DESC:FIELD OF INVENTION

[001] The present invention relates to a herbal tea composition and a method for preparation thereof. Particularly, the present invention relates to a herbal tea composition exhibiting antibacterial, antiviral and anti-inflammatory properties which relieves throat related infections and associated symptoms thereof.

BACKGROUND OF THE INVENTION
[002] With an increase in occurrence of infections on a day-to-day basis, it has become essential for people to strengthen their immune system, resulting in efficient functioning of
the respiratory tract. Such infections are caused by harmful microorganisms entering the human body. Such disease causing harmful microorganisms are known as pathogens. Some major pathogens causing infections in humans include bacteria and viruses.

[003] Herbs exhibit a potential to enhance overall health as it possess antimicrobial and anti-inflammatory properties that helps in fighting off the infections, thereby enhancing the immune response of the body. The secondary metabolites present in the herbs exhibit a desired efficacy in improving immune response of the body. The herbs containing such metabolites are consumed in different forms such as, but not limited to, extracts, powders, capsules, liquid and whole plants. One such liquid form includes tea containing a combination of different herbs that contain a plurality of secondary metabolites. Such herbal teas enhances the immune response of the body.

[004] Herbal teas are a type of beverages prepared from infusion or decoction of herbs, spices and/or other plant material in hot water. Such decoction helps in fighting off various harmful infections. The herbal teas also provide relief to sore throat and acts as an expectorant and mucolytic.

[005] There are several patent applications providing a herbal tea composition that helps in preventing various throat related infections and other diseases. A Chinese Patent Application No. CN1098876A entitled Health-care licorice root tea, wherein it discloses the health-care liquorice tea is made up of liquorice root, fleece flower root, chrysanthemum flower and tea through slicing clean liquorice root and fleece flower root, baking, breaking, sieving, and mixing with chrysanthemum flower and tea, and package. The disclosed herbal tea composition provides the benefits to the heart, lung, brain and anti-ageing effect. However, the disclosed herbal tea may not effective in treating sore throat and other throat related infections as the herbal tea disclosed herein do not incorporate biologically active compounds present in different parts of the herbs possessing anti-inflammatory, anti-bacterial and other therapeutic properties that aids in treating sore throat and related infections. Further, the disclosed herbal tea includes only limited number of plants based ingredients. It fails to include other herbal extracts such as Withania somnifera (Ashwagandha), Panax ginseng (Asian ginseng), Ocimum basilicum (Basil), Ginkgo biloba (Ginkgo), and Zingiber officinale (Ginger). Moreover, the disclosed herbal tea do not incorporate plant based natural flavoring agent conferring a desirable flavor by altering taste and smell of the items.

[006] In view of the above problems associated with the state of the art, there remains a need to provide an effective herbal tea composition that relieves sore throat and associated symptoms thereof.
OBJECTIVES OF THE INVENTION
[007] The primary objective of the present invention is to provide a herbal tea composition and a method for preparation thereof.

[008] Another objective of the present invention is to enhance immune response of the body by providing a herbal tea composition including a number of herbs that exhibits medicinal properties.

[009] Yet another objective of the present invention is to provide a herbal tea composition exhibiting antibacterial, antiviral and anti-inflammatory properties which relieves sore throat and associated symptoms thereof.

[0010] Other objects and advantages of the present invention will become apparent from the following description taken in connection with the accompanying drawings, wherein, by way of illustration and example, the aspects of the present invention are disclosed.

SUMMARY OF THE INVENTION

[0011] The present invention relates to a herbal tea composition and a method for preparation thereof. The herbal tea composition of the present invention comprises of one or more herbal extracts, one or more flavoring agents, an active ingredient, a bioenhancer, and a solvent.. The herbal extracts are present in a range of 0.2% to 90% by weight of the total herbal tea composition. The flavoring agents are present in a range of 0.1% to 5% by weight of the total herbal tea composition. The active ingredient i.e. the hydroalcoholic extract containing parts of the herbs loaded with the active ingredients are present in a range of 1% to 80% by weight of the total herbal tea composition. . The bioenhancers are present in a range of 0.001% to 1% by weight of the total herbal tea composition. The solvents are present in a range of 1% to 10% by weight of the total composition. The method for preparing the herbal tea composition is based on granulation technique employing natural bioenhancers and solvent, resulting in the formation of granules from of a mixture consisting of different herbal extracts. The granulation technique enhances the absorption and dissolution of the secondary metabolites present in the composition.. The herbal tea composition exhibits antibacterial, antiviral and anti-inflammatory properties which relieves throat related infections and associated symptoms thereof.
SOURCE OF BIOLOGICAL MATERIAL USED IN THE INVENTION
[0012] The herbal extracts used in the present invention including such as, but not limited to, Withania somnifera, Ocimum basilicum, Panax ginseng, Ginkgo biloba, Glycyrrhiza glabra, and Zingiber officinale are procure from local vendors. The flavoring agents used in the present invention such as, but not limited to, Basil flavor, Ashwagandha flavor, Orange flavor, Cinnamon flavor, Cardamom flavor, Anise flavor, Lemon flavor, Ginger flavor, Cardamom flavor and clove flavor are also procured from local vendors. The parts of herbs for preparing the hydroalcoholic extract used in the present invention such as, but not limited to, fruit of Black Pepper (Piper nigrum), bark of Cinnamon (Cinnamomum zeylanicum), bud of Clove (Eugenia caryophyllus), bud of Cardamom (Elettaria cardamomum), leaf of tea plant (Camellia sinensis) and flower of Pomegranate (Punica granatum) are also procured from the vendors. The bioenhancers are derived from different parts of herbs such as, but not limited to, Ginger (Zingiber officinale) and Basil (Ocimum basilicum) are procured from local vendors. The test strains used for determining microbial contamination including Salmonella abony was procured from National Collection of Type Cultures with NCTC number – NCTC6017 and Pseudomonas aeruginosa is procured from American Type Culture Collection (ATCC) with ATCC number – ATCC9027. The Wistar rat for evaluating acute inhalation toxicity of the composition was procured from Genotox Bio Services Pvt. Ltd (CPCSEA registration No.: 1242/PO/RcBiBt/S/08/CPCSEA).

