015-POWER OF AUTHORITY [02-11-2023(online)].pdf 2023-11-02
6 267-MUM-2015-RELEVANT DOCUMENTS [02-11-2023(online)].pdf 2023-11-02
7 267-MUM-2015-IntimationOfGrant06-05-2022.pdf 2022-05-06
8 267-MUM-2015-PatentCertificate06-05-2022.pdf 2022-05-06
9 267-MUM-2015-Response to office action [04-05-2022(online)].pdf 2022-05-04
10 267-MUM-2015-Written submissions and relevant documents [14-01-2022(online)].pdf 2022-01-14
11 267-MUM-2015-Annexure [14-12-2021(online)].pdf 2021-12-14
12 267-MUM-2015-Correspondence to notify the Controller [14-12-2021(online)].pdf 2021-12-14
13 267-MUM-2015-US(14)-ExtendedHearingNotice-(HearingDate-30-12-2021).pdf 2021-12-07
14 267-MUM-2015-Written submissions and relevant documents [14-08-2020(online)].pdf 2020-08-14
15 267-MUM-2015-Annexure [30-07-2020(online)].pdf 2020-07-30
16 267-MUM-2015-Correspondence to notify the Controller [30-07-2020(online)].pdf 2020-07-30
17 267-MUM-2015-US(14)-HearingNotice-(HearingDate-31-07-2020).pdf 2020-06-30
18 267-MUM-2015-ABSTRACT [10-06-2019(online)].pdf 2019-06-10
19 267-MUM-2015-CLAIMS [10-06-2019(online)].pdf 2019-06-10
20 267-MUM-2015-COMPLETE SPECIFICATION [10-06-2019(online)].pdf 2019-06-10
21 267-MUM-2015-CORRESPONDENCE [10-06-2019(online)].pdf 2019-06-10
22 267-MUM-2015-DRAWING [10-06-2019(online)].pdf 2019-06-10
23 267-MUM-2015-FER_SER_REPLY [10-06-2019(online)].pdf 2019-06-10
24 267-MUM-2015-OTHERS [10-06-2019(online)].pdf 2019-06-10
25 267-MUM-2015-FER.pdf 2019-01-31
26 267-MUM-2015-Form 1-270115.pdf 2018-08-11
27 267-MUM-2015-Form 18-270115.pdf 2018-08-11
28 267-MUM-2015-Form 2(Title Page)-270115.pdf 2018-08-11
29 267-MUM-2015-Form 26-270115.pdf 2018-08-11
30 267-MUM-2015-Form 3-270115.pdf 2018-08-11
31 267-MUM-2015-Form 5-270115.pdf 2018-08-11
32 267-MUM-2015-Form 9-270115.pdf 2018-08-11
33 267-MUM-2015-Other Patent Document-270115.pdf 2018-08-11
34 ABSTRACT1.jpg 2018-08-11
Search Strategy
1 267MUM2015_30-01-2019.pdf

Specification

FORM 2
THE PATENT ACT 1970
(39 of 1970)
&
The Patents Rules, 2003
COMPLETE SPECIFICATION
(See Section 10, and rule 13)
TITLE OF INVENTION ‘
ESSENTIAL OIL COMPOSITIONS FROM PANCHA TULASI DROPS
APPLICANT(S)
a) Name-
b) Nationality
c) Address
AND
a) Name
b) Nationality
c) Address
The following specification particularly describes the invention
BABUULAL BHAVARLAL JAIN
INDIAN National
555 GREEN PARK APARTMENT,
DR. PRADHAN MARG, CHARAI,
THANE (WEST), 400601,
MAHARASHTRA, INDIA
SURESHKUMAR BABUULAL JAIN
INDIAN National ‘
555 GREEN PARK APARTMENT,
DR. PRADHAN MARG, CHARAI,
THANE (WEST)-400601
MAHARASHTRA, INDIA
PREAMBLE TO THE DESCRIPTION
and the manner in which it is to be performed : —
E
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'55:‘.-.
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27-Jan-2015I1718I267-MUM-2015IForm
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FIELD OF THE INVENTION
This invention relates to a simple and natural method for disinfection of potable drinking -
water, by using formulation of an essential oil derived from a herbal material having
predominant antimicrobial and anti-oxidant property. More particularly, this invention
-relates to a five species of basils emulsion preparation, which finds application in
disinfection of drinking water contaminated with harmful pathogens to make it potable at
the same time it provides free radicle scavenging and natural immunity enhancing effects.
BACKGROUND OF THE INVENTION
The health burden of poor water quality is enormous. It is estimated that around 37.7
million Indians are affected by waterborne diseases annually, 1.5 million children are
estimated to die of diarrhoea alone and 73 million working days are lost due to
waterborne disease each year. The resulting economic burden is estimated at $600 million
a year. Poor water quality spreads disease, causes death and hampers socio-economic
progress. Around five million people die due to waterborne diseases. In addition, these
diseases affect education and result in loss of work days, estimated at 180 million person
_ days annually.
The type of treatment and disinfection employed can vary according to the desired
application and regional standards for water reuse. Treatment options for grey water
reuse include coarse filtration (March et al., 2004), direct membrane filtration (Ramon et
' al., 2004), constructed wetland (Dallas and Ho, 2005), rotating biological contactor (Nolde,
1999), and membrane bioreactor (Jefferson et al., 2000). Various chemical are used
including chlorine a leading contender for the disinfection of drinking water however,
there are drawbacks of various synthetic or chlorine applications; which include the
formation of disinfection by-products that can cause adverse health effects (Morris et al.,
1992;
Nieuwenhuijsen et al., 2000), and limited efficacy in the presence of particles and organics
(Winward et al., 2007). Alternative disinfectants that are perceived as being
‘environmentally friendly’ would be an attractive option for end point disinfection of
potable drinking water. Such alternatives include ultraviolet (UV) light and plant
essential oils.
There is thus a need for a simple,‘ inexpensive and widely available purification and
. disinfecting system for potable liquids, especially water, that is: effective against range of
. Gram positive and Gram negative bacteria, viruses and other "microorganisms; safe and
free from the side-effects caused by chemical disinfectants; cost-effective and affordable;
simple to use and can be carried anywhere; and requires no special assistance, equipment
or training to use.
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27-Jan,-2015I1718I267-MUM-2015IForm
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OBJECTS OF THE INVENTION
It is the principal object of the present invention to provide a method and
composition from five species of Tulasi (basils); Pancha Tulsi Drops for disinfecting water
contaminated with pathogenic microorganisms.
It is another object of the present invention to provide pure drinking water, by purifying
water at a very affordable cost.
‘ It is another object of the present invention to provide anti-oxidant and
immunomodulation effect.
SUMMARY OF THE INVENTION
Accordingly, the present invention provides a method of disinfecting water contaminated
with pathogenic microorganisms, comprising treating the contaminated potable drinking
water instantly with an effective amount of a composition enabling the water for end
point use comprising:
a) an emulsifier;
b) an essential oil composition selected from the group consisting of basil oils Ocimum
sanctum; Ocimum basi1icum;_ Ocimum grattissimum, Ocimum canum, Ocimum
citriodorum, Ocimum tenuiflorum L, Ocimum ciliatum, Ocimum kilimandscharicum,
Ocimum americanum; Ocimum campechianum, Ocimum Viride
c) optionally a carrier oil comprising a" plant oil;
optionally a natural anti-oxidant present in basil preserving the shelf life as well as
provide anti-oxidant effect; -
e) optionally containing-an ingredient(s) enhances natural immunity.
