DESC:CROSS- REFERENCE TO RELATED APPLICATION
This application claims the priority of the provisional application with serial number 202241071286 filed on 09/06/2023 titled as "A PRESERVATIVE COMPOSITION DERIVED FROM NATURAL COMPONENTS AND A METHOD OF ITS PREPARATION FOR THE IMPROVEMENT AND/OR STABILIZATION OF PERISHABLE COMPOUNDS" and the contents of which are incorporated in its entirety.

TECHNICAL FIELD OF THE INVENTION
The present invention relates to a polyherbal preservative composition and a method of its preparation for the improvement and/or stabilization of perishable skin care, cosmetics, wellness, pharmaceutical and food products. More particularly, the present invention relates to natural preservatives for improving the shelf-stability of skincare & haircare compositions, by antimicrobial and antioxidant actions of phytocomponents and moisture reducing natural salts.

BACKGROUND
The development of natural-based preservative formulations for personal care products such as shampoos and creams is critically needed in today's cosmetic industry. Traditional preservatives, including parabens, formaldehyde donors, and other synthetic chemicals, are linked to adverse health effects such as skin irritation, allergies, and potential carcinogenicity. Consequently, consumers increasingly prefer water-based cosmetics for their hydration benefits and smoother application. However, the high-water content in these formulations creates an ideal environment for microbial growth, leading to rapid spoilage and significantly reduced shelf life.
Extending the shelf life of water-based personal care products without compromising safety or efficacy is essential. While natural preservatives are favored for their perceived safety and environmental benefits, existing plant-based formulations often fall short. Many natural preservatives are only effective at certain pH levels, limiting their applicability and potentially causing skin irritation. Additionally, current extraction methods for botanical ingredients frequently fail to capture the full spectrum of bioactive compounds, resulting in inconsistent product quality and performance.
Maintaining an acidic pH is crucial for the efficacy and stability of these formulations. When the pH exceeds 7, active botanical ingredients and many chemical preservatives convert to their salt forms, reducing their antimicrobial efficacy and potentially causing skin irritation and product destabilization. This limitation underscores the necessity of keeping the pH on the acidic side to ensure effectiveness.
Moreover, many existing antimicrobial blends rely on quaternary compounds or halogenated molecules, particularly when parabens and formaldehyde donors are omitted. Although combinations of organic acids are sometimes used, their antimicrobial efficacy is limited to their acid form and low pH levels as they lose efficacy when converted to the salt form.
The current landscape of preservatives includes patents for various uses, such as US6361812 patent discloses a preservative system for use in foods, beverages, health care products, personal care products, comprising one or more isothiocyanates and an artificial preservative selected from sorbate preservatives, benzoate preservatives, or their mixtures. US20140322147A1 describes a certified organic ingredient used as a broad-spectrum preservative and disinfectant derived entirely from organic citrus fruit. US2010/058467 and US2010/058464 disclose plant extracts for oral care, while WO 2016/123716 A1 details a plant formulation for disinfecting and sanitizing surfaces. WO2013149323A1 and US7442391B2 disclose cosmetic skin care formulations with antimicrobial properties aimed at improving skin hygiene rather than preserving the formulations. Additionally, Indian Patent No. 311694 discloses an antimicrobial cosmetic composition that is entirely not plant-based. Hence, most of these patents focus on individual antimicrobial, antioxidant, or moisture-reducing actions without addressing the combined preservation effects of Phytocomponents on personal care formulations.
Given these challenges, there is a critical need for innovative natural-based preservative systems that can maintain the antimicrobial efficacy, stability, and safety of personal care products. Such advancements would meet the rising consumer demand for natural and safe cosmetics while ensuring the long-term viability and quality of water-based formulations. The present invention aims to develop methods to prevent microbial growth and extend shelf lives without artificial preservatives, enabling the creation of safe, organic, and natural products. This approach promises to set new standards in the cosmetic industry by combining the benefits of natural ingredients with the stringent performance standards required for modern personal care products.
The above-mentioned shortcomings, disadvantages and problems are addressed herein and which will be understood by reading and studying the following specification.

OBJECTIVE OF THE INVENTION
A principal object of the invention is to provide a Chemical free preservative composition for perishable products, with no harmful side effects to the user.
Another object of the invention is to provide an effective preservative composition for perishable products that contain antimicrobial or antioxidant or moisture-resistant actions of Phyto-components.
Yet another object of the invention is to provide a preservative composition that greatly increases the shelf life/expiration of the perishable product.
Yet another object of the invention is to provide a preservative composition that is stable at a wide range of room temperatures.
Yet another object of the invention is to provide a preservative composition that can be used at all temperature variations so that the shelf life of the perishable product is increased.
Yet another object of the invention is to make use of the antimicrobial and antioxidant properties of the natural Phyto components in combination with the dehydrating property of salts to bring about a natural preservative composition that is safe for oral as well as topical use in perishable products.
Yet another object of the invention is to use natural alternate sources of naturally available Phyto components as there is Limited availability of the standard most often used Natural preservative materials.
Yet another object of the invention is to determine whether the combination of the different components in the preservative composition shows an enhanced effect on the preservative property of the product versus when used alone.
Yet another object of the invention is to provide a Cost-effective preservative composition for use in perishable products
Yet another object of the invention is to provide a simple method to prepare a preservative composition using different Phyto component combinations
Yet another object of the invention is to provide a preservative composition for perishable products which is in ready-to-use form and does not require the further addition of any water or other component/s.
Yet another object of the invention is to provide a safe preservative composition that can be used universally in any perishable product to increase its shelf life.
Yet another object of the invention is to provide a preservative composition for perishable products which can be used immediately and does not alter the nature/quality of the existing perishable product.
Yet another object of the invention is to provide a preservative composition for perishable products which is eco-friendly and does not lead to water pollution.
Yet another object of the invention is to provide a good storage system for raw herbs to prevent oxidation, microbial action & moisturization during handling and storage.
These and other objectives and advantages of the embodiments herein will become readily apparent from the following detailed description taken in conjunction with the accompanying drawings.

SUMMARY OF THE INVENTION
The various embodiments of the invention provide a polyherbal preservative composition for the improvement and/or stabilization of perishable products using naturally occurring Phyto components. The formulation comprises a homogenous mixture of phyto-components, such as Curcumin, Eugenol, Linalool, Carnosic Acid and salts such as Sodium Chloride or Sodium Bicarbonate in a fixed ratio. The formulation is obtained by mixing the distillates of turmeric (Curcuma spp.), Clove (Syzygium spp.), Ginger (Zingiber spp.), Lavender (Lavandula spp.), Basil (Ocimum spp.), Rose (Rosa spp.), and Rosemary (Rosmarinus spp.) either individually or in combination to salts such as Sodium Chloride or Sodium Bicarbonate in a fixed ratio.
According to an embodiment of the present invention, the polyherbal preservative composition comprises Phyto-components of natural ingredients which are classified as Monoterpenoids: Antioxidants (AO): AOC, (Curcumin), AOE (Eugenol), AOL (Linalool); Antimicrobial Compounds (AMPC):AMPC1 (Carnosic Acid), AMPC 2 (Eugenol), AMPC 3 (Linalool); and Salts: SSC1 (Sodium Chloride), SSBC1 (Sodium bicarbonate) and mixed in a fixed ratio.
According to another embodiment of the present invention, a method of preparing a polyherbal preservative composition for the improvement and/or stabilization of perishable products using naturally occurring Phyto components is provided. The method comprises storage of the herbs collected, extraction of the phyto-components from the herbal ingredients and mixing them in a fixed proportion to get the final mixture of preservative composition. The method involves taking 10 g of the cleaned and freeze-dried stored herbs either individually [ Turmeric (Curcuma spp.), Clove (Syzygium spp.), Ginger (Zingiber spp.), Lavender (Lavandula spp.), Basil (Ocimum spp.), Rose (Rosa spp.), and Rosemary (Rosmarinus spp.)] or in combination (Turmeric, Rosemary, an admixture of Clove, Basil and Ginger, and an admixture of Basil, Rose and Lavender), boiling them in 60 ml of water, and collecting the distillates. The distillate extracts obtained are then mixed in equal proportion to obtain a homogenous mixture which is then mixed with the respective salt (either Sodium Chloride or Sodium bicarbonate) in a fixed ratio to obtain the preservative formulation.
According to another embodiment of the present invention, a method of preparing a polyherbal preservative composition involves obtaining a homogenous mixture by mixing the distillate of extracts from Turmeric (AOC-Curcumin), a distillate of mixed extracts of Clove, Basil, Ginger (AOE-Eugenol, AMPC2-Eugenol), a distillate of Rosemary (AMPC1-Carnosic Acid) and a distillate of mixed extracts of Basil, Rose, Lavender (AOL-Linalool, AMPC3-Linalool), in equal proportion i.e., in the ratio of 1:1:1:1 to a salt Sodium Chloride (SSC1-0.5% by weight).
These and other aspects of the embodiments herein will be better appreciated and understood when considered in conjunction with the following description and the accompanying drawings. It should be understood, however, that the following descriptions, while indicating preferred embodiments and numerous specific details thereof, are given by way of illustration and not of limitation. Many changes and modifications may be made within the scope of the embodiments herein without departing from the spirit thereof, and the embodiments herein include all such modifications.