BRIEF DESCRIPTION OF DRAWINGS
[0013] An understanding of the present invention may be obtained by reference to the accompanying drawings, when taken in conjunction with the description herein and in which:

[0014] Figure 1 illustrates graphical representation depicting germ kill efficacy of the herbal composition against common pathogens;

[0015] Figure 2 (a) and Figure (b) illustrates graphical representation depicting anti-inflammatory activity of the herbal composition;

[0016] Figure 3 illustrates graphic representation depicting analgesic potential of the herbal composition;

[0017] Figure 4 (a) and Figure 4(b) illustrates graphic representation depicting mucolytic potential of the herbal composition; and

[0018] Figure 5 illustrates graphic representation depicting antioxidant potential of the herbal composition.
DETAILED DESCRIPTION OF THE INVENTION
[0019] The following description describes various features and functions of the disclosed invention. The illustrative embodiments described herein are not meant to be limiting. It may be readily understood that certain embodiments of the disclosed system can be arranged and combined in a wide variety of different configurations, all of which have not been contemplated herein.

[0020] Accordingly, those of ordinary skill in the art will recognize that various changes and modifications of the embodiments described herein can be made without departing from the scope of invention. In addition, descriptions of well-known functions and constructions are omitted for clarity and conciseness.

[0021] Features that are described and/or illustrated with respect to one embodiment may be used in the same way or in a similar way in one or more other embodiments and/or in combination with or instead of the features of the other embodiments.

[0022] The terms and words used in the following description are not limited to the bibliographical meanings, but, are merely used to enable a clear and consistent understanding of the invention. Accordingly, it should be apparent to those skilled in the art that the following description of exemplary embodiments of the present invention are provided for illustrative purpose only and not for the purpose of limiting the invention. The term "herbal composition" is used interchangeably with the term "herb composition" and refers to any composition derived from a plant source.

[0023] It is to be understood that the singular forms “a”, “an” and “the” include plural referents unless the context clearly dictates otherwise.

[0024] It should be emphasized that the term “comprises/comprising” when used in this specification is taken to specify the presence of stated features, steps or components but does not preclude the presence or addition of one or more other features, steps, components or groups thereof.

[0025] Accordingly, the present invention relates to a herbal tea composition and a method for preparation thereof. Particularly, the present invention relates to a herbal tea composition exhibiting antibacterial, antiviral and anti-inflammatory properties which relieves throat related infections and associated symptoms thereof.

[0026] In an embodiment, the herbal tea composition of the present invention comprises of one or more herbal extracts, one or more flavoring agents, an active ingredient, a bioenhancer, and a solvent.

[0027] The herbal extract is a solution containing a plurality of herbs dissolved in a solvent. The herbal extract is prepared by separating mixture containing a number of secondary metabolites present in the herbs using a solvent. Such extraction process results in separation of soluble secondary metabolites present in the herbs and insoluble cellular portion. The secondary metabolites are bio-active chemical substances produced by plants. Such metabolites may be classified into various categories such as, but not limited to, terpenoids, alkaloids, phenolic compounds and so on. The secondary metabolites present in the herbal extracts possess antibacterial, antiviral and anti- inflammatory properties, thereby imparting medicinal properties to the herbal extracts. In an exemplary embodiment, the herbal extracts used in present invention includes the extracts of herbs selected from a group consisting of, such as, but not limited to, Withania somnifera (Ashwagandha), Ocimum basilicum (Basil), Panax ginseng (Asian ginseng), Ginkgo biloba (Ginkgo), Glycyrrhiza glabra (Licorice) and Zingiber officinale (Ginger) or a combination thereof. The herbal extracts used in the present invention exhibits antibacterial, antiviral and anti-inflammatory properties imparted by the secondary metabolites. In an exemplary embodiment, the secondary metabolites present in the herbal extracts may include such as, but not limited to, withanolide present in Withania somnifera, terpenes and flavonoids present in Ocimum basilicum, Ginsenoside present in Panax ginseng, flavones present in Ginkgo biloba and Gingerol present in Zingiber officinale and so on. In an embodiment, a pre-prepared alcoholic or hydroalcoholic herbal extract may be used in the present invention. The herbal extracts are present in a range of 0.2% to 90% by weight of the total herbal tea composition. The secondary metabolites in the herbal extracts are present in a range of 1% to 25% by weight of the total herbal extracts. The herbal extracts used in the present invention are described in detail herein:

[0028] (a) Ashwagandha Extract - Ashwagadha (Withania somnifera) possesses immunity enhancing properties, thus strengthening the immune response of the body. Ashwagadha also reduces weakness and fatigue. The antioxidant and analgesic properties of Ashwagadha aids in treating sore throat, cold, cough, etc. Ashwagadha enhances the energy and body strength, thereby helping the patients to recover from severe illness.

[0029] (b) Basil Extract - Basil (Ocimum basilicum) exhibits antimicrobial, anti- inflammatory, antibiotic and antioxidant properties that provides relief from throat related infections and associated symptoms thereof. The antibacterial and antiviral property possessed by Basil provides protection against harmful pathogen causing throat related infections. The anti-inflammatory property of Basil aids in treating swelling of the throat tissues. Basil also enhances the immune response of the body.

[0030] (c) Ginseng Extract – Ginseng (Panax ginseng) exhibits antiviral effects and thus inhibits viral attachment, membrane penetration, and replication. Ginseng also possesses antioxidant and anti-inflammatory properties. The anti-inflammatory property of Ginseng aids in reducing swelling of the throat tissues. Ginseng also enhances the immune response of the body.

[0031] (d) Ginkgo Extract – Ginkgo (Ginkgo Biloba) contains potent antioxidants that aids in fighting off deleterious effects of free radicals. Ginkgo also aids in improving asthma. The anti-inflammatory property of Ginkgo provides relief from sore throat, bronchitis and various other inflammatory respiratory diseases such as chronic obstructive pulmonary disease (COPD). The antioxidant property of Ginkgo helps in reducing inflammation of airways. Ginkgo also increases the flow of blood to various parts of the body.

[0032] (e) Licorice extract – Licorice (Glycyrrhiza glabra) exhibits antimicrobial, antioxidant and anti-inflammatory properties, thereby providing relief from throat related infections and associated symptoms thereof. Licorice aids in treating sore throat, hoarseness of voice, cough and other respiratory conditions. Licorice also aids in reducing inflammation in the airways. It also loosen mucus in lungs, thereby clearing congestion.

[0033] (f) Ginger extract- Ginger (Zingiber officinale) exhibits anti- inflammatory, antiviral, antibacterial, antioxidant properties. Such properties help in curing cold, congestion, cough and flu. Gingers also provides relief from sore throat and other throat related infections. The anti-inflammatory property of Ginger aids in reducing inflammation.

[0034] The flavoring agents are the compounds added to a beverage or food items to confer a desirable flavor by altering taste and smell of the items. In a preferred embodiment, the flavoring agents used in the present invention may include plant based natural flavoring agents, which are derived from different herbs. In an exemplary embodiment, the flavoring agents may be selected from a group consisting of, such as, but not limited to, Basil flavor, Ashwagandha flavor, Orange flavor, Cinnamon flavor, Cardamom flavor, Anise flavor, Lemon flavor, Ginger flavor, Cardamom flavor, and Clove flavor, or a combination thereof. The flavoring agents are present in a range of 0.1% to 5% by weight of the total herbal tea composition. In an exemplary embodiment, the flavoring agents are present in different forms such as but not limited to, granules, power, and liquid.

[0035] The active ingredient used in the present invention is biologically active compound present in different parts of different herbs. Such compound possesses antibacterial, antiviral and anti-inflammatory properties which relieves throat related infection and associated symptoms thereof. In a preferred embodiment, the active ingredient used in the present invention is in form of a ready-made hydroalcoholic extract containing parts of the herbs loaded with active ingredients. In an exemplary embodiment, the parts of the herbs containing active ingredients used for preparing the hydroalcoholic extract may be selected from a group consisting of, such as, but not limited to, fruit of Black Pepper (Piper nigrum), bark of Cinnamon (Cinnamomum zeylanicum), bud of Clove (Eugenia caryophyllus), bud of Cardamom (Elettaria cardamomum), leaf of tea plant (Camellia sinensis) and flower of Pomegranate (Punica granatum), or a combination thereof. In another exemplary embodiment, the active ingredient present each part of each herb may include such as, but not limited to, Piperine present in Piperum nigrum, Cinnamonaldehyde in Cinnamomum zeylanicum, Eugenol in Eugenia caryophyllus, Polyphenols in Elettaria cardamomum, Tannins and flavonoids in Camellia sinensis; and Punicalagin in Punica granatum The active ingredient i.e. the hydroalcoholic extract containing parts of the herbs loaded with the active ingredients are present in a range of 1% to 80% by weight of the total herbal tea composition. In another exemplary embodiment, total active ingredients present in different parts of the herbs are in range of 0.25 % to 90% by weight of the all parts of the herbs. The properties of the herbs containing the active ingredient are described in detail herein:

[0036] (a) Black Pepper – The active ingredient, Piperine, in black pepper (Piper nigrum) exhibits anti-inflammatory and antioxidant properties. Such properties help in curing respiratory disorders such as cough, cold, asthma, stuffy nose, bronchitis sinus infection. It also helps in reducing inflammation and improves breathing.

[0037] (b) Cinnamon - Cinnamon (Cinnamomum zeylanicum) possesses anti-inflammatory property imparted by Cinnamonaldehyde, which aids in reducing inflammation. Cinnamon also exhibits antimicrobial property that aids suppressing/inhibiting growth of various microorganisms such as virus, bacteria and fungi. Cinnamon also strengthens the immune response of body and aids in fighting off free radicals, thereby protecting against germs causing respiratory disorders.

[0038] (c) Clove – Clove (Eugenia caryophyllus) possesses antimicrobial properties imparted by Eugenol, which aids in inhibiting the growth of microorganisms. The Clove also exhibits anti-inflammatory properties which aids in curing cold, cough, sore throat and sinusitis.

[0039] (d) Cardamom – Cardamom (Elettaria cardamomum) possesses antioxidants imparted by Polyphenols, which aids in improving overall health. Cardamom exhibits antimicrobial property that aids in preventing the growth of microorganisms. Cardamom also possesses antiseptic and anti-inflammatory properties that aids inreducing swelling and pain in the membrane of mucus, throat and mouth.

[0040] (e) Tea leaves – The leaves of tea plant (Camellia sinensis) possess a number of antioxidants imparted by tannins and flavonoids. It aids in fighting off free radicals, thereby protecting against germs causing respiratory disorders.

[0041] (f) Pomegranate - Pomegranate (Punica granatum) possesses a number of antioxidants that aids in removing free radicals and safegaurd cells from damage. Pomegranate also possesses anti-inflammatory and anti-microbial, properties that aids in reducing inflammation and prevents the growth of microorganisms.

[0042] The bioenhancers enhance the bioavailability of the extracts in the human body, thereby increasing bio-efficacy thereof. In a preferred embodiment, natural bioenhancers are used in the present invention. In an exemplary embodiment, the natural bioenhancers used in the present invention are present in different parts of the herb such as, but not limited to, Zingiber officinale (Ginger) and Ocimum basilicum (Basil). In another preferred embodiment, a combination of Zingiber officinale (Ginger) and Ocimum basilicum (Basil) in powered form is used as bioenhancer in the present invention. The bioenhancers are present in a range of 0.001% to 1% by weight of the total herbal tea composition.
[0043] The solvent used in the present invention aids in granulating the herbal extracts used in the present invention. In an exemplary embodiment, the solvent may be selected from a group consisting of, such as, but not limited to, Isopropyl alcohol (IPA), or ethyl alcohol. The solvents are present in a range of 1% to 10% by weight of the total herbal tea composition.

[0044] In an exemplary embodiment, Table 1 illustrates different compounds of the composition and secondary metabolites present in several compounds along with the amount of each compound (ml).

Sr.
No. Components Quantity (ml) Secondary metabolites/ Active Ingredient
1. Basil extract 0.0 - 0.75 Terpenes and flavonoids
2. Piper nigrum 0.0 - 0.25 Piperine
3. Ashwagandha extract 0.0 - 0.75 Withanolide
4. Cinnamon bark 0.0 - 0.25 Cinnamonaldehyde
5. Ginseng extract 0.0 - 0.75 Ginsenoside
6. Anise seed extract 0.0 - 1.25
7. Liquorice extract 0.0 - 0.75 Glycyrrhizin
8. Clove bud 0.0 - 0.25 Eugenol
9. Cardamom bud 0.0 - 0.25 Polyphenols
10. Ginger extract 0.0 - 0.5 Gingerol
11. Anar flower 0.0 - 2.0 -
12. Basil GF 0.0 - 1.75 -
13. Ashwagandha GF 0.0 - 1.75 -
14. Orange GF 0.0 - 1.25 -
15. Cinnamon GF 0.0 - 1.25 -
16. Clove GF 0.0 - 1.25 -
17. Cardamom GF 0.0 - 1.25 -
18. Anise EC 0.0 - 0.40 -
19. Lemon EC 0.0 - 1.25 -
20. Ginger GF 0.0 - 1.25 -
21. Cardamom OS 0.0 - 1.50 -
22. Tea leaves 15.0 - 79.35 Tannins and flavonoids
Table 1
[0045] In another embodiment, the present invention also provides a method for preparation of a herbal tea composition. The method is based on granulation technique employing natural bioenhancers and solvent, resulting in the formation of granules from of a mixture consisting of different herbal extracts. The granulation technique enhances the absorption and dissolution of the secondary metabolites present in the composition. The method is described herein detail:

a) adding herbal extracts in an apparatus;
b) straining the herbal extracts through a sieve;
c) granulating the extracts obtained in step (b) with a solvent and bioenhancer, resulting in the formation of granules;
d) allowing the granules to dry at a temperature in a range of 20? to 50? for 3 to 4 hours;
e) adding flavoring agents and the active ingredient to the granules obtained in step (d); and
f) mixing the mixture obtained (d) thoroughly, forming a herbal composition.;

[0046] The time period for drying the granules formed after granulation technique may depend on the quantity and moisture value of the granules.

[0047] In an exemplary embodiment, the herbal extracts used in present invention includes the extracts of herbs selected from a group consisting of, such as, but not limited to, Withania somnifera (Ashwagandha), Ocimum basilicum (Basil), Panax ginseng (Asian ginseng), Ginkgo biloba (Ginkgo), Glycyrrhiza glabra (Licorice) and Zingiber officinale (Ginger) or a combination thereof. . In an embodiment, a pre-prepared alcoholic or hydroalcoholic herbal extract may be used in the present invention. The herbal extracts are present in a range of 0.2% to 90% by weight of the total herbal tea composition.

[0048] In a preferred embodiment, the flavoring agents used in the present invention may include plant based natural flavoring agents, which are derived from different herbs. In an exemplary embodiment, the flavoring agents may be selected from a group consisting of, such as, but not limited to, Basil flavor, Ashwagandha flavor, Orange flavor, Cinnamon flavor, Cardamom flavor, Anise flavor, Lemon flavor, Ginger flavor, Cardamom flavor, and Clove flavor, or a combination thereof. The flavoring agents are present in a range of 0.1% to 5% by weight of the total herbal tea composition. In an exemplary embodiment, the flavoring agents are present in different forms such as but not limited to, granules, power, and liquid.

[0049] In a preferred embodiment, the active ingredient used in the present invention are in form of a ready-made hydroalcoholic extract containing parts of the herbs loaded with active ingredients. The active ingredient i.e. the hydroalcoholic extract containing parts of the herbs loaded with the active. In an exemplary embodiment, the parts of the herbs used for preparing the hydroalcoholic extract may be selected from a group consisting of, such as, but not limited to, fruit of Black Pepper (Piper nigrum), bark of Cinnamon (Cinnamomum zeylanicum), bud of Clove (Eugenia caryophyllus), bud of Cardamom (Elettaria cardamomum), leaf of tea plant (Camellia sinensis) and flower of Pomegranate (Punica granatum), or a combination thereof.

[0050] In a preferred embodiment, natural bioenhancers are used in the present invention. In an exemplary embodiment, the natural bioenhancers used in the present invention are present in different parts of the herb such as, but not limited to, Zingiber officinale (Ginger) and Ocimum basilicum (Basil). ). In another preferred embodiment, a combination of Zingiber officinale (Ginger) and Ocimum basilicum (Basil) in powered form are used as bioenhancers in the present invention. The bioenhancers are present in a range of 0.001% to 1% by weight of the total herbal tea composition

[0051] In an exemplary embodiment, the solvent may be selected from a group consisting of, such as, but not limited to, Isopropyl alcohol (IPA), or ethyl alcohol. The solvents are present in a range of 1% to 10% by weight of the total herbal tea composition.

[0052] In an exemplary embodiment, the herbal tea composition prepared in the present invention may be used in different forms such as, but not limited to, capsules, tablet, granules, powder and decoction. In a preferred embodiment, the herbal tea composition prepared in the present invention is used in a powdered form.

[0053] The following illustrates experimental data of the present invention and should not be construed to limit the scope of the invention.

Experimental Data:

[0054] The general parameters of the herbal tea composition of the present invention were evaluated such as color, odor, density, energy, moisture and sugar content. The herbal tea composition was perceived to be brown in color. The herbal tea composition exhibited a characteristic odor. The density of the herbal tea composition was evaluated to be 362.17 kcal/100g. The energy value of the herbal tea composition was evaluated to be 362.17 kcal/100g. The moisture content of the herbal tea composition was evaluated [as per IS 11623:2008 (RA2018)] to be 7.33% by weight of the total composition. The sugar content was not found in the herbal tea composition.

[0055] Table 2 illustrates experimental results of the tests performed for the evaluation of general parameters along with the standard limit.