' The present invention also provides a composition for disinfecting water contaminated
with pathogenic microorganisms comprising:
a) an emulsifier;
b) an essential oil composition selected from the group consistingof basil oils of Ocimum
sanctum; Ocimum basilicum; Ocimum grattissimum, Ocimum canum, Ocimum
citriodorum, Ocimum tenuiflorum L, Ocimum ciliatum, Ocimum kilimandscharicum,
Ocimum americanum; Ocimum campechianum, Ocimum viride
c) optionally a carrier oil comprising a plant oil;
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27-Jan-2015I1718I267-MUM-2015IForm
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d) optionally a natural anti-oxidant present in basil preserving the shelf life as well as
provide anti-oxidant effect; A ~ '
e) optionally containing an ingredient(s) enhances natural immunity.
I BRIEF DESCRIPTION OF THE ACCOMPANYING DRAWINGS
Figure 1 is a graphical representation of Lipoxygenase inhibition of DMSO, Pancha
Tulasi Drops and Indomethacin.
Figure 2 is a graphical representation of immunomodulation study and assay of lung
homogenate at different concentrations Pancha Tulasi Drops.
DETAILED DESCRIPTION OF THE INVENTION
Certain preferred - embodiments are herein described with reference to the
accompanying drawings. It is to be understood that all these are non-lirniting and the
invention embraces legitimate developments within the ambit of the present invention.
‘Essential oils are aromatic liquids extracted from plant material, typically by steam
distillation. _Essential oils and their components have been shown to possess
antibacterial, antiviral,’ antiparasitic, antifungal, insecticidal and herbicidal properties.
Research into the antimicrobial properties of Essential oils and their components has been
- largely directed towards theiruse as food preservatives.
Essential oils and their components have been shown to inhibit bacterial growth inlmeat,
-fish, and vegetable products. The principal mechanism of "bacterial inactivation by
Essential oils is considered to be the disruption of the cell membrane, causing
leakage of cell contents and eventual cell lysis . This action of certain Essential oils can be
explained by the presence of phenolic compounds, common in many EOs, such as
carvacrol, thymol, and eugenol, which are known to cause such disturbance of the cell
membrane .The effective use of Essential oils and their components as inhibitors of
microbial growth in foods" points to their potential as residual regrowth inhibitors in
_ potable drinking water.
Essential oils are extensively used medicinally in traditional medicine across the world ..
for their great variety of bio-dynamic actions. Besides antimicrobial and antiseptic
_ properties, they are "reported to be antiviral and antitoxic. Essential oils have long been
used as medicines, traditional texts suggest its use as an appetizer,’ digestive cordial,
antitissusive and as mouth freshener. Indications are toothache (local application),
indigestion/ flatulence, nausea, halitosis and hyperacidity. It is also used as anesthetic
and antiseptic in dentistry.
Essential oils are used as vital constituents of many cosmetic preparations, as spices, and
in medicines. In cosmetics essential oils are mainly responsible for fragrance; in spices,

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essential oils are the principle ingredients which contribute to the flavor, easy digestibility
and (sometimes) to the durability of food.
Chemically and biogenetically, the natural product class ”Essential oils” is heterogeneous.
Mono-, sesqui- and di-terpenoids (mevalonic acid-derived constituents);
phenylpropanoids (cinnamic acid-derived compounds) and alkane derivatives (alkanes,
alkenes, alkynes, alkanols, alkanals, alkanoic acids: mostly acetogenins) are by far the
most ubiquitous components of essential oils. Essential oils belong to the so-called
”Natural Products” or secondary metabolites of plants.
. Use of Essential Oils in replacing the chemical/ synthetic disinfectant may reduce the
synthetic residue related adverse effects on body and may help to reduce residue burden
on the environment.
Pancha Tulasi Drops are, a novel composition of Essential Oils derived from 5 species of
basil from the species of; namely Ocimum sanctum; Ocimum basilicum; Ocimum
grattissimum, Ocimum canum, Ocimum citriodorum, Ocimum tenuiflorum L, Ocimum
ciliatum, Ocimum kilimandscharicum, Ocimum americanum; Ocimum campechianum,
Ocimum Viride and blending them to make an emulsion in a specific way to provide
disinfection effect on potable drinking water and providing relief from oxidative stress
(anti-oxidant effect). The blends were made as follows:
# Composition 1 w/w%
1 Ocimum sanctum . . 600
2 Ocimum basilicum 5
3 Ocimum grattissimum 5
4 Ocimum canum 5
5 Ocimum citriodorum ' 5
6 Emulsifier, Solublizer, natural preservatives & Antioxidant 20
# Composition 2 , w/ w%
1 Ocimum sanctum . . _ , ' . . 20
2 Ocimum basilicum ' 5
3 Ocimum grattissimum 5
4 Ocimum canum _ 5
5 Ocimum citriodorum _ 5
6 Emulsifier, Solublizer, natural preservatives & Antioxidant . 70
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Composition 3 w/ w%
1 Ocimum sanctum 10
2 Ocimum basilicum 10
3 Ocimum grattissimum 5
4 Ocimum canum 5
5 Ocimum citriodorum _ 5
6 Emulsifier, Solublizer, natural preservatives & Antioxidant 65
Composition 4 w/ w%
1 Ocimum sanctum ' 10
2 Ocimum basilicum 10
3 Ocimum grattissimum 5
4 Ocimum canum 3
5 Ocimum citriodorum 2
6 Emulsifier, Solublizer, natural preservatives & Antioxidant 70
Composition 5 w/ w%
1 Ocimum sanctum 5
2 Ocimum basilicum 5
3 Ocimum grattissimum 5
4 Ocimum canum 3
5 Ocimum citriodorum 2
6 Emulsifier, Solublizer, natural preservatives & Antioxidant 80
Composition 6 w/ w%
1 Ocimum sanctum 10
2 Ocimum basilicum A 10
3 Ocimum grattissimum 10
4 Ocimum canum .10
5 Ocimum kilimandscharicum 20
6 Emulsifier, Solublizer, natural preservatives &: Antioxidant 40

27-Jan-2015I1718I267-MUM-2015IForm
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# . Composition 7 w/w%
1 Ocimum sanctum 20
2 Ocimum basilicum 5
3 Ocimum grattissimum 5
4 Ocimum canum 5
5 Ocimum kilimandscharicum 5
6 Emulsifier, Solublizer, natural preservatives & Antioxidant 70
# Composition 8 w/ w%
1 Ocimum sanctum 10
2 Ocimum basilicum 10
3 Ocimum grattissimum 5
4 Ocimum canum 5
5 Ocimum kilimandscharicum . V 5
6 Emulsifier, Solublizer, natural preservatives & Antioxidant 65
# Composition 9 w/w%
1 Ocimum sanctum 10
2 Ocimum basilicum 10
3 Ocimum grattissimum 5‘
4 Ocimum canum 3
5 Ocimum kilimandscharicum -_ 2
6 Emulsifier, Solublizer, natural preservatives 8: Antioxidant 70
# Composition 10 W/w%
1 Ocimum sanctum 20
2 Ocimum basilicum 10
3 Ocimum grattissimum . 5
4 Ocimum canum 5
5 _ Ocimum americanum R 3 5
» 6 Emulsifier, Solublizer, natural preservatives & Antioxidant 55

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# Composition 11 w/w%
1 Ocimum sanctum 60
2 Ocimum basilicum 10
3 Ocimum grattissimum 5
4 Ocimum canum 3
5 Ocimum americanum 2
6 Emulsifier, Solublizer, natural preservatives & Antioxidant 20
# Comgasition 12 w/w%
1 Ocimum sanctum 5
2 Ocimum basilicum ' 5
3 Ocimum grattissimum 5
4 Ocimum canum 3
5 Ocimum americanum , 2
6 Emulsifier, Solublizer, natural preservatives & Antioxidant 80
# Composition 13 w/ w%
1 Ocimum sanctum 5
2 Ocimum basilicum 5
3 Ocimum grattissimum 5
‘ 4 Ocimum canum 3
5 Ocimum campechianum ' 2