BRIEF DESCRIPTION OF DRAWINGS
The other objectives, features, and advantages will occur to those skilled in the art from the following description of the preferred embodiment and the accompanying drawings in which:
FIG. 1 is a schematic showing the steps involved in the preparation of a preservative composition derived from natural components and a method of its preparation for the improvement and/or stabilization of perishable compounds, according to an embodiment of the present invention.

DETAILED DESCRIPTION OF THE INVENTION
In the following detailed description, a reference is made to the accompanying drawings that form a part hereof, and in which the specific embodiments that may be practiced is shown by way of illustration. The embodiments are described in sufficient detail to enable those skilled in the art to practice the embodiments and it is to be understood that the logical, mechanical, electronic and other changes may be made without departing from the scope of the embodiments. The following detailed description is therefore not to be taken in a limiting sense.
According to an embodiment of the present invention, the formulation comprises a homogenous mixture obtained from mixing a distillate containing an aqueous extract from the rhizome of Curcuma spp. having the antimicrobial and antioxidant phytocomponent Curcumin; a distillate containing an aqueous extract from the aerial plant parts of Rosmarinus spp. having the antimicrobial and antioxidant phytocomponent Carnosic Acid; a distillate containing an ‘polyherbal aqueous extract’ obtained from the dried buds of Syzygium spp., the aerial plant parts of Ocimum spp. and the rhizome of Zingiber spp., having the the antimicrobial and antioxidant phytocomponent Eugenol; a distillate containing a ‘polyherbal aqueous extract’ obtained from the aerial plant parts of Ocimum spp., aqueous extract from floral parts of Rosa spp. and aqueous extract from the floral parts of Lavandula spp., containing the antimicrobial and antioxidant phytocomponent Linalool; a salt selected from a group comprising Sodium chloride, Sodium Bicarbonate, etc.; and wherein the salt component is present as 0.5 % by weight; and Water or oil; wherein the distillate of the aqueous extract from the rhizome of Curcuma spp. is in the range of 0.5-1% by weight, wherein the distillate of the aqueous extract from the aerial plant parts of Rosmarinus spp. is 1% by weight, wherein the polyherbal distillate obtained from the dried buds of Syzygium spp., the aerial plant parts of Ocimum spp. and the rhizome of Zingiber spp. are in the range of 0.5-2% by weight, and wherein the polyherbal distillate obtained from the aerial plant parts of Ocimum spp. , aqueous extract from the floral parts of Rosa spp., the floral parts of Lavandula spp. are present in the range between 0.5-1% by weight.
According to an embodiment of the present invention, the formulation comprises of a homogenous mixture obtained from mixing an antioxidant (Phenolic: curcumin, eugenol, linalool classified as monoterpenoids; also classified as Antioxidants (AO): AOC (Curcumin), AOE (Eugenol), AOL (Linalool) for the purpose of depicting in the tables and charts in the specification) an antimicrobial (Carnosic Acid, Linalool, Eugenol classified as monoterpenoids; also classified as Antimicrobial Compounds (AMPC): AMPC1 (Carnosic Acid), AMPC 2 (Eugenol), AMPC 3 (Linalool) for the purpose of depicting in the tables and charts in the specification) and salt (Sodium Chloride SSC1, Sodium Bicarbonate SSBC1).
FIG. 1 shows a schematic showing the steps involved in the preparation of a polyherbal preservative composition to stabilize perishable personal care products according to an embodiment of the present invention. With respect to FIG.1, the method comprises selection and storage of the said herbs collected, extraction of the phytocomponents from the herbal ingredients and mixing them to get the final mixture of the preservative. The first step in the method involves the selection and preparation preparation of the herbs wherein the said herbs are selected from a group comprising Curcuma spp., Rosmarinus spp., Syzygium spp., Ocimum spp., Zingiber spp., Lavandula spp. and Rosa spp. [291]. The rhizomes of turmeric (Curcuma longa) are cleaned, dried under shade for 2-3 days, subjected to drying by sublimation of moisture through freeze-drying, stored under liquid nitrogen [291] and 10 g of this frozen stored herb is taken and boiled with 60 ml of water and the distillate is collected [292]. The aerial parts of rosemary (Rosmarinus officinalis) are taken, cleaned, and dried under shade for 2-3 days, subjected to drying by sublimation of moisture through freeze-drying, stored under liquid nitrogen [291] and 10 g of this frozen stored herb is taken and boiled with 60 ml of water and the distillate is collected. [292]. The dried buds of clove (Syzygium aromaticum), aerial parts of Basil (Ocimum basilicum) and rhizomes of ginger (Zingiber officinale) are taken, cleaned, and dried under shade for 2-3 days, subjected to drying by sublimation of moisture through freeze-drying, stored under liquid nitrogen [291] and 10 g of this frozen stored admixture of herbs is taken and boiled with 60 ml of water and the distillate is collected [292]. The aerial parts of Basil (Ocimum basilicum), floral parts of rose (Rosa rubiginosa) and floral parts of lavender (Lavandula spp.) are taken, cleaned, and dried under shade for 2-3 days, subjected to drying by sublimation of moisture through freeze-drying, stored under liquid nitrogen [291] and 10 g of this frozen stored admixture of herbs is taken and boiled with 60 ml of water and the distillate is collected [292]. The distillate extracts of the herbs either individually or in specific combinations are mixed in different proportions to obtain a homogenous ‘mixture’ [293] which is then added to the salt selected from a group comprising Sodium chloride, Sodium Bicarbonate, etc. 0.5% by weight to obtain the preservative mixture/formulation [294] which was then subjected to shelf-life studies [295].
The crude herbs were procured from a reputed supplier and they were then subjected to an extraction process to get the desired phytocomponents used in the experimentation. Alternatively, the readymade commercially available plant extracts of the phytocomponents mentioned above, were also used.
According to another embodiment of the present invention, the polyherbal preservative composition comprises Phyto-components of natural ingredients which are classified as: Antioxidants (AO): AOC (Curcumin), AOE (Eugenol), AOL (Linalool); Antimicrobial Compounds (AMPC): AMPC1 (Carnosic Acid), AMPC 2 (Eugenol), AMPC 3 (Linalool); and Salts as desiccants: SSC1 (Sodium Chloride), SSBC1 (Sodium bicarbonate).
According to another embodiment of the present invention, the polyherbal preservative composition comprises Phyto-components of natural ingredients which behave as both Antioxidants (AO) and antimicrobial Compounds (AMPC): Eugenol-AOE, AMPC2, Linalool-AOL, AMPC3.
Since water based personal care products are preferred for usage, the major problem faced is that this water serves as the most supporting environment for the microbes to grow. Hence, according to another embodiment of the present invention, the SSC1 (Sodium Chloride), SSBC1 (Sodium bicarbonate) used absorb moisture, disturbing the integrity of the cell membranes of the microbes thereby also showing antimicrobial activity in addition to them being desiccants.