Sr No. Test Result Standard Limits
1. Color Brown Brown
2. Odour Characteristic Characteristic
3. Density 0.405 gm/ml 0.2-0.5 gm/ml
4. Energy 362.17 kcal/100g -
5. Moisture 7.33%w/w -
6. Total Sugar Nil -
Table 2
[0056] The nutritional value of the herbal tea composition was evaluated. Some of the essential nutritional parameters such as amount of energy [as per IS 14433 :2007 (RA 2018)], fat [IS 14684 :1975 (RA 2020)], carbohydrate [IS 1656 :2007 (RA 2018)], protein [IS 7219 :1973 (RA 2015)] and sodium were evaluated. The content of the energy in the herbal tea composition was evaluated to be 362.17 kcal/100g. Similarly, the content of carbohydrates was evaluated to be 79.63 g/100g, protein was 7.34 g/100g and fat was 1.59 g/100g

[0057] Table 3 illustrates results of experimental results of the tests performed for the evaluation of the nutritional value of the herbal tea composition along with the standard limit.

Sr No. Test Result Standard Limits
1. Energy 362.17 kcal/100g -
2. Carbohydrate 79.63 g/100g -
3. Protein 7.34 g/100g -
4. Fat 1.59 g/100g -
Table 3
[0058] The amount of ash present in the herbal tea composition was evaluated [as per IS 4684 :1975 (RA 2020)]. The method for evaluating the amount of the ash present in the herbal tea composition involves adding 2.0 g of the sample of herbal tea composition into a previously dried crucible. The crucible was then kept in a muffle furnace at 100°C for 30 minutes. The temperature was then increased in 50°C increments up to 250°C at the intervals of 30 min. After 30 min, the temperature was further increased to 500 °C and the material was incinerated until the material turns white, indicating the absence of carbon. The crucible was cooled completely in a desiccator. The total amount of ash was weighed followed by calculating the percentage of total ash in reference to powder sample taken initially. The amount of ash in the herbal tea composition was evaluated to be 4.11% by weight of the total composition, which is within the standard limit of 3-5% by weight of the total composition.

[0059] The heavy metal analysis of the herbal tea composition was performed. The amount of a number of heavy metals such as Lead, Cadmium, Arsenic and Mercury was evaluated. The method for analysis of heavy metal was based on Atomic Absorption Spectrophotometry. The method involves transferring 3 g of the sample of the herbal tea composition into a clean, dry Kjeldahl flask of volume 300 ml. The flask was clamped at an angle of 45° followed by adding a desired amount of a concentrated nitric acid to moisten the substance thoroughly. The solution was then warmed until the reaction commenced, allowing the reaction to subside. The portions of same acid mixture was added to the prepared solution with constant heating after adding the acid each time until a total of 18 ml of the acid was added. The temperature was increased until the solution turned dark in color. The solution was then allowed to cool. 2 ml of nitric acid was added and heated again until the solution turned dark in color. The solution was constantly heated followed by the addition of nitric acid until no further darkening in the color of the solution occurred. The content of the solution was strongly heated to form dense, white fumes. 5 ml of water was added to cool the content of the solution. The solution was boiled gently to form dense, white fumes. The solution was then constantly heated until the volume of the solution was reduced to a few ml. 5 ml of water was added cautiously to cool the solution and the color of the solution was examined. In case, the solution was yellow in color, 1 ml of 30% hydrogen peroxide was added to the solution and again evaporated to form dense, white fumes and a volume of 2 to 3 ml. However, if the solution thus obtained was again yellow in color, 5 ml of water was added and the peroxide treatment was repeated. The content of the solution was then cooled, diluted cautiously with a few ml of water and rinsed into a 50 ml color- comparison tube, ensuring that the combined volume not exceeded 25 ml. A blank solution was prepared following the same procedure without utilizing the sample. At least, three standard solutions of different concentrations of the element being examined were prepared, covering 25 to 200% of the range that may be present in the sample solution. The blank reference solution was prepared with a separate set of corresponding reagents. The instrument was calibrated, operated as per manufacturer’s recommendations and analytical condition suitable for the analysis of lead, cadmium, arsenic, and mercury was set. The absorbance of the blank reference solution and each reference solution of different concentrations were separately measured and the readings were recorded. A calibration curve was prepared with an average value of three readings of each concentration on the ordinate and the corresponding concentration on the abscissa. The mean value of the readings obtained was interpolated with the test solution on the calibration curve to determine the concentration of each heavy metal. The amount of mercury, cadmium, lead and arsenic was reported to be below the quantification limit.

a. Table 4 illustrates the amount of heavy metals present in the herbal tea composition along with the standard limits.