6 Emulsifier, Solublizer, natural preservatives & Antioxidant 80
# Composition 14 w/ w%
1 Ocimum sanctum 5
2 Ocimum basilicum 5 «
3 Ocimum grattissimum 5
4 Ocimum canum 3
5 Ocimum Viride ' , 2
6 Emulsifier, Solublizer, natural preservatives 8: Antioxidant 80

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# Composition 15 w/ w%
1 Ocimum sanctum 5
2 Ocimum basilicum 5
3 Ocimum grattissimum 5
4 Ocimum ciliatum 3
5 Ocimum kilimandscharicum 2
6 Emulsifier, Solublizer, natural preservatives 8: Antioxidant 80
# Composition 16 w/w%
1 Ocimum sanctum 5
2 Ocimum basilicum 5
3 Ocimum grattissimum 5
4 Ocimum americanum 3
5 Ocimum campechianum _ 2
6 Emulsifier, Solublizer, natural preservatives & Antioxidant 80
# Composition 17 w/w%
1 Ocimum sanctum 10
a 2 Ocimum basilicum 5
3 Ocimum grattissimum 5
4 Ocimumcagchianum 3
5 Ocimum Viride 2
6 Emulsifier, Solublizer, natural preservatives & Antioxidant 75
# Composition 18 w/w%
1 Ocimum sanctum 5
2 Ocimum basilicum 5
3 Ocimum grattissimum 5 '
4 Ocimum kilimandscharicum 3
5 Ocimum americanum 2
6 Emulsifier, Solublizer, natural preservatives &: Antioxidant 80

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# Composition 19 w/w%
1 Ocimum sanctum 5
2 Ocimum basilicum 5
3 Ocimum grattissimum 5
4 Ocimum americanum 3
5 Ocimum campechianum 2
_ 6 Emulsifier, Solublizer, natural preservatives & Antioxidant 80
# Composition 20 w/ w%
1 Ocimum sanctum R 5
2 Ocimum basilicum 5
3 Ocimum grattissimum 5
4 Ocimum campechianum 3
5 Ocimum Viride . 2
6 Emulsifier, Solublizer, natural preservatives 8: Antioxidant 80
The genus Ocimum involves economically the most important medicinal and aromatic
herbs, under shrubs or shrubs in the world. It belongs to the family Lamiaceae, subfamily
Ocimoideae, and comprises more than_30 species distributed in tropical and subtropical
. regions of Asia, Africa, and Central and South America (Paton 1992). Traditionally, the
genus Ocimum is widely used for the treatment of various ailments including
rheumatism, paralysis, epilepsy, high fever, diarrhea, sunstroke, influenza, gonorrhea,
mental illness, abdominal pains, colds, coughs, measles, and has also antipyretic,
, antihelmentic, stomatic, anti-emetic, and antimalarial effects (Caceres et al. 1990; Obeng-
Ofori et al. 1998; Nyarko et al. 2002; Ezekwesili et al. 2004). It is also a source, of aroma
compounds and essential oils containing biologically active constituents that possess
insecticidal (Deshpande et al. 1997), nematicidal (Chaterjee et al. 1982), and fungistatic
properties (Reuveni et al. 1984). The active compounds present as volatile oil from the
leaves consist mainly of eugenol, thymol, citrol, geraniol, camphor, linalool l, and methyl
cinnamate (Charles &: Simon 1992; ]irovetz&:Buchbauer 2001; Mondello et al. 2002; Vina&:
Murillo 2003; Padalia& Verna 2011; Singh et al. 2011 ; Verma et al. 2011). v
The seeds contain oil composed of fatty acids and sitosterol. The roots contain sitosterol
and three triterpenes A, B, and C. Additionally, they also contain rosmarinic acid, thymol,
‘ methyl chavicol and, citral etc. (Dhar et al. 1968), and vitamins C, A, and minerals like
calcium, zinc, and iron (Anbarasu&Vijayalakshmi 2007), as well as chlorophyll andmany
other phytonutrients.However, the antimicrobial activity of Ocimum essential oil against
microorganisms has been investigated by some researchers (Prasad et al. 1986; N akamura
- et al. 1999; Adebolu&Oladimeji 2005; Adigiizel et al. 2005; Moghaddam et al. 2011 ; Verma
et al. 2011) using different techniques and their investigations mostly covered one
individual or two species.
n,_
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ii 2:‘ I
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A The antimicrobial activities of PanchaTulasi Drops may be due to high contents of tannins
and phenolic constituents. The most active constituents (essential oils) rich in phenolic
compounds are widely reported to possess high levels of antimicrobial activity (Prasad et
al. 1986; Nakamura et al. 1999; Dorman & Deans 2000).
Essential oils and their components have been shown to possess antibacterial, antiviral,
antiparasitic, antifungal, insecticidal, and herbicidal properties. Research into the
antimicrobial properties of Essential oils and their components has been largely directed
towards their use as food preservatives. Essential oils and their components have been
shown to inhibit bacterial growth in meat, fish, and vegetable products. The principal
mechanism of bacterial inactivation by Essential oils is considered to be the disruption of
the cell membrane, causing leakage of cell contents and eventual cell lysis. This action of
certain Essential oils can be explained by the presence of phenolic compounds, common
in many EOs, such as carvacrol, thymol, and eugenol, which are known to cause such
disturbance of the cell membrane.
The genus Ocimum involves economically the most important medicinal and aromatic
herbs, under shrubs or shrubs in the world. It belongs to the family Lamiaceae, subfamily
Ocimoideae, and comprises more than 30 species distributed in tropical and subtropical
regions of Asia, Africa, and Central and South America. Traditionally, the genus Ocimum
is widely used for the treatment of various ailments including rheumatism, paralysis,
epilepsy, high fever, diarrhea, sunstroke, influenza, gonorrhea, mental illness, abdominal
pains, colds, coughs, measles, and has also antipyretic, antihelmentic, stomatic, anti-
emetic, and antimalarial effects. It is also a source of aroma compounds and essential oils
containing biologically active constituents that possess insecticidal, nematicidal, and
.fungistatic properties. The active compounds present as volatile oil from the leaves
consist mainly of eugenol, ‘thymol, citrol, geraniol, camphor, linalool l, and methyl
cinnamate. I
_ The seeds contain oil composed of fatty acids and sitosterol. The roots contain sitosterol
and three triterpenes A, B, and C. Additionally, they also contain rosmarinic acid, thymol,
methyl chavicol and, citral etc., and vitamins C, A, and minerals" like calcium, zinc, and
iron as well as chlorophyll and many other phytonutrients. However, the antimicrobial
activity of Ocimumlessential oil against microorganisms has been investigated by some
researchers.
The effective use of Essential oils and their components as inhibitors ofmicrobial growth
in foods points to their potential as residual regrowth inhibitors in potable drinking
water. PanchaTulasi Drops are, a novel composition of Essential Oils derived from 5
species of basil from the following species of Ocimum sanctum; Ocimum basilicum;
' Ocimum grattissimum, Ocimum canum, Ocimum citriodorum, Ocimum tenuiflorum L,
Ocimum ciliatum, Ocimum kilimandscharicum, Ocimum americanum; Ocimum
campechianuma and Ocimum virideand blending them to make an emulsion in a specific
Way to provide disinfection effect on potable drinking water and providing relief from
» oxidative stress (anti-oxidant effect).

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M ‘"121 cT...TI£
. medium)
‘-r 5 Y . ..
3"‘: 5-.3‘ ‘FE 3
The antimicrobial activities of Pancha Tulasi Drops may be due to high contents of
tannins and phenolic constituents. The most active constituents (essential oils) rich in
- phenolic compounds are widely reported to possess high levels of antimicrobial
activity.