EXPERIMENTAL DETAILS
The effectiveness of the formulation is established using the MIC test (Microbial Inhibitory Composition using agar well Diffusion method) and ORAC (The Oxygen Radical Absorbance Capacity assay method based on the level of fluorescence emitted) values.
Table 1: Tests Taken Up to study the efficacy of the preservative:
Name of Test Preferred Range Method
MIC - Minimum Inhibitory Concentration 1 to 20,000+ Broth Dilution Method
ORAC - Oxygen Radical Absorbance Capacity 0.3 to 1 Lakh+ ORAC Assay
pH 4.5 to 7 pH meter is used
Viscosity 4000 to 50,000 cps
Viscometer is used
Microbiological Testing No growth to be found Aerobic plate count
Centrifugation No Phase separation Differential centrifugation
Thermal Stability No Phase separation Differential Thermal Analysis
TFM - Total Fatty Matter 3 to 10% Titration Method
Foam Height 120 to 190 ml Ross-Mile’s method

According to another embodiment of the present invention, the efficacy of these formulations was ascertained by performing the Shelf-life tests on the various types of topical personal care formulations viz, a face cream formulation, a face wash, a shampoo formulation, a conditioner formulation, a body lotion formulation and Hygienic Intimate wash formulation that were preserved with 0.5 to 2.0 % of the Preservative Formulation as exemplified below in the experimental section.

Table 2: Shelf-life test to determine % of Preservative Formulation to be used in the personal care product

% of Preservative Formulation used Observation
0.5% Product was stable for only a period of 9-10 Months
1% Product was stable for 2 years
2% Product was stable for 3+ years

2% preservative formulation, though showed a promise of stability of over 3+ years, was not considered because of the possibility of skin irritations at higher concentration of phytocomponents used. Therefore, the concentration of 1% considered safe for human skin was used in subsequent testing protocols.
The appropriate concentration of the phytocomponent preservative formulation in the present invention varies depending on the personal care product formulation and its water content, which ranges from 21% to 66% by weight of the total topical Preservative formulation. The antimicrobial efficacy of these preservative formulations was tested across various formulations and against different microbes to determine their effectiveness and the promising resultant formulations were further evaluated for their Shelf-life stability through further tests.
The extraction of phytocomponents was done through the distillation process wherein, a heat pump was used which was set at 80? along with vacuum pressure of 600 mm of Hg. This pressure helps in reducing the boiling point to 65? and increasing the extract obtained.

Preparation of different Preservative Formulation compositions

Table 3: Phytocomponents used in the experimentation:
Activity Phytocomponent Code % Range used

Anti-Oxidants Curcumin (Curcuma spp.) AOC 0.5-1%
Eugenol (Syzygium spp.,Ocimum spp., Zingiber spp.) AOE 0.5-2%
Linalool (Ocimum spp., Rosa spp., Lavandula spp.) AOL 0.5-1%

Anti-Microbial Carnosic acid (Rosmarinus spp.) AMPC1 1%
Eugenol (Syzygium spp.,Ocimum spp., Zingiber spp.) AMPC2 0.5-2%
Linalool (Ocimum spp., Rosa spp., Lavandula spp.) AMPC3 0.5-1%
Salts Sodium Chloride SSC1 0.5%
Sodium Bicarbonate SSBC1 0.5%

Preservative Formulation 1- 1% of AOE, AOC, AMPC1, AMPC3 each along with 0.5% of SSC1 was mixed thoroughly and subjected to MIC and ORAC testing. Based on the testing results, the sample was shortlisted to be used in the final product which was then subjected to accelerated stability testing.
Preservative Formulation 2 - 1% of AOL, AOC, AMPC2, AMPC1 each along with 0.5% of SSBC1 was mixed thoroughly and subjected to MIC and ORAC testing. Based on the testing results, the sample was shortlisted to be used in the final product which was then subjected to accelerated stability testing.
Preservative Formulation 3- 0.5% of AOC, 1% of AOE, AMPC2, AMPC3 each along with 0.5% of SSC1 was mixed thoroughly and subjected to MIC and ORAC testing. Based on the testing results, the sample was shortlisted to be used in the final product which was then subjected to accelerated stability testing.
Preservative Formulation 4 - 0.5% of AOL, 1% of AOC, AMPC2, AMPC1 each along with 0.5% of SSBC1 was mixed thoroughly and subjected to MIC and ORAC testing. Based on the testing results, the sample was shortlisted to be used in the final product which was then subjected to accelerated stability testing.

Testing of the Preservative Formulation compositions for antimicrobial activity:
In one embodiment of the invention, the Preservative Formulation 1 comprising AOE+ AOC+AMPC 1+ AMPC 3+ SSC1 in the ratio of 1:1:1:0.5% (by weight) is added to a shampoo base and the ORAC value obtained was 4401.6 mmol and MIC for Gram Positive Bacteria was 6.93 mg/ml; Gram Negative Bacteria was 18.18 mg/ml; Fungi was 12 mg/ml.
In another embodiment of the invention, the Preservative Formulation 2 comprising AOL+ AOC+AMPC 1+ AMPC 2+ SSBC1 in the ratio of 1:1:1:0.5% (by weight) is added to a shampoo base and the ORAC value obtained was 4377.55 mmol and MIC for Gram Positive Bacteria was 12479.25 mg/ml; Gram Negative Bacteria was 32 mg/ml; Fungi was 7.25 mg/ml.
In another embodiment of the invention, the Preservative Formulation 3 comprising AOC + AOE+AMPC 2+ AMPC 3+ SSC1 in the ratio of 0.5:1:1:0.5% (by weight) is added to a Body Lotion and the ORAC value obtained was 4401.68 mmmol and MIC for Gram Positive Bacteria was 12477.68 mg/ml; Gram Negative Bacteria was 19.25 mg/ml; Fungi was 5.25 mg/ml.
In another embodiment of the invention, the Preservative Formulation 4 comprising AOL+ AOC+AMPC 1+ AMPC 2+ SSBC1 in the ratio of 0.5:1:1:0.5% (by weight) is added to a Body Lotion and the ORAC value obtained was 4377.5 mmol and MIC for Gram Positive Bacteria was 12475 mg/ml; Gram Negative Bacteria was 32 mg/ml; Fungi was 7.25 mg/ml.