Sr No. Test Result Standard Limits
1. Mercury Below Quantification
Limit 0.20-1.0ppm
2. Cadmium Below Quantification
Limit 0.20-0.30ppm
3. Lead Below Quantification
Limit 0.20-10.0ppm
4. Arsenic Below
Quantification Limit 0.20-3.0ppm
Table 4
[0060] The microbial count in the herbal tea composition was detected by evaluating microbial contamination of the herbal tea composition caused by yeast, mould, Escherichia coli, Pseudomonas aeruginosa, Salmonella abony and Staphylococcus aureus (as per API, Part-I, Vol.-IX (Extracts); Apeendix-3,3.2). The method is based on evaluating total viable count and detection of particular microbes. The method for detection of total viable count involves enumeration of colonies on a non-selective agar medium (Plate count). The count depict the number of colony forming units (cfu) per gram/ml of the sample. The probability of inhibition of growth of the microbes by the sample was then neutralized, thereby allowing detection of viable microorganisms. The method for detection of specific microorganisms such as Escherichia coli, Pseudomonas aeruginosa, Salmonella abony and Staphylococcus aureus involves performing enrichment by using a non-selective broth medium to increase the number of micro-organisms without any risk of inhibition by the selective ingredients present in selective/differential growth media. The isolation step was performed on a selective medium followed by identification tests. The total microbial count (cfu/g) was evaluated to be 85 cfu/gm and total yeast and mould (count/g) was evaluated to be 20 cfu/gm, both of which was within the standard limit. Further, Escherichia coli, Pseudomonas aeruginosa, Salmonella abony and Staphylococcus aureus were reported to be absent in the sample.

[0061] Table 5 illustrates the microbial contamination in the herbal tea composition along with the standard limits.

Sr No. Test Result Standard Limits
1. Total Microbial Count,
cfu/g 85 cfu/gm Not more than
100000
2. Total Yeast and Mould, count/g 20 cfu/gm Not more than 1000
3. Escherichia coli /g Absent Should be
absent
4. Pseudomonas
aeruginosa /g Absent Should be
absent
5. Salmonella abony/10g Absent Should be
absent
6. Staphylococcus
aureus/g Absent Should be
absent
Table 5
[0062] The amount of different types of aflatoxins such as, Aflatoxin B1, Aflatoxin B2, Aflatoxin G1 and Aflatoxin G2 present in the herbal tea composition was evaluated (as per API, Part-I, Vol.-IX (Extracts); Appendix-3,3.4). The method involves preparing 2.5 µl, 5 µl, 7.5 µl and 10 µl of the aflatoxin solution and three 10 µl applications of either test solution 1 or test solution 2 and applying to a suitable thin-layer chromatographic plate coated with a 0.25-mm layer of chromatographic silica gel mixture. The method further employs superimposing 5 µl of the aflatoxin solution on one of the three 10 µl applications of the test solution. The spots were allowed to dry. A chromatogram was developed in an unsaturated chamber containing a solvent system consisting of a mixture of chloroform, acetone and isopropyl alcohol in a ratio 85:10:5 until the solvent front moved at least 15 cm from the origin. The plate was removed from the developing chamber and the solvent front was marked and allowed to air dry. The spots on the plate were detected by examination under UV light at 365 nm. The four applications of the aflatoxin solution appeared as four clearly separated blue fluorescent spots. The spot obtained from the test solution superimposed on the aflatoxin solution was not as intense as compared to the corresponding aflatoxin solution. Further, the spots obtained from any of the other test solutions was not in corresponding to any of the spots obtained from the applications of the aflatoxin solution. In case, any spot of aflatoxins was obtained in the test solution then position of each fluorescent spot of the test solution was compared with the spots of the aflatoxin solution, thereby detecting the type of aflatoxin present in the herbal tea composition. In case, the aflatoxin spot was present in the test solution, the intensity of the aflatoxin spot as compared to the corresponding aflatoxin in the aflatoxin solution provide an approximate concentration of the aflatoxin in the test solution. The amount of total aflatoxin including Aflatoxin B1, Aflatoxin B2, Aflatoxin G2 and Aflatoxin G1 was reported to be below the quantification limit.

[0063] Table 6 illustrates the amount of the aflatoxin present in the herbal tea composition along with the standard limits.

Sr No. Test Result Standard Limits
1. Total Aflatoxin Below Quantification
Limit Not more than 5mcg/Kg
2. Aflatoxin B1 Below Quantification
Limit 1-2mcg/Kg
3. Aflatoxin B2 Below Quantification
Limit 1-2mcg/Kg
4. Aflatoxin G2 Below Quantification
Limit 1-2mcg/Kg
5. Aflatoxin G1 Below Quantification
Limit 1-2mcg/Kg
Table 6
[0064] The pesticidal residues present in the herbal tea composition were evaluated (as per API, Part-I, Vol.-IX (Extracts); Appendex-3,3.3). The method for evaluating the pesticidal residue employed gas chromatography. The gas flow rates specified in the monograph was maintained until a stable baseline was achieved. The test solution/s and the reference solution/s were prepared. The solutions did not contain solid particles. The samples were then run, and the quantitative results were evaluated. The amount of Alchalor, Malathoin, Phoslane, Pyrethrins, Ethion, Chlorpyrifos, Methyl Parathion, Lindane (Gamma - HCH), Aldrin and dieldrin, Azinophos – methyl, Bromopropylate, Chlorpyrifos-methyl, Deltamethrin, Diazinon, Dichlorvos, Endrin, Fonofos, Methidathion, Parathion, Permethrin , Perimiphos-methyl, Fentrothion, Cypermethrin and isomers, Hexachlorocyclohexane, Piperonyl-butoxide, Heptachlor, Fenvalerate, Endosulfan, DDT, Hexachlorobenzene, Chlorfenvinphos and Dithiocarbamates was reported to be below the quantification level.