In an in-vitro Aromatic oil Challenge test was performed on Pancha Tulasi Drops;
the Culture suspension was standardized to get a cell count between 70-100 cfu/ ml. To
each of the tube of distilled water, variable quantities of drops of aromatic oil were added
to each of the tube of distilled water.
A quantity of enumerated culture suspension of micro- organism was added to 200 ml of
the Distilled water containing Aromatic oil_so that the final count was NMT 100 cfu/ ml.
_ The contact time was kept for 5 minutes. After completion of the Contact time 1 ml of the
Inoculated Distilled water was plated out .Bulk seed with SCDA media containing 0.1%
Soyalecithin and 1% Tween 80. (2 plates for every concentration).The Plates were
incubated at 30- 35 deg Centigrade for 24-48 hours. The plate counts were read after 24-48
. hours respectively. The results were Interpreted For Every concentration.
The results showed that Panchatulasi Oil Liquid Drops is effective against
Staphyllococcus and Salmonella Typhi infection.
Table 1: Anti -Microbial activity of PanchaTulasi liquid drops on Salmonella Typhi I
~
~ ~
~
~ ~
Pr
no Quantity .1 3.1 .2 1.2 3.2 1.3 K, R,
1 1 drop Nil , Nil Nil il Nil Nil
2 5 drop Nil il Nil Nil Nil -Nil
3 10 drop Nil Nil . Nil Nil Nil Nil
Table 2: Anti -Microbial activity of Panchatulasi Liquid Drops on Staphylococcus
Aureus
Sr 'Samplel C C ‘Al C5 l l
, W 3 l .
11 drop, Nil Nil Nil Nil Nil Nil Nil Nil
25 drop Nil Nil Nil V Nil Nil Nil Nil Nil
310 drop il Nil Nil Nil Nil Nil Nil ' Nil
BC= Batch
In vitro Minimum Inhibitory Concentration (MIC) of the Pancha Tulasi Drops was
- performed using micro broth dilution method against Gram positive and Gram negative
bacterial ATCC 49619 Streptococcus pneumoniae strains_ and ATCC 700904
Streptococcus pneumonia respectively. Panch Tulasi "Oil variants in different
dilution were weighed and dissolved using 50% DMSO solution (DMSO + MIC broth
and subsequent dilutions were made with broth. Pancha Tulasi Drops were
1 _—.- + . .- —. . . . . . =~ —~, .r _=—
s..:; L; , ._ , . ._ . _ , . .1. .- 2.. 2 .= :3 .= :_,_
'...-' =‘ '. J. .; 1 =»_~‘ _L ..:_ L.) _._ ._" J; _L __.‘-
12

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"71 .:":_=‘:x
dissolved using 60% DMSO solution, and further dilutions are made with broth. The
reference compounds Ciprofloxacin, Vancomycin were dissolved in water as per the CLSI
guidelines. The compound dilution was done as per CLSI guidelines. One hundred and
seventy five microliters of CAMHB was added to the first column of a 96 well flat bottom
plate, which is the media control. Second column is for the stocks of the test compounds
and reference standards from which dilutions are made. Then 175 uL of medium was
added to the 5th, 8th and 11th columns, 75 uL of medium to the 4th, 7th and 10”‘ columns
and 50 uL of medium to the 3rd, 6”‘ and 9th columns and the 12”‘ column was the organism
control. Four milligram per mL stocks of the test Pancha Tulasi Drops and further
dilution was made from this stock. Eight mg/mL stocks of the Pancha Tulasi Drops were
made and further dilution was made from‘ this stock Two mg/mL stocks of the reference
compounds (Ciprofloxacin and Vancomycin) and further dilution was made from this
stock. The bacterial colonysuspension was made (0.9 to 1.0 McFarland’s standard) using
V Densimat. Later it was diluted 100 times in CAMHB.
One hundred microliters of diluted culture was added to a micro titer plate, containing
100 |.1l_. of diluted compound or media, such that the final inoculum was 0.5 to 1 X 106
CFU/mL and the drug concentration was halved. All inoculated plates except
' Streptococcus pneumoniae were incubated at 35 °C for 24 hours.
Streptococcus pneumonia plates were incubated in incubator supplied with 5% CO2‘at 35
°C for 24 hours. The assay plates were read visually for the growth inhibition and also at
600 nm wavelength.
Sr No lCompounds ATCC 49619 ATCC 700904
- I Streptococcus ‘Streptococcus
neumoniae neumoniae
1 Pancha Tulasi 32 ug/ mL 64ug/ mL
variants
2 Pancha Tulasi 128ug/ mL 256ug/ mL
Drops «
3. Ciprofloxacin 1 ug/ mL 2 pg/mL
Vancomycin 0.25 ug/mL , 0-.25 pg/mL
_ 49619 Streptococcus
Table 3: Minimum inhibitory concentration for Pancha Tulasi oil variants,
PanchaTulasi Drops Marketed compound, ciprofloxacin and Vancomycin
The overall resul indicates that Pancha Tulasi Drops variants showed MIC of 32 pg/mL
and 64 lug/mL against ATCC 49619 Streptococcus pneumonia and ATCC 700904
Streptococcus pneumonia.‘ PanchaT ulasi ‘drops showed. MIC of 128ug/mL and 256
ug/ mL.Marketed compound showed MIC of 2048 ug/ mL and
1024ug/mL.ciprofloxacin showed MIC of 1 ug/mL and 2 ug/mL against ATCC
pneumonia and ATCC) 700904 Streptococcus
pneumonia.Vancomycin showed 0.25 pg/mL and 0.25 pg/mL. '

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In furtherance; an Immuno- modulatory effect of Pancha Tulasi Drops were studies in
mouse splenocytes 2 mg/mL of stock solution was prepared in PBS and stored at -80 °C.
The working stock solution (200 ug/mL) was prepared from initial stock in RPMI-1640
medium. The spleen was removed from male BALB/c Mice aged 6 -7 weeks (22 - 26
' grams). The spleens were placed into sterile culture dish containing plain RPMI-1640
medium. The splenocytes was extracted using cell strainer (70 um) and centrifuged at 320
X g for 4 minutes at 4 °C. The supernatant was discarded and pellet re-suspended with 1
mL of RBC lysis buffer (ACK lysis) and incubated at RT for 10 minutes. After incubated
1 ACK buffer was removedby washing twice with plain RPMI and cells counted in
hemocytometer. Finally, pellets were re-suspended in RPMI medium supplemented with
2 mM glutamine, 10% heat-inactivated FBS and NaHCO3. The counted cells were diluted
with fresh RPMI medium to get 2 X 106 cells/mL and 180 uL of cells suspension was
added to the wells. Ten rnicroliter of diluted LPS was added to 180 uL of cell suspension
to test wells. The RPMI-1640 medium was added to control wells (No induction). Test
compound (10 uL) from final stock solution (test compound B. No: 2.3) was added to 180
pL of cell suspension and LPS mix, incubation wells to obtain 100, 10, 1, 0.1, 0.01, 0.001
and 0.001 ug/mL. Thefinal concentration DMSO was 0.25%. The reference compounds
(10 uL) of final stock solution was added into 180 uL of cell suspension and LPS mix, in A
A incubation wells to obtain 10, 1, 0.1, 0.01 and 0.001 uM).The final DMSO concentration in
plate about 0.5%. For control wells, ten micro liter of RPMI medium containing 5% DMSO
(No inhibition). After addition of test compounds, ‘plates were incubated in 5 % CO2
incubator for a period of 72 hours. After 24, 48 and 72 hours of incubation, plates were
centrifuged at 2000 RPM at 4 °C for 10 minutes. The supernatant was removed (140 uL)
0 without disturbing cell pellet and stored at -20 °C for cytokine estimation. The cell pellet
was taken for viability assay.