Table 4: Preservative formulations efficacy testing for antimicrobial activity
Product Name Preservative formulations Phyto Component Composition (%) ORAC MIC
Shampoo/ Face Wash/ Hygienic Intimate Wash 1 AOE 1% 4401.6 mmol Gram Positive - 6.93 mg/ml
Gram Negative - 18.18 mg/ml
Fungi - 12.85 mg/ml
AOC 1%
AMPC1 1%
AMPC3 1%
SSC1 0.5%
2 AOL 1% 4377.55 mmol
Gram Positive - 6.3 mg/ml
Gram Negative - 12479.25 mg/ml
Fungi - 32 mg/ml
AOC 1%
AMPC1 1%
AMPC2 1%
SSBC1 0.5%
Body Lotion/ Face Cream/ Conditioner 3 AOC 0.5% 4401.68 mmol Gram Positive - 12477.68 mg/ml
Gram Negative - 19.25 mg/ml
Fungi -5.25 mg/ml
AOE 1%
AMPC2 1%
AMPC3 1%
SSC1 0.5%
4 AOL 0.5% 4377.58 mmol Gram Positive - 12475 mg/ml
Gram Negative - 32 mg/ml
Fungi - 7.25 mg/ml
AOC 1%
AMPC2 1%
AMPC1 1%
SSBC1 0.5%

Preservative Formulation antimicrobial activity test results:
The above results in Table 4 show that the Preservative Formulation 1 has more capacity to act as a preservative as it showed Higher ORAC and lower MIC values.
The ORAC is 1% better in Preservative Formulation 1 than Preservative Formulation 2. The MIC of Gram-Positive bacteria is 0.9% higher in Preservative Formulation 1 than Preservative Formulation 2. The MIC of Gram-Negative bacteria is 685%+ higher in Preservative Formulation 2 than Preservative Formulation 1. The MIC of Fungi is 149% higher in Preservative Formulation 2 than Preservative Formulation 1. Hence, as there is a decreased MIC in Preservative Formulation 1 when compared to Preservative Formulation 2, the Preservative Formulation 1 was considered for the further study protocol.
Further Preservative Formulation 3 was compared with Preservative Formulation 4 based on percent weight ratio as shown in Table 4.
Preservative Formulation 3 has more capacity to act as a preservative as it has higher ORAC and lower MIC values. This helps in protecting the product for a longer period.
The ORAC is 1% better in Preservative Formulation 3 than Preservative Formulation 4. The MIC of Gram-Positive bacteria is 0.02% higher in Preservative Formulation 3 than Preservative Formulation 4. The MIC of Gram-Negative bacteria is 40% higher in Sample 4 than Sample 3. The MIC of Fungi is 27.5% higher in Preservative Formulation 4 than Preservative Formulation 3. Hence, Preservative Formulation 3 having a good antifungal property was considered for the further study protocol.

Methodology of the Tests Taken Up in the study:
1. MIC - MIC determined by microbroth dilution method. Serial concentrations of phytocomponents were prepared using Mueller-Hinton broth as diluent and inoculated with 20µl of bacterial inoculums for 24H at 37? for the growth of bacteria. The lowest concentration at which there was no growth was taken as MIC of the phytocomponent.
o Control: Gram Negative – Cefepime
o Gram Positive – Erythromycin
o Fungi - Clotrimazole
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7913839/
2. ORAC - Experimental Procedure
Fluorescein solution (10 nM), Trolox standards (800 µM – 25 µM, 2x serial dilution) and AAPH solution (240mM) were prepared in PBS buffer pH 7.4.
Plate preparation: To each active well was added 25 µL of either buffer (for the blank) or Trolox standards. The plate was placed into the plate reader with incubation at 37 °C, and the experiment for the determination of the antioxidant capacity was started.
3. Determination of pH: The pH value of a liquid can be determined potentiometrically by means of the glass electrode, a reference electrode and a pH meter either of the digital or analogue type.
Viscosity: Viscosity is a property of a liquid, which is closely related to the resistance to flow.
The liquid under test is filled in a U tube viscometer in accordance with the expected viscosity of the liquid so that the fluid level stands within 0.2 mm of the filling mark of the viscometer when the capillary is vertical and the specified temperature is attained by the test liquid. The liquid is sucked or blown to the specified weight of the viscometer and the time taken for the meniscus to pass the two specified marks is measured. The kinematic viscosity in centistokes is calculated from the following equation: Kinematic viscosity = kt Where k = the constant of the viscometer tube determined by observation on liquids of known kinematic viscosity; t = time in seconds for meniscus to pass through the two specified marks.
5. Centrifugation: Centrifuge tube to be filled with the sample and placed in the centrifuge apparatus by maintaining the balance. Secure the lids of the tubes tightly. Close the lid of the apparatus and set the run speed and time. Post the time period, the tubes should be removed and checked for any phase separation.

6. Thermal stability: With the help of spatula, insert the cream into a glass bottle and tap it to settle to the bottom. Fill up to two third capacity of bottle and insert plug and tighten the cap. Keep the filled bottle erect inside the incubator at 45+ 1 0C for 48 h. The sample was taken to have passed the test, as there was no phase separation observed.

7. Microbiological Tests:
o E. coli - Place the prescribed quantity in a sterile screw-capped container, add 50 ml of nutrient broth, shake, allow to stand for 1 hour (4 hours for gelatin) and shake again. Loosen the cap and incubate at 37? for 18 to 24 hours.
By means of an inoculating loop, strip a portion from the enrichment culture (obtained in the previous test) on the surface of MacConkey agar medium. Cover and invert the dishes and incubate. Upon examination, if none of the colonies are brick-red in colour and have a surrounding zone of precipitated bile, the sample meets the requirements of the test for the absence of Escherichia coli.
o Salmonella - 1 g or 1 ml of the product to 100 ml of nutrient broth in a sterile screw-capped jar, shake, allow to stand for 4 hours and shake again. Loosen the cap and incubate at 35? to 37? for 24 hours.
Primary test: Add 1.0 ml of the enrichment culture to each of the two tubes containing (a) 10 ml of selenite F broth and (b) tetrathionate-bile-brilliant green broth and incubate at 36? to 38? for 48 hours. From each of these two cultures, the following agar media: bismuth sulphite agar, brilliant green agar. Incubate the plates at 36? to 38? for 18 to 24 hours. Upon examination, the sample meets the requirements of the test for the absence of the genus Salmonella.
o Pseudomonas aeruginosa - inoculate 100 ml of fluid soyabean-casein digest medium with a quantity of the solution, suspension or emulsion thus obtained containing 1 g or 1 ml of the preparation being examined. Mix and incubate at 35? to 37? for 24 to 48 hours. Examine the medium for growth and if growth is present, streak a portion of the medium on the surface of cetrimide agar medium, each plated on Petri dishes. Cover and incubate at 35? to 37? for 18 to 24 hours.
o Staphylococcus aureus – Procedure similar to Pseudomonas aeruginosa. Upon examination of the incubated plates, the sample meets the requirements for the absence of Staphylococcus aureus if colonies are seen.
According to an embodiment of the present invention, to study the efficacy of the naturally derived formulations, the shelf- stability of perishable personal care products is evaluated as per the norms of ICH - International Conference on Harmonization and API - Ayurvedic Pharmacopoeia of India.

SAMPLE PREPARATION FOR STABILITY TESTING:

1% of the Preservative Formulation mentioned above is added to the base (i.e., the products like Shampoo, Face Wash, Hygienic Intimate wash, Body lotion, Face cream and Conditioner) making the total composition to 100% (personal care product base + Preservative Formulation) and this was tested for the 6 months accelerated stability study, which proved the potency of the Preservative Formulation and the results have been positive.