The cell viability of test compound was assessed by the mitochondrial dependent
reduction of 3-(4, 5-dimethythiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) to
purple formazan. The cell suspension test compound mixture, were incubated with MTT
(5 mg/mL) for 2 hour at 37 °C. The medium was removed by aspiration and formazan
crystals were dissolved in DMSO. The extent of the reduction of MTT was quantitated by
measurement of the absorbance at 570 nm. ‘ ' '
-Table 4: Cytokine inhibition level of Pancha Tulasi Drops, Paclitaxel and
1......
lg.
‘“‘lii‘i
Dexamethasone
Compounds Concentration TNF-a(°/o IL-6 (“/0
Inhibition inhibition)
. hours 72 hours
Pancha Tulsi ' 10 ‘ -42.38 37.52
Drops(ug/mL) 100 10.31 98.92
B 2.3
Paclitaxel(uM) 1 . 9 -18.09 ;50.54
10 4 -40.97 . ‘ -54.2
Dexamethasone 1 47.2 . 90.9
’ 10 42.65 89.66
.. - ._ . .- - 14)

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trwiv
'*:::r_
. Compounds Concentration 72 hr °/o viability
Pancha Tulsi Drops
10 .7
(ug/mL) 12° 5
B 2.3 100 33.96
_ 1 110.69
Pachtaxe|(pM)
10 ‘ 78.62
Dexamethasone(pM) :0
Table 5. Cytotoxicity results of Pancha Tulasi Drops ,Paclitaxel and Dexamethasone
Pancha Tulasi Drops showed 37.52 % inhibition of IL-6 at 72 hours .It did not inhibit
TNF-a at 10 ug/mL concentration. At 100 ug/mL Pancha Tulasi drops showed
10.31% & 98.92% for TNF — CI and IL- 6 concentration respectively. Paclitaxel showed no
inhibition in % of TNF '- aand IL-6 respectively. These results indicates superior
immonomodulating activity of Pancha Tulasi Drops with least effect on cell viability in a
higher dosage forms.
‘The anti-inflammatory activity of Pancha Tulasi Drops was found in an in-vitro
Lipoxygenase inhibitory study. Lipoxygenase inhibitor screening kit (Cat No:
760700)was used in the study. Compound stock (10 mg/mL) solutions were prepared
in 100% DMSO and 1:1 methanol water. Pancha Tulasi Drops solution dissolved in 1:1
I DMSO to get 50% DMSO final concentration buffer (100111) is added to wells assigned as
blank wells. 100 um of Indomethacin and Asprin(10p.L) is added to wells assigned as
positive control. For 100% initial activity wells, Lipoxygenase enzyme(90 uL) and solvent
(10p.L) were added .For Inhibitor wells Lipoxygenase enzyme and inhibitor (10uL
sample) were added. The reaction is initiated by adding substrate (10pL arachidonic
acid) and the plate is incubated on a shaker for 5 minutes. After incubation,
chromogen(100uL) was added to each well to stop enzyme catalysis and develop the
reaction. The plate was covered and incubated for five minutes on a shaker. After
incubation, the absorbance was at read 500nm using a plate reader.
‘Table 6: Lipoxygenase °/o inhibition for DMSO, Pancha Tulasi and Indomethacin
Test compound % Inhibition
DMSO . 30
Pancha Tulasi 10 58
mg/mL
Indomethacin 100pM 86
The accompanying figure 1 graphically represents Lipoxygenase inhibition of
DMSO, Pancha Tulasi and Indomethacin.
Pancha Tulasi Drops showed 58% inhibition compared to 30% inhibition shown by
DMSO. In Vitro Antibacterial efficacy studies were conducted for Pancha Tulasi Drops for
varying doses viz 150 mg/bw, 75mg/ KG Bw , 37.5 Kg bw, 18.74 mg/ Kg bw, 9.36 mjg/ kg
.~.~s» .-—- =- —.
»._,r':
~~~
- ‘ C3 .- -. _i_ ..-’ i .5 4-

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I
;t,...;. "'21":
bw, 9.36 mg/ kg bw and reference compound 10 mg/ kg bw on neutropenic mouse model.
Mice were housed in groups of 6 animals per cage, and allowed access to feed and
water ad libitum under controlled conditions. Mice were acclimatized for a minimum
A of six to seven days prior to the study. The animals were observed daily for general health
during this period. Animals were rendered neutropenic. The bacterial strain ATCC
700904 Streptococcus pneumoniae was cultured in Sheep blood agar and incubated at 5%
CO2 incubator for overnight at 35 °C. The culture was aseptically transferred to a tube of
normal saline and the optical density was adjusted using Densimat instrument. The
' culture was diluted to provide a challenge inoculum, and volume of 20 uL was injected at
time "0" through intranasal route. The challenge inoculum count was estimated prior to
and after infection by modified Miles and Misra method, and the animals were treated as
per the study protocol. The test compound B No. 2.3 was administered three times daily
for a period of 3 days by oral route. The reference compound Amoxicillin was
administered once daily for a period of 3 days by oral route. The untreated group was
administered with 0.5 % CMC solution by oral route for a period of 3 days. The lung
samples were removed from untreated groups at ”0” hour and 24 hour post treatment.
Lung tissues were removed from the test compound and reference standard treatment
groups after 24 hour post treatment.
The lung tissue samples were immediately homogenized in 1.0 mL of ice-cold
sterilesaline with a Polytron tissue homogenizer (Kinematica, Lucerne, Switzerland).
The viable counts were determined on 5% Sheep blood agar by plating duplicate 20 uL
. samples of serial ten-fold dilutions of the homogenate. As per the literature any
antibacterial synthetic compound showing more than 2 log1o reduction (Ken Bartizalet al,
Activity of Tedizolid Phosphate (TR-701) in Murine Models of Infection with Penicillin-
resistant and Penicillin-sensitive Streptococcus pneumoniae) in organ burden model(s) was
considered as significant Effective Dose 50 (ED5o)16
~
groups .. .. . , . ._ I . . - .....%Log1o
Untreated 0 hour ' 5.22 i 0.25
Untreated 24 hour post treatment 7.55 i 0.45
' B. No:2.3 (15o)* 6.45 1- 0.55
B. No:2.3 (75) 6.87 :1: 0.46
0 B. No:2.3 (37.5) 7.26 i 0.52
B. No:2.3 (18.74) I 7.57 1- 0.41
B. No:2.3 (9.36) 7.58 i 0.41
Ref compound Amoxicillin (10) 5.32 0.62
Table 7
_ --1*_--1:

'
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;.«-..A,. mg:
3. Iiffffil.
The table 7 above shows the Log10 value of The Pancha Tulasi Drops concentrations and
immunomodulation study and assay of lung homogenate "at different concentrations
Pancha Tulasi Drops.
Test compound B. No: 2.3 showed 1.11 log1o reductions at 150 mg/ kg body weight
compared to untreated infection control with a statistical significance of P < 0.05. At
concentration of 75 and 37.5 compound B No. 2.3 exhibited log1o reduction of 0.68 and
0.29 respectively Test compound B No 2.3, did not exhibit log1o reduction at 18.74 and
9.36 mg/ kg body weight concentration. The reference compound Amoxicillin exhibited
A 2.23 log1o reduction_ at 10 mg/ kg body weight when compare with untreated infection
control.
Batch number: 2.3 exhibited significant antibacterial efficacy at 150 mg/ Kg bodyweight
against acute Mouse lung infection.