Table 5. Shelf-life Stability Testing results of the polyherbal preservative formulations.
Product Name Preservative Sample Accelerated Test Conditions Accelerated Test Parameters Test Frequency Observation Accelerated test duration
Shampoo/ Face Wash/ Hygienic Intimate Wash
Sample 1 Temperature - 40°C ± 2°C
Relative humidity - 75% RH ± 5% RH Appearance, Colour, Odour, Texture, pH, Foam Height, Microbiological Study etc as per API standard 0th Day, 30th Day, 60th Day, 90th Day, 180th Day The results match the criteria mentioned in API and ICH guidelines 6 Months

Body Lotion/ Face Cream/ Conditioner Sample 3 Temperature - 40°C ± 2°C
Relative humidity - 75% RH ± 5% RH Appearance, Colour, Odour, Texture, pH, Total Fatty Matter, Total Residue Content, Microbiological Study 0th Day, 30th Day, 60th Day, 90th Day, 180th Day The results match the criteria mentioned in API 6 Months

*As per ICH Guidelines, the products were stable for a period of 6 months, which confirms that the shelf life of the product to be 2 Years
In an embodiment of the invention, the preservative composition added to a shampoo base was tested for the Expected Shelf Life at Accelerated Test Conditions of Temperature of 40°C, Relative humidity - 75% RH on Test Parameters such as Appearance, Colour, Odour and Texture at accelerated test duration of 6 months (as per ICH Guidelines, accelerated stability study can give a 24 Months shelf life to the product) and the results matched the criteria mentioned in API (Ayurvedic Pharmacopoeia of India).
In addition, it has been determined that the compositions of the herein disclosed personal care products will preferably have a pH value in a range from about 3 to about 7, more preferably from about 4 to about 6, and most preferably from about 4.2 to about 5.9.
Based on the antimicrobial test studies, Preservative Formulation 1 (comprising of 1% of AOE, AOC, AMPC3, AMPC1 each along with 0.5% of SSC1) and Preservative Formulation 3 (comprising of 0.5% of AOC, 1% of AOE, AMPC2, AMPC3 each along with 0.5% of SSC1) were considered for the further study protocol including shelf-life efficacy studies.
According to an embodiment of the present invention 6 months accelerated stability test of the personal care products such as Shampoo, Face Wash, Hygienic Intimate wash, Body lotion, Face cream and Conditioner, as per ICH guidelines was conducted using the chosen compositions -Preservative Formulation 1 and Preservative Formulation 3 to establish the shelf-life of the final products. Parameters tested include appearance, color, odour, texture, pH, Foam height, Non-volatile alcohol soluble matter, viscosity, total yeast and mould count, total plate count, and presence of microbes like E. coli, Pseudomonas aeruginosa, Staphylococcus aureus and Salmonella.
According to an embodiment of the present invention, the final composition of the product contains 1% of the Preservative Formulation 1 (comprising of 1% ginger rhizome distillate-AOC, 1% distillate of Rosemarinus spp.- AMPC1, 1% of ‘polyherbal distillate mixture’ of Syzygium spp., Ocimum spp., and Zingiber spp. -AOE, polyherbal distillate mixture’ of Ocimum spp., Lavandula spp. and Rosa spp -AMPC3, each along with 0.5% of Sodium Chloride-SSC1) about 42-78% of Personal care product base, having 21-57% water, making the total composition to 100% with pH 4.2 to 5.5.
Table 6: Formula of the polyherbal preservative composition in the personal care product (Shampoo/ Face Wash, Hygienic Intimate Wash): Example 1
Component %
Preservative Formulation 1 1%
Aqua (Demineralized/ Distilled Water) 21% - 57%
Other Botanicals 20% - 50%
Foaming Agents 11% - 21%
Moisturizing Agents 4% - 14%
Chelating + Thickening Agent + Essential Oils 0.6% - 2.8%
pH (0th Day) 4.2 - 5.5

According to an embodiment of the present invention,the final composition of the product contains 1% of the Preservative Formulation 3 (comprising of 0.5% of ginger rhizome distillate-AOC, 2% of ‘polyherbal distillate mixture’ of Syzygium spp., Ocimum spp., and Zingiber spp.-1% AOE + 1% AMPC2, and 1% ‘polyherbal distillate mixture’ of Ocimum spp., Rosa spp. and Lavandula spp.-AMPC3 along with 0.5% of Sodium Chloride-SSC1) about 31-53 % of Personal care product base, having 46-68% water, making the total composition to 100% with pH 4.3 to 5.0

Table 7: Formula of the polyherbal preservative composition in the personal care product (Body Lotion/ Conditioner/ Face Cream): Example 2
Component %
Preservative Formulation 3 1%
Aqua (Demineralised/ Distilled Water) 46% - 68%
Other Botanicals 11% - 23%
Moisturizing + Conditioning Agents 14% - 29%
Chelating + Thickening Agent + Essential Oils 1% - 1.7%
pH (0th Day) 4.3 to 5.0

The working of the said composition is illustrated by the following non limiting illustrative examples.

Example 1: Shampoo with the Preservative Formulation- Table 8.
S. No. Test Parameter Method 0th Day 30th Day 60th Day 90th Day 180th Day
1 Appearance API Dark Brown coloured Shampoo Dark Brown coloured Shampoo Dark Brown coloured Shampoo Dark Brown coloured Shampoo Dark Brown coloured Shampoo
2 Colour As per Standard Method Dark Brown Dark Brown Dark Brown Dark Brown Dark Brown
3 Odour As per Standard Method Characteristic Characteristic Characteristic Characteristic Characteristic
4 Texture As per Standard Method Liquid Liquid Liquid Liquid Liquid
5 pH IS 7884 4.821 4.825 4.827 4.875 4.96
6 Foam Height IS 7884 140 ml 142 ml 145 ml 146 ml 148 ml
7 Non volatile alcohol soluble matter IS 7884 15.62% 15.77% 15.83% 15.85% 16.24%
8 Viscosity API 3245 cps 3242 cps 3246 cps 3240 cps 3248 cps
9 Total Yeast & Mould count API <10 cfu/ g <10 cfu/ g <10 cfu/ g <10 cfu/ g <10 cfu/ g
10 Total plate count API <10 cfu/ g <10 cfu/ g <10 cfu/ g <10 cfu/ g <10 cfu/ g
11 E.Coli API Absent/ g Absent/ g Absent/ g Absent/ g Absent/ g
12 Pseudomonas aeruginosa API Absent/ g Absent/ g Absent/ g Absent/ g Absent/ g
13 Staphylococcus aureus API Absent/ g Absent/ g Absent/ g Absent/ g Absent/ g
14 Salmonella API Absent/ g Absent/ g Absent/ g Absent/ g Absent/ g

1% of the Preservative Formulation 1 (comprising of 1% of AOE, AOC, AMPC3, AMPC1 each along with 0.5% of SSC1) is added to the Shampoo base making the total composition to 100% and this was tested for the 6 months accelerated stability study, for various parameters. The color, odor, texture remained stable throughout the testing process. pH varied between 4.8 to 4.9 by the end of 6 months study and was within the standard limits. Foam Height varied between 140 ml to 148 ml and was within the standard limits. Viscosity varied between 3245 to 3248 which is within the standard limits. There was no microbial growth seen in the product by the end of the 6th month study period. Overall, there were no significant changes observed in the parameters tested at accelerated test duration of 6 months which, as per ICH Guidelines, can give a 24 Month i.e., 2-year shelf life to the product, thereby proving the potency of the Preservative Formulation-Table 8.

Example 2: Face wash with Preservative Formulation- Table 9
S. No. Test Parameter Method 0th Day 30th Day 60th Day 90th Day 180th Day
1 Appearance API Brick red coloured viscous liquid Brick red coloured viscous liquid Brick red coloured viscous liquid Brick red coloured viscous liquid Brick red coloured viscous liquid
2 Odour API Characteristic Characteristic Characteristic Characteristic Characteristic
3 Texture API Liquid Liquid Liquid Liquid Liquid
4 Colour API Brick Red Brick Red Brick Red Brick Red Brick Red
5 pH API 5.540 5.528 5.577 5.584 5.96
6 Spreadability API 8.2 cm 8.1 cm 8.3 cm 8.4 cm 8.6 cm
7 Viscosity API 6785 cps 6780 cps 6779 cps 6785 cps 6782 cps
8 Total Fatty Matter API 3.11% 3.15% 3.12% 3.13% 3.15%
9 Matter insoluble in alcohol API 4.22% 4.20% 4.24% 4.26% 4.3%
10 Foam Height API 170 ml 172 ml 175 ml 176 ml 182 ml
11 Total Yeast & Mould count API <10 cfu/g <10 cfu/g <10 cfu/g <10 cfu/g <10 cfu/g
12 Salmonella API Absent/g Absent/g Absent/g Absent/g Absent/g
13 E.Coli API Absent/g Absent/g Absent/g Absent/g Absent/g
14 Pseudomonas aeruginosa API Absent/g Absent/g Absent/g Absent/g Absent/g
15 Staphylococcus aureus API Absent/g Absent/g Absent/g Absent/g Absent/g
16 Total plate count API <10 cfu/g <10 cfu/g <10 cfu/g <10 cfu/g <10 cfu/g

1% of the Preservative Formulation 1 (comprising of 1% of AOE, AOC, AMPC3, AMPC1 each along with 0.5% of SSC1) is added to the Face wash base making the total composition to 100% and this was tested for the 6 months accelerated stability study. The color, odor, texture of the product was stable throughout the testing process. pH varied between 5.5 to 5.9 which is within the standard limits. Viscosity varied between 6779 to 6785, which is within the standard limits. Foam height varied from 170 ml to 182 ml, which is within the standard limits. There was no microbial growth seen in the product by the end of the 6th month study period. Overall, there were no significant changes observed in the parameters tested at accelerated test duration of 6 months which, as per ICH Guidelines, can give a 24 Month i.e., 2-year shelf life to the product, thereby proving the potency of the Preservative Formulation-Table 9.

Example 3: Intimate Hygiene wash with the Preservative Formulation- Table 10.
S. No. Test Parameter Method 0th Day 30th Day 60th Day 90th Day 180th Day
1 Appearance API Brick red coloured liquid Brick red coloured liquid Brick red coloured liquid Brick red coloured liquid Brick red coloured liquid
2 Odour API Characteristic Characteristic Characteristic Characteristic Characteristic
3 Texture API Liquid Liquid Liquid Liquid Liquid
4 Colour API Brick Red Brick Red Brick Red Brick Red Brick Red
5 pH API 4.201 4.208 4.211 4.252 4.201
6 Specific gravity API 1.02 1.02 1.02 1.02 1.034
7 Viscosity API 6788 cps 6782 cps 6788 cps 6785 cps 6789 cps
8 Total plate count API <10 cfu/g <10 cfu/g <10 cfu/g <10 cfu/g <10 cfu/g
9 Total Yeast & Mould count API <10 cfu/g <10 cfu/g <10 cfu/g <10 cfu/g <10 cfu/g
12 Salmonella API Absent/ml Absent/ml Absent/ml Absent/ml Absent/ml
13 E.Coli API Absent/ml Absent/ml Absent/ml Absent/ml Absent/ml
14 Pseudomonas aeruginosa API Absent/ml Absent/ml Absent/ml Absent/ml Absent/ml
15 Staphylococcus aureus API Absent/ml Absent/ml Absent/ml Absent/ml Absent/ml

In order to maintain the optimum pH for this Hygienic Intimate Wash, Citric acid has been used.
1% of the Preservative Formulation 1 (comprising of 1% of AOE, AOC, AMPC3, AMPC1 each along with 0.5% of SSC1) is added to the Intimate Hygiene Wash base making the total composition to 100% and this was tested for the 6 months accelerated stability study. The color, odor, texture of the product was stable throughout the testing process. pH was within 4.2 which is within the standard limits. Viscosity varied between 6782 to 6788 which is within the standard limits. There was no microbial growth seen in the product by the end of the 6th month study period. Overall, there were no significant changes in the parameters tested at accelerated test duration of 6 months which, as per ICH Guidelines, can give a 24 Month i.e., 2-year shelf life to the product, thereby proves the potency of the Preservative Formulation-Table 10.

Example 5: Body Lotion with the Preservative Formulation- Table 11
S. No. Test Parameter Method 0th Day 30th Day 60th Day 90th Day 180th Day
1 Appearance API Brownish Yellow Colour Brownish Yellow Colour Brownish Yellow Colour Brownish Yellow Colour Brownish Yellow Colour
2 Odour As per Standard Method Characteristic Characteristic Characteristic Characteristic Characteristic
3 Texture As per Standard Method Semi solid Semi solid Semi solid Semi solid Semi solid
4 Colour As per Standard Method Brownish Yellow Brownish Yellow Brownish Yellow Brownish Yellow Brownish Yellow
5 pH As per Standard Method 4.346 4.351 4.346 4.346 4.346
6 Total Residue Content IS:6608 37.13% 37.14% 37.13% 37.13% 37.13%
7 Total Fatty Matter IS:6608 5.26% 5.25% 5.26% 5.26% 5.26%
8 Viscosity API 2890 cps 2890cps 2886 cps 2889 cps 2889 cps
9 Total Yeast & Mould count API <10 cfu/g <10 cfu/g <10 cfu/g <10 cfu/g <10 cfu/g
10 Salmonella API Absent/ g Absent/ g Absent/ g Absent/ g Absent/ g
11 E.Coli API Absent/ g Absent/ g Absent/ g Absent/ g Absent/ g
12 Pseudomonas aeruginosa API Absent/ g Absent/ g Absent/ g Absent/ g Absent/ g
13 Staphylococcus aureus API Absent/ g Absent/ g Absent/ g Absent/ g Absent/ g
14 Total plate count API <10 cfu/g <10 cfu/g <10 cfu/g <10 cfu/g <10 cfu/g

1% of the Preservative Formulation 3 (comprising of 0.5% of AOC, 1% of AOE, AMPC2, AMPC3 each along with 0.5% of SSC1) is added to the Body Lotion base making the total composition to 100% and this was tested for the 6 months accelerated stability. The color, odor, texture of the product was stable throughout the testing process. pH was within 4.2 which is within the standard limits. There was no microbial growth seen in the product by the end of the 6th month study period. Overall, there were no significant changes in the parameters tested at accelerated test duration of 6 months which, as per ICH Guidelines, can give a 24 Month i.e., 2-year shelf life to the product, thereby proving the potency of the Preservative Formulation-Table 11.
Example 6: Conditioner with the Preservative Formulation- Table 12
S. No. Test Parameter Method 0th Day 30th Day 60th Day 90th Day 180th Day
1 Appearance API Off white coloured cream Off white coloured cream Off white coloured cream Off white coloured cream Off white coloured cream
2 Colour API Off white Off white Off white Off white Off white
3 Odour As per Standard Method Characteristic Characteristic Characteristic Characteristic Characteristic
4 Texture As per Standard Method Semi solid Semi solid Semi solid Semi solid Semi solid
5 pH As per Standard Method 5.064 5.030 5.016 5.024 5.106
6 Rancidity IS 7679 Absent Absent Absent Absent Absent
7 Water Content IS 9740 67.55% 67.99% 67.63% 67.72% 67.36%
8 Thermal Stability IS 6608 Stable Stable Stable Stable Stable
9 Total Fatty matter IS 6608 6.14% 6.15% 6.22% 6.23% 6.15%
10 Total Yeast & Mould count API <10 cfu/g <10 cfu/g <10 cfu/g <10 cfu/g <10 cfu/g
11 Total plate count API <10 cfu/g <10 cfu/g <10 cfu/g <10 cfu/g <10 cfu/g
12 E.Coli API Absent/ g Absent/ g Absent/ g Absent/ g Absent/ g
13 Pseudomonas aeruginosa API Absent/ g Absent/ g Absent/ g Absent/ g Absent/ g
14 Staphylococcus aureus API Absent/ g Absent/ g Absent/ g Absent/ g Absent/ g
15 Salmonella API Absent/ g Absent/ g Absent/ g Absent/ g Absent/ g

1% of the Preservative Formulation 3 (comprising of 0.5% of AOC, 1% of AOE, AMPC2, AMPC3 each along with 0.5% of SSC1) is added to the Conditioner base making the total composition to 100% and this was tested for the 6 months accelerated stability accelerated stability. The color, odor, texture of the product was stable throughout the testing process. pH varied between 5.0 to 5.1 which is within the standard limits. There was no phase separation in the thermal stability testing. There was no microbial growth seen in the product by the end of the 6th month study period. Overall, there were no significant changes in the parameters tested at accelerated test duration of 6 months which, as per ICH Guidelines, can give a 24 Month i.e., 2-year shelf life to the product, thereby proving the potency of the Preservative Formulation-Table 12.