The five basils from following species of basils Ocimum sanctum; Ocimum basilicum;
Ocimum grattissimum, Ocimum canum, Ocimum citriodorum, Ocimum tenuiflorum L,
Ocimum ciliatum, Ocimum kilimandscharicum, Ocimum americanum; Ocimum
campechianum, Ocimum viride, are standardized for essential oil extraction for
, preparation of Pancha Tulasi Drops. The five species basil essential oils offer promise as
biologically active constituents, in that they confer antibacterial, antifungal and
antioxidant properties. Therefore use of Pancha Tulasi Drops for disinfection of drinking
water can be an effective natural way to reduce the waterborne diseases occurring at end
_ point use. This further reduces the possibility of microbial growth during storage in pot &
unhygienic utensils used for drinking during end point use. At the same time it will
provide a superior anti-oxidant effect.
The method of disinfecting water contaminated with pathogenic microorganisms,
comprises treating the) contaminated potable drinking water instantly with an effective
amount of a composition enabling the water for end point use comprising:
a) an emulsifier;
b) an essential oil composition selected from the group consisting of basil oils Ocimum
sanctum; Ocimum basilicum; Ocimum grattissimum, Ocimum canum, Ocimum
citriodorum, Ocimum tenuiflorum L, Ocimum ciliatum, Ocimum kilimandscharicum,
Ocimum americanum; Ocimum campechianum, Ocimum Viride
Optionally a carrier oil comprising ‘a plant oil.
optionally a natural anti-oxidant present in basil preserving the shelf life as well as
provide anti-oxidant effect '
optionally containing an ingredient(s) enhances natural immunity
The drinking water is added with the composition in the dosage of 1 drop per 200 ml
to drops per 200 ' I V
e»~.r..
Li 1 _ .
e>
‘-_» _=_ -5'; ‘-_-' _E. :.- 3, _..
17

.
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(,,...v1. “:11:
, ester, protocatechuic
Essential oil were extracted as follows, the fresh samples where thoroughly washed, cut
to pieces and used for steam, distillation, the essential oils of Ocimum sanctum; Ocimum
basilicum; Ocimum grattissimum, Ocimum canum, Ocimum citriodorum, Ocimum
tenuiflorum L, Ocimum ciliatum, Ocimum kilimandscharicum, Ocimum americanum;
Ocimum campechianum, Ocimum viridecollected prior and after flowering were both
obtained by hydro distillation of the freshly cut samples, the distillates thus acquired
were extracted with hexane, and or ethanol and or petroleumether, the oils of of Ocimum
sanctum; Ocimum basilicum; Ocimum grattissimum, Ocimum canum, Ocimum
citriodorum, Ocimum tenuiflorum L, Ocimum ciliatum, Ocimum kilimandscharicum,
Ocimum americanum; Ocimum campechianum, Ocimum virideare extracted and
’ blended in a specific portion.
It is also possible that the essential oils were extracted and the oils contain phenolic and
non-phenolic constituents whereas, the phenolic portion contains, carvacrol and eugenol
and the non-phenolic portion contains, eugenol methyl,caryophyllene and various other
chemical compounds.
It is also possible that the essential "oil were extracted and the oils contain phenolic and
non-phenolic constituents whereas, the phenolic portion contains, carvacrol and
eugenol and the non-phenolic portion contains, eugenol methyl,caryophyllene and
various other chemical compounds.
It is also possible that the essential oil were extracted and the oils contains a group
phenolic compounds, rosmarinic acid, gallic acid, gallic acid methyl ester, gallic acid ethyl
acid, vanillicacid, 4-hydroxybenzoic acid, vanillin, 4-
hydroxybenzaldehyde, carnosic acid, caffeic acid and chlorogenic acid,among these
phenolic compounds rosmarinic acid &:caffeic acid are known as the synergistic anti-
oxidant compound in Pancha Tulasi Drops, Rosmarinic acid (RA), an ester of caffeic acid
and 3,4dihydroxyphenyllactic acid, is a natural antioxidant most commonly occurring in
the species of Lamiaceae and Boraginaceae,it is atannin-like compound and originally
found in Rosmarinus officinalis, RA has received particular attention because of its
well-known biological activities including antibacterial, antiviral, and anti-
inflammatory properties, RA was successfully isolated from Thymus vulgaris, Melissa
officinalis.
‘Alternatively, essential oil were extracted and the oils contain non-phenolic
constituents, the main constituents of Ocimum sanctum are Beta Ocimene (0.18-0.30%),
Beta Ocimene&Ocimene (0.61 - 2.10%), Linalool (0.07 - 0.01%), Methyl Chavicol (0.32 -
1.0%), Eugenol (61.70 -72.00%), Methyl Eugenol (0.314 - 0.034%), BetaCaryophellene
‘ (23.25 - 25.29%). Ocimumgrattissimum: Linalool: (1.53 -2.58%), Methyl Chavicol (0.99 -
1.23%), Eugenol (78.00 - 83.00%), Beta Caryophellene (6.91 - 8.15%).
Ocimumcitriodorum: Linalool (5.13 - 9.69%), Methyl Chavicol (0.79 - 0.88%), Citral-2
(7.45% - 8.80%), Eugenol (14.3 -18.75%), Methyl Eugenol (0.17 - 0.22%), Beta
Caryophellene (38.00- 46.88%). Ocimumcanum: Beta Ocimene, (4.08 - 6.66%), Beta
Ocimene&Ocimene (0.10 -0.22%), Linalool (47.00 -56.71%), Methyl Chavicol(0.06 -0.88%),
. ,.._
_ ;,_,s _g_ _. _'§_ 1‘ 3
18

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i---1.
Mr‘: 2,. Ir’
§-‘-‘E E -. -.-‘*1. Em: 1.4.
E! ‘-...»‘ ll L.-' :‘ ‘-.
Eugenol(0.38 -1.96%), Methyl Eugenol (0.22 - 1.3%, Beta Caryophellene (3.31 -
5.43%). Ocimumbasillicum: Linalool (9.72 - 12.18%), Methyl Chavicol (28.98 - 33.25%),
Citral-1 (0.15 - 0.88%), Citral-2 (0.46 - 0.64%), Eugenol (0.72 - 1.36%), Methyl Eugenol (0.03
'- 0.1 %), Beta Caryophellene (33.78 - 46.94%). The contaminating pathogenic
microorganisms are selected from the group consisting of E. coli, Salmonella typhi and
Vibrio cholera, Staphylococcus Aureus by way of example-.
The amount of composition used for disinfecting the pathogen contaminated water is in
the ratio of about 1:10000 to 12200000 of composition to water or this ration may be about
1:10000 to 1:20000 of composition to water or about 1:20000 and 1:200000 of composition
to water.
The essentialoil of 5 speciesof basils inunique variants effectively controlling the bacterial
growth in challenge tests.
The emulsifier may be selected from the group consisting of poloxamers and their
derivatives, polyoxyethylene 50 stearate, polyoxyl 35 castor oil, polyoxyl 10 oleyl ether,
polyoxyl 20 cetostearyl ether, polyoxyl 40 stearate, polysorbate 20, polysorbate 40,
polysorbate 60, polysorbate 80, propylene glycol diacetate, propylene glycol
monostearate, polyethylene glycol, sodium lauryl sulfate, sodium stearate, sorbitan _
mono-laurate, sorbitan mono-oleate, sorbitan mono-palrnitate, sorbitanmonostearate,
stearic acid, ethyl alcohol, Propylene glycol, and emulsifying wax.
‘The solubilizer comprises Polyoxyethylenesorbitan fatty I acid esters (1-15%), 1,2-
Dihydroxypropane (1-10%), a-Hydro-o-hydroxypoly(oxy-1,2-ethanediyl)(1-15%) and its
one or the various polymers.
V It is possible that the emulsifier is present in the composition in the range of about 0.2 to
1.0 % wt. of essential oil.
The solubilizer is present in the composition in the range of about 1 to 15.0 % wt. of
essential oil. '
The anti-oxidants comprises of Rosmarinic acid (RA), an ester ofcaffeic acid, 3,4
dihydroxyphenyllactic acid, tert-butyl-4-methoxyphenol, 2,6-di-tert-butyl-p-cresol, p-
cresol, alpha- bisabolol, carvacrol in the range of about 0.02 - 2 % of the composition.
_ The ratio of contaminated water to emulsion of the essential oil is in the rangebetween
10:1 to 500021.
I
for (disinfecting water contaminated with pathogenic
The composition
microorganisms comprises:
a) an emulsifier;
b) an essential oil composition selected from the group consisting of basil oils Ocimum
sanctum; Ocimumwbasilicum; Ocimgm grattissimum, Ocimum canum, Ocimum
_L.-*"2_.-
19
2 .-' =...~' J. .1 x._.- L

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F-(_-av‘. "Til
~
citriodorum, Ocimum tenuiflorum L, Ocimum ciliatum, Ocimum kilimandscharicum,
Ocimum americanum; Ocimum campechianum, Ocimum Viride;
optionally a carrier oil comprising a plant oil;
optionally a natural anti-oxidant present in basil preserving the shelf life as well as
provide anti-oxidant effect; A
optionally containing an ingredient(s) enhances natural immunity.
~
t,.,.:;
J
;...-«=; I
I I

R
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I'VE CLAIM:
1.
~
A method of disinfecting water contaminated with pathogenic microorganisms,
comprising treating the contaminated potable drinking water instantly with an
effective amount of a composition enabling the water for end point use
comprising:
a) an emulsifier;
b) an essential oil composition selected from the group consisting of basil oils of
Ocimum sanctum; Ocimum basilicum; Ocimum grattissimum, Ocimum canum,
Ocimum citriodorum, Ocimum tenuiflorum L, Ocimum ciliatum, Ocimum
kilimandscharicum, Ocimum americanum; Ocimum campechianum, Ocimum
Viride; '
c) optionally a carrier oil comprising a plant oil;
d) optionally a natural anti-oxidant present in basil preserving the shelf life as well
as_provide anti-oxidant effect; .
e) optionally containing an ingredient(s) enhances natural immunity.
The method of claim 1, wherein the drinking water is added with the composition
in the dosage of 1 drop per 200 ml to 5 drops per 200 ml.
The method of claim 1, wherein essential oil were extracted as follows, the fresh
samples where thoroughly washed, cut to pieces and used for steam, distillation,
the essential oils of Ocimum sanctum; Ocimum basilicum; Ocimum grattissimum,
Ocimum canum, Ocimum citriodorum, Ocimum tenuiflorum L, Ocimum ciliatum,
Ocimum kilimandscharicum, Ocimum americanum; Ocimum campechianum,
Ocimum Viride collected prior and after flowering were both obtained by hydro
distillation of the freshly cut samples, the distillates thus acquired were
extracted with hexane, and or ethanol and or petroleum ether, the oils of of
Ocimum sanctum; Ocimum basilicum; Ocimum grattissimum, Ocimum canum,
Ocimum citriodorum, Ocimum tenuiflorum L, Ocimum ciliatum, Ocimum
kilimandscharicum, Ocimum americanum; Ocimum campechianum, Ocimum
virideare extracted and blended in a specific portion.
The method of claim 1, wherein essential oil were extracted and the oils contain
phenolic and non-phenolic constituents whereas, the phenolic portion contains,
carvacrol and . eugenol and the non-phenolic portion contains, eugenol
methyl,caryophyllene and various other chemical compounds.
The method of claim 1, wherein essential oil were extracted and the oils contain
phenolic and non-phenolic constituents whereas, the phenolic portion contains,
carvacrol and eugenol and the non-phenolic portion contains, eugenol
methyl,caryophyllene and Various other chemical compounds.
The method of claim 1, wherein essential oil were extracted and the oils contains a
group phenolic compounds, rosmarinic acid, carnosic acid, gallic acid, gallic acid
methyl ester, gallic acid ethyl ester, protocatechuic acid, vanillicacid, 4-
hydroxybenzoic acid, vanillin, 4-hydroxybenzaldehyde, caffeic acid and
.. .. 3‘ LI. . . .. .
.:'_i...=rI_.- _-..='j_:.-
21
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6-1.
1“ K
II_.
10.
11.
12.
13.
~~~
chlorogenic acid,among these phenolic compounds rosmarinic acid&caffeic acid
are known as the synergistic anti-oxidant compound in PanchaTulasi Drops,
Rosmarinic acid (RA), an ester of caffeic acid and 3,4dihydroxyphenyllactic acid, is
a natural antioxidant most commonly occurring in the species of Larniaceae and
Boraginaceae, it is atannin-like compound and originally found in
Rosmarinusofficinalis, RA has received particular attention because of its well-
known biological activities including antibacterial, antiviral, and anti-
inflammatory properties, RA was successfully isolated from Thymus vulgaris,
Melissa officinalis.
. The method of claim 1, wherein essential oil were extracted and the oils contain
non-phenolic constituents, the main constituents of Ocimum sanctum are Beta
Ocimene (0.18-0.30%), Beta Ocimene&:Ocimene (0.61 - 2.10%), Linalool (0.07 -
0.01%), Methyl Chavicol (0.32 - 1.0%), Eugenol (61.70 -72.00%), Methyl Eugenol
(0.314 - 0.034%), BetaCaryophellene (23.25 - 25.29%). Ocimumgrattissimum:
Linalool: (1.53 -2.58%), Methyl Chavicol (0.99 - 1.23%), Eugenol (78.00 - 83.00%),
Beta Caryophellene (6.91 - 8.15%). Ocimumcitriodorum: Linalool (5.13 -
9.69%), Methyl Chavicol (0.79 - 0.88%), Citral-2 (7.45% - 8.80%), Eugenol (14.3 -
18.75%), Methyl Eugenol (0.17 -0.22%), Beta Caryophellene (38.00- 46.88%).
Ocimumcanum: Beta Ocimene, (4.08 - 6.66%), Beta Ocimene&Ocimene (0.10 -
0.22%), Linalool (47.00 -56.71%),. Methyl Chavicol(0.06 -0.88%), Eugenol(0.38 -
1.96%), Methyl Eugenol (0.22 - 1.37), Beta Caryophellene (3.31 - 5.43%).
Ocimumbasillicum: Linalool (9.72 - 12.18%), Methyl Chavicol (28.98 - 33.25%),
Citral-1 (0.15 - 0.88%), Citral-2 (0.46 - 0.64%), Eugenol (0.72 - 1.36%), Methyl
Eugenol (0.03 - 0.1 %), Beta Caryophellene (33.78 - 46.94%).
The method as claimed in claim 1, wherein ' the contaminating pathogenic
microorganisms are selected from the group consisting of E. coli, Salmonella
typhi and Vibrio cholera, Staphylococcus Aureus A
The method as claimed in claim 1, wherein the amount of composition used for
disinfecting the pathogen contaminated water is in the ratio of about 1:10000 to
12200000 of composition to water.
The method as claimed in claim 1, wherein the amount’ of composition used for
disinfecting the pathogen contaminated water is in the ratio of about 1:10000 to
1220000 of composition to water.
The method as claimed in claim 1, wherein the amount of composition used for
disinfecting the pathogen contaminated water is in the ratio of about 1:20000 and
1:200000 of composition to water.
The method as claimed in claim 1, wherein the essentialoil of 5 speciesof basils in
unique variants effectively controlling the bacterial growth in challenge tests.
The method as claimed in claim 1, wherein the emulsifier is selected from the
group consisting of poloxamers and their derivatives, polyoxyethylene 50 stearate,
_piol;yoxyla_.__35 _cams___tpr“ oi‘_l,;__:p;_ol;yoxyl playl ether, polyoxyl 20 cetostearyl ether,
22

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Ir-1
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2/3”“
F BABUULAL BHAVARLAL JAIN
14.