Example 7: Face Cream with the Preservative Formulation- Table 13
S. No. Test Parameter Method 0th Day 30th Day 60th Day 90th Day 180th Day
1 Appearance As per Standard Method Light pink
coloured cream Light pink
coloured cream Light pink
coloured cream Light pink
coloured cream Light pink
coloured cream
2 Odour As per Standard Method Characteristic Characteristic Characteristic Characteristic Characteristic
3 Texture As per Standard Method Soft Soft Soft Soft Soft
4 Colour As per Standard Method Light pink Light pink Light pink Light pink Light pink
5 pH IS 6608 4.924 5.045 5.076 5.070 5.087
6 Specific Gravity As per Standard Method 0.98 0.97 0.96 0.96 0.97
7 Centrifugation As per Standard Method No layer separation No layer separation No layer separation No layer separation No layer separation
8 Viscosity As per Standard Method 89800 cps 91240 cps 92000 cps 93600 cps 94800 cps
9 Total Yeast & Mould count lS 14648 <10 cfu/q <10 cfu/q <10 cfu/q <10 cfu/q <10 cfu/q
10 E.Coli lS 14648 Absent/ g Absent/ g Absent/ g Absent/ g Absent/ g
11 Salmonella ISO 6579 Absent/ g Absent/ g Absent/ g Absent/ g Absent/ g
12 Pseudomonas aeruginosa lS 14648 Absent/ g Absent/ g Absent/ g Absent/ g Absent/ g
13 Staphylococcus aureus lS 14648 Absent/ g Absent/ g Absent/ g Absent/ g Absent/ g

1% of the Preservative Formulation 3 (comprising of 0.5% of AOC, 1% of AOE, AMPC2, AMPC3 each along with 0.5% of SSC1) is added to the Face Cream base making the total composition to 100% and this was tested for the 6 months accelerated stability accelerated stability. The color, odor, texture of the product was stable throughout the testing process. pH varied between 4.9 to 5.0 which is within the standard limits. There was no phase separation observed in the Centrifugation process. There was no microbial growth seen in the product by the end of the 6th month study period. Overall, there were no significant changes in the parameters tested at accelerated test duration of 6 months which, as per ICH Guidelines, can give a 24 Month i.e., 2-year shelf life to the product, thereby proving the potency of the Preservative Formulation-Table 13.
Based on the above testing parameters and reports, it is evident that the product was stable throughout the test period and has passed the 6 months accelerated stability testing.
In one embodiment of the invention, higher ORAC Value depicted that the final preservative formulations had higher activity to combat the free radicals present in the substrate and thereby helped in improving the shelf life of the target product.

Table 14: Comparison of ORAC result between Individual and Combination of Phytocomponents

Phytocomponent ORAC
AOC 2502.4 mmol
AOE 61.52 mmol
AOL 13.26 mmol
AOE+AOC 4401.6 mmol
AOL+AOC 4377.5 mmol

Similarly, lower MIC values depicted that the ingredients in the final preservative formulation worked as effective antimicrobial agents, thereby increasing the shelf life of the target product.
In all Preservative Formulations (1-4), the microbial load was reduced and the final product was stable for longer periods when compared to control as seen in the result tables.
According to an embodiment of the invention, the final preservative formulation of phytocomponents helped in getting a longer shelf life when compared to the individual activity of the phytocomponents.
The results of the present invention have established that combining certain phytocomponents results in a synergistic antimicrobial effect, even at lower concentrations, compared to the use of individual substances.
The minimum inhibitory concentrations (MICs) of the preservative composition blend against various microbes were lower than those of the individual components
Table 15: Comparison of MIC result between Individual and Combination of Phytocomponents
Phyto component MIC
AMPC1 Gram Positive Bacteria - 15 mg/ml
Gram Negative Bacteria - 60 mg/ml
Fungi - 4 mg/ml
AMPC2 Gram Positive Bacteria - 16605 mg/ml
Gram Negative Bacteria - 62.4 mg/ml
Fungi - 12 mg/ml
AMPC3 Gram Positive Bacteria - 5.36 mg/ml
Gram Negative Bacteria - 5.36 mg/ml
Fungi - 32 mg/ml
AMPC1 + AMPC3 Gram Positive - 6.93 mg/ml
Gram Negative - 18.18 mg/ml
Fungi - 12.85 mg/ml
AMPC1 + AMPC2 Gram Positive - 12475 mg/ml
Gram Negative - 32 mg/ml
Fungi - 7.25 mg/ml
AMPC2 + AMPC3 Gram Positive - 12477.68 mg/ml
Gram Negative - 19.25 mg/ml
Fungi - 5.25 mg/ml

According to another embodiment of the invention, the extraction method followed to obtain the final preservative formulation helped in reducing the cost of the final product.
The compositions described herein may be formulated with other ingredients of the personal care product base such as water, emulsifiers, thickeners, surfactants, humectants, moisturizers, other botanicals, emollients, chelating agents and essential oils.
The product base contains botanicals from other herbs like Aloe barbadensis (Aloe Vera); Acacia concinna (Shikakai); Sapindus mukorossi (Reeta); Phyllanthus emblica (Amla); Rubia cordifolia (Manjista); Eclipta alba (Bhringaraj); Lawsonia inermis (Henna); Bacopa monnieri (Brahmi); Hibiscus Rosa sinensis (Hibiscus); Cedrus deodara (Devadaru); Trichosanthes dioica (Patola); Bacopa monnieri (Brahmi); Prunus amygdalus (Almond); Symplocos racemosa (Lodhra); Vetivera zizanoides (Usheera); Tephrosia purpurea (Sharpunkha); Curcuma aromatica (Turmeric); Orange peel; Azadirachta indica (Neem); Pterocarpus santalinus (SandalWood); Rosa centifolia (Rose); Crocus Sativus (Saffron); Withania somnifera (Ashwagandha); Cocos nucifera (Narikela); Elletaria cardamomum (Ela); Woodfordia floribunda (Dhataki); Syzygium cumini (Jambu); Salmalia malabarica (Mocharasa); Vateria indica (Sal Tree); Salix Alba (Vetasa); Pongamia glabra (Karanja) which are added to provide utmost efficacy of the product like Cleansing, Hair Growth, Skin Glow etc.
Foaming agents like Coco Glucoside; Lauryl Glucoside; Capryl/ Caprylyl Glucoside used for foaming and cleansing.
Moisturizing and Conditioning Agents like Shea Butter (Shea Tree); Olive oil; Glycerin; Xylitylglucoside (and) Anhydroxylitol (and) Xylitol; Almond Oil; Sunflower Oil; Jojoba Oil; Cocoa Butter; Sesame Oil; Decyl Oleate; Cetearyl Olivate (and) Sorbitan Olivate; Coco Caprylate; Glyceryl Stearate Citrate.
Chelating & Thickening Agents like Sodium Gluconate; Xanthan Gum.
Essential Oils like Jasmine Oil; Musk Oil; Ylang Ylang Oil; Rose Oil; Bergamot Oil; Tea Tree Oil; Orange Oil are used for enhancing the fragrance.

ADVANTAGES OF THE INVENTION
The present invention provides a natural preservative formulation for improving the shelf-stability of perishable personal care compositions. The additive effect of ‘synergistic’ actions of Phyto-components Eugenol, Curcumin, Linalool, Carnosic acid and moisture reducing natural salts Sodium Chloride and Sodium Bicarbonate shows enhanced anti-oxidant and anti-microbial activity and improved shelf-stability of perishable personal care products for 2 years.
The formulation is eco-friendly since the ingredients utilized are natural and free from synthetic chemicals.
The method of preparing the natural preservative formulation is simple as it involves a single extraction step by boiling the dried herbs in water and mixing the distillates in equal proportion to a salt in a fixed ratio to obtain the preservative extract and is less time consuming.
The preservative formulations of this patent application inhibit the growth of some of the microbes at very low concentration thereby giving a good shelf-life product of 2 years with no side-effects like skin irritation etc. The formulation is completely safe to be used on a day-to-day basis as the ingredients are added in appropriate percentages.
It is to be understood that the phraseology or terminology employed herein is for the purpose of description and not of limitation. Therefore, while the embodiments herein have been described in terms of preferred embodiments, those skilled in the art will recognize that the embodiments herein can be practiced with modification within the spirit and scope of the claims presented in the complete specification or non-provisional application.

,CLAIMS:We claim:
1. A polyherbal preservative formulation for the stabilization of topical personal care products comprising:
a) Selecting and processing a plurarity of herbs for their extracts; and wherein the said herbs are selected from the group comprising Curcuma spp., Rosmarinus spp., Syzygium spp., Ocimum spp., Zingiber spp., Lavandula spp. and Rosa spp.;
b) A distillate containing an aqueous extract from the rhizome of Curcuma spp. having the antimicrobial and antioxidant phytocomponent Curcumin;
c) A distillate containing an aqueous extract from the aerial plant parts of Rosmarinus spp. having the antimicrobial and antioxidant phytocomponent Carnosic Acid;
d) A distillate containing an ‘polyherbal aqueous extract’ obtained from the dried buds of Syzygium spp., the aerial plant parts of Ocimum spp. and the rhizome of Zingiber spp., having the antimicrobial and antioxidant phytocomponent Eugenol;
e) A distillate containing a ‘polyherbal aqueous extract’ obtained from the aerial plant parts of Ocimum spp., aqueous extract from floral parts of Rosa spp. and aqueous extract from the floral parts of Lavandula spp., containing the antimicrobial and antioxidant phytocomponent Linalool;
f) A salt selected from a group comprising Sodium chloride, Sodium Bicarbonate, etc.; and wherein the salt component is present as 0.5 % by weight; and
g) Water or oil;
and wherein the distillate of the aqueous extract from the rhizome of Curcuma spp. is in the range of 0.5-1% by weight, wherein the distillate of the aqueous extract from the aerial plant parts of Rosmarinus spp. is 1% by weight, wherein the polyherbal distillate obtained from the dried buds of Syzygium spp., the aerial plant parts of Ocimum spp. and the rhizome of Zingiber spp. are in the range of 0.5-2% by weight, and wherein the polyherbal distillate obtained from the aerial plant parts of Ocimum spp. , aqueous extract from the floral parts of Rosa spp., the floral parts of Lavandula spp. are present in the range between 0.5-1% by weight.
2. The naturally derived polyherbal preservative formulation as claimed in 1, wherein the distillates from the rhizome of Curcuma spp., wherein the distillate of the aqueous extract from the aerial plant parts of Rosmarinus spp., wherein the polyherbal distillate from the dried buds of Syzygium spp., the aerial plant parts of Ocimum spp. and the rhizome of Zingiber spp., and wherein the polyherbal distillate from the aerial plant parts of Ocimum spp., floral parts of Rosa spp., and the floral parts of Lavandula spp. are mixed in the ratio of 1:1:1:1.
3. The naturally derived polyherbal preservative formulation as claimed in 1, wherein the distillate from the rhizome of Curcuma spp., wherein the polyherbal distillate obtained from dried buds of Syzygium spp., the aerial plant parts of Ocimum spp. and the rhizome of Zingiber spp., wherein the polyherbal distillate from the aerial plant parts of Ocimum spp., floral parts of Rosa spp., and the floral parts of Lavandula spp., are mixed in the ratio of 0.5:2:1.
4. The naturally derived polyherbal preservative formulation as claimed in 1, wherein the distillate from the rhizome of Curcuma spp., wherein the distillate from the aerial plant parts of Rosmarinus spp.,wherein the polyherbal distillate obtained from the dried buds of Syzygium spp., the aerial plant parts of Ocimum spp. and the rhizome of Zingiber spp., wherein the polyherbal distillate obtained from the aerial plant parts of Ocimum spp., aqueous extract from floral parts of Rosa spp. and aqueous extract from the floral parts of Lavandula spp., are mixed in the ratio of 1:1:0.5:0.5.
5. The naturally derived polyherbal preservative formulation as claimed in 1, wherein the final preservative formulation is present up to 2% by weight of the total topical personal care product composition such as but not limited to shampoos, creams, washes, conditioners etc.

6. The naturally derived polyherbal preservative formulation as claimed in 1, wherein the pH level is in the range of 3-7.
7. A method of synthesizing a natural polyherbal preservative formulation for the stabilization of topical personal care products comprising the steps of:
a. Selecting and preparing a plurality of herbs for their extracts; and wherein the said herbs are selected from a group comprising Curcuma spp., Rosmarinus spp., Syzygium spp., Ocimum spp., Zingiber spp., Lavandula spp. and Rosa spp. [291];
b. Preparing a ‘distillate extract’ from the dried rhizomes of Curcuma spp. [292];
c. Preparing a ‘distillate extract’ from the dried aerial plant parts of the plant Rosmarinus spp. [292];
d. Preparing a ‘polyherbal distillate mixture’ from the dried buds of Syzygium spp., aerial plant parts of Ocimum spp., the rhizome of Zingiber spp. [292];
e. Preparing a ‘polyherbal distillate mixture’ from the aerial plant parts of Ocimum spp., the floral parts of Rosa spp. and the floral parts of Lavandula spp. [292];
f. Mixing the ‘distillate extract’ from the dried rhizomes of Curcuma spp.; ‘distillate extract’ from the dried aerial plant parts of the plant Rosmarinus spp.; ‘polyherbal distillate mixture’ from the dried buds of Syzygium spp., aerial plant parts of Ocimum spp., the rhizome of Zingiber spp.; and the ‘polyherbal distillate mixture’ from the aerial plant parts of Ocimum spp., the floral parts of Rosa spp. and the floral parts of Lavandula spp., in a particular proportion to obtain a homogenous mixture [293]; and
g. Mixing the ‘Homogenous Mixture’ so obtained with the salt selected from a group comprising Sodium chloride, Sodium Bicarbonate, etc. 0.5% by weight, and adjusting the pH level to fall in the range of 3-7 [294].
8. The method as claimed in 7, wherein the steps of selecting and preparing a plurality of herbs includes cleaning, drying, and subjecting the dried herbs to sublimation of moisture through freeze-drying and storage under liquid nitrogen, wherein the drying of the herbs is carried out for 2-3 days in shade [291].
9. The method as claimed in 7, wherein the distillates are obtained by boiling 10g of the said plurality of herbs in 60 ml of water at 80? with a vacuum pressure of 600mm of Hg and the distillates are collected separately [292]; and wherein the distillates are mixed in the ratio of 0.5:1:1:0.5, or in the ratio of 1:1:1:1, or in the ratio of 0.5: 0:2:1 [293].
10. The method as claimed in 7, wherein the ratio of the herbs used to make the poly herbal distillates for the distillation process is in equal proportion, by weight [292].