15.
17.
18.
19.
polyoxyl 40 stearate, polysorbate 20, polysorbate 40, polysorbate 60, polysorbate
80, propylene glycol diacetate, propylene glycol monostearate, polyethylene
glycol, sodium lauryl sulfate, sodium stearate, sorbitan mono-laurate, sorbitan
mono-oleate, sorbitan mono-palmitate, sorbitanmonostearate, stearic acid, ethyl
alcohol, Propylene glycol, and emulsifying wax.
The method as claimed in claim 13, wherein the solubilizer comprises
Polyoxyethylenesorbitan fatty acid esters (1-15%), 1,2-Dihydroxypropane (1-10%),
a-Hydro-o-hydroxypoly(oxy-1,2-ethanediyl)(1-15%) and its one or the various
polymers.
The method as claimed in claim 1, wherein the emulsifier is present in the
composition in the range of about 0.2 to 1.0 % wt. of essential oil.
The method as claimed in claim 10, wherein the solubilizer is present in the
composition in the range of about 1 to 15.0 % wt. of essential oil.
The method‘ as claimed in claim 1, wherein the anti-oxidants comprises of
Rosmarinic acid (RA), carnosic acid, an ester of caffeic acid, 3,4
dihydroxyphenyllactic acid, tert-butyl-4-methoxyphenol, 2,6-di-tert-butyl-p-cresol,
p-cresol, alpha- bisabolol, carvacrol in the range of about 0.02 - 2 % of the
composition.
The method as claimed in claim 1,- wherein the ratio of contaminated water to
emulsion of the essential oil is in the range between 10:1 to 5000:1.
A composition for disinfecting water
microorganisms comprising 2
a)" an emulsifier; .
b) an essential oil composition selected from the group consisting of basil oils of
Ocimum sanctum; Ocimum basilicum; Ocimum grattissimum, Ocimum canum,
Ocimum citriodorum, Ocimum tenuiflorum L, Ocimum ciliatum, Ocimum
kilimandscharicum, Ocimum americanum; Ocimum campechianum, Ocimum
Viride; '
c) optionally a carrier oil comprising a plant oil;
d) optionally a natural anti-oxidant present in basil preserving the shelf life as well
as provide anti-oxidant effect; _
e) optionally containing an ingredient(s) enhances natural immunity.
contaminated with pathogenic
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Ir-ii
mi, :i .
:3;
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leer”
F BABUULAL BHAVARLAL JAIN
14.
15.
17.
18.
19.
polyoxyl 40 stearate, polysorbate 20, polysorbate 40, polysorbate 60, polysorbate
80, propylene glycol diacetate, propylene glycol monostearate, polyethylene
glycol, sodium lauryl sulfate, sodium stearate, sorbitan mono-laurate, sorbitan
mono-oleate, sorbitan mono-palmitate, sorbitanmonostearate, stearic acid, ethyl
alcohol, Propylene glycol, and emulsifying wax.
The method as claimed in claim 13, wherein the solubilizer comprises
Polyoxyethylenesorbitan fatty acid esters (1-15%), 1,2-Dihydroxypropane (1-10%),
a-Hydro-o-hydroxypoly(oxy-1,24ethanediyl)(1-15%) and its one or the various
polymers.
The method as claimed in claim 1, wherein the emulsifier is present in the
composition in the range of about 0.2 to 1.0 % wt. of essential oil.
The method as claimed in claim 10, wherein the solubilizer is present in the
composition in the range of about 1 to 15.0 % wt. of essential oil.
The method‘ as claimed in claim 1, wherein the anti-oxidants comprises of
Rosmarinic acid (RA), carnosic acid, an ester of caffeic acid, 3,4
dihydroxyphenyllactic acid, tert-butyl-4-methoxyphenol, 2,6-di-tert-butyl—p-cresol,
p-cresol, alpha- bisabolol, carvacrol in the range of about 0.02 - 2 % of the
composition.
The method as claimed in claim 1,- wherein the ratio of contaminated water to
emulsion of the essential oil is in the range between 10:1 to 5000:1.
A composition for disinfecting water
microorganisms comprising 2
a)" an emulsifier; .
b) an essential oil composition selected from the group consisting of basil oils of
Ocimum sanctum; Ocimum basilicum; Ocimum grattissimum, Ocimum canum,
Ocimum citriodorum, Ocimum tenuiflorum L, Ocimum ciliatum, Ocimum
kilimandscharicum, Ocimum americanum; Ocimum campechianum, Ocimum
viride; '
c) optionally a carrier oil comprising a plant oil;
d) optionally a natural anti-oxidant present in basil preserving the shelf life as well
as provide anti-oxidant effect; _
e) optionally containing an ingredient(s) enhances natural immunity.
contaminated with pathogenic
Dated this 29*“ day of December, 2014
6/ W
' 7’ SURESHKUMAR BABUULAL JAIN
_._ —,«. _».- --1 -.—.- 1:’ .-.- -.—.- ‘ ;— ;~-— 2- -4 ~~ ,-»
« - r - - - ‘ - L. .; ..~‘ ‘._ .; s_.
. ,2; .-' J. -5
23 27-Jan-2015I1718/267-MUM-2015IForm
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Ir-ii
mi, :i .
:3;
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leer”
F BABUULAL BHAVARLAL JAIN
14.
15.
17.
18.
19.
polyoxyl 40 stearate, polysorbate 20, polysorbate 40, polysorbate 60, polysorbate
80, propylene glycol diacetate, propylene glycol monostearate, polyethylene
glycol, sodium lauryl sulfate, sodium stearate, sorbitan mono-laurate, sorbitan
mono-oleate, sorbitan mono-palmitate, sorbitanmonostearate, stearic acid, ethyl
alcohol, Propylene glycol, and emulsifying wax.
The method as claimed in claim 13, wherein the solubilizer comprises
Polyoxyethylenesorbitan fatty acid esters (1-15%), 1,2-Dihydroxypropane (1-10%),
a-Hydro-o-hydroxypoly(oxy-1,24ethanediyl)(1-15%) and its one or the various
polymers.
The method as claimed in claim 1, wherein the emulsifier is present in the
composition in the range of about 0.2 to 1.0 % wt. of essential oil.
The method as claimed in claim 10, wherein the solubilizer is present in the
composition in the range of about 1 to 15.0 % wt. of essential oil.
The method‘ as claimed in claim 1, wherein the anti-oxidants comprises of
Rosmarinic acid (RA), carnosic acid, an ester of caffeic acid, 3,4
dihydroxyphenyllactic acid, tert-butyl-4-methoxyphenol, 2,6-di-tert-butyl—p-cresol,
p-cresol, alpha- bisabolol, carvacrol in the range of about 0.02 - 2 % of the
composition.
The method as claimed in claim 1,- wherein the ratio of contaminated water to
emulsion of the essential oil is in the range between 10:1 to 5000:1.
A composition for disinfecting water
microorganisms comprising 2
a)" an emulsifier; .
b) an essential oil composition selected from the group consisting of basil oils of
Ocimum sanctum; Ocimum basilicum; Ocimum grattissimum, Ocimum canum,
Ocimum citriodorum, Ocimum tenuiflorum L, Ocimum ciliatum, Ocimum
kilimandscharicum, Ocimum americanum; Ocimum campechianum, Ocimum
viride; '
c) optionally a carrier oil comprising a plant oil;
d) optionally a natural anti-oxidant present in basil preserving the shelf life as well
as provide anti-oxidant effect; _
e) optionally containing an ingredient(s) enhances natural immunity.
contaminated with pathogenic