[0065] Table 7 illustrates the amount of the pesticidal residue present in the herbal tea composition along with the standard limits.

Sr No. Test Result Standard Limits
1. Alchalor Below Quantification
Limit 0.0050-
0.020mg/kg
2. Malathoin Below Quantification
Limit 0.0050-
1.0mg/kg
3. Phoslane Below
Quantification Limit 0.0050-
0.10mg/kg
4. Pyrethrins Below
Quantification Limit 0.0050-
3.0mg/kg
5. Ethion Below Quantification
Limit 0.0050-2mg/kg
6. Chlorpyrifos Below Quantification
Limit 0.0050-
0.20mg/kg
7. Methyl Parathion Below Quantification
Limit 0.0050-
0.20mg/kg
8. Lindane (Gamma - HCH) Below
Quantification Limit 0.0050-
0.60mg/kg
9. Aldrin and dieldrin (sum of) Below
Quantification Limit 0.0050-
0.50mg/kg
10. Azinophos - methyl Below
Quantification Limit 0.0050-
1.0mg/kg
11. Bromopropylate Below
Quantification Limit 0.0050-
3.0mg/kg
12. Chlorpyrifos-methyl Below
Quantification Limit 0.0050-
0.10mg/kg
13. Deltamethrin Below
Quantification Limit 0.0050-
0.50mg/kg
14. Diazinon Below
Quantification Limit 0.0050-
0.10mg/kg
15. Dichlorvos Below Quantification
Limit 0.0050-
1.0mg/kg
16. Endrin Below Quantification
Limit 0.0050-
0.050mg/kg
17. Fonofos Below
Quantification Limit 0.0050-
0.050mg/kg
18. Methidathion Below Quantification
Limit 0.0050-
0.20mg/kg
19. Parathion Below
Quantification Limit 0.0050-
0.50mg/kg
20. Permethrin Below
Quantification Limit 0.0050-
1.0mg/kg
21. Perimiphos-methyl Below Quantification
Limit 0.0050-
4.0mg/kg
22. Fentrothion Below Quantification
Limit 0.0050-
0.50mg/kg
23. Cypermethrin and isomers Below Quantification
Limit 0.0050-
1.0mg/kg
24. Hexachlorocyclohexane Below
Quantification Limit 0.0050-
0.30mg/kg
25. Piperonyl-butoxide Below
Quantification Limit 0.0050-
3.0mg/kg
26. Heptachlor Below
Quantification Limit 0.0050-
0.050mg/kg
27. Fenvalerate Below
Quantification Limit 0.0050-
1.50mg/kg
28. Endosulfan Below
Quantification Limit 0.0050-
3.0mg/kg
Table 7

ACUTE ORAL TOXICITY TESTING OF HERBAL TEA COMPOSITION

[0066] An experiment was performed to evaluate acute oral toxicity of herbal tea composition in Wistar rats. The experiment was conducted as per OECD Guideline for Testing of Chemicals No. 423, December 17, 2001 (“Acute Oral Toxicity – Acute Toxic Class Method”. The Test Item i.e. Wistar rats are procured by Althea DRF, Dabur Research Foundation, 22, Site IV, Sahibabad, Ghaziabad-201010, Uttar Pradesh, India (CPCSEA registration No.: 1242/PO/RcBiBt/S/08/CPCSEA).

Test item Preparation:

[0067] 5 gm of the herbal teal composition was boiled in 20 ml water on hot plate for 10 min. The prepared decoction was filtered and cooled and was dose at a maximum 20 mL/kg of dose volume. The herbal tea composition was prepared fresh prior to oral administration.

Dose Administration:

[0068] Prior to dosing, each Wistar rat was kept on fasting overnight (approximately 15- 16 h) with water ad libitum. The test composition was administered orally in each Wistar rat based on the recent body weight to each Wistar rat,using stainless steel ball tipped intubation needle attached to sterile disposable syringe. The dose volume was maintained at 20 mL/kg body weight. The feed was provided between 3 to 4 hours of first post dosing.

[0069] Two sets, each containing six female Wistar rats, were formed to perform the experiment. Prior to dosing, the Wistar rats were kept on fasting overnight for upto 15-16 hours. The Wistar rats (1,2,3) in the first set were orally administered with adose of 250 mg/mL of the herbal tea composition and dose volume of 20 mL/kg twice a day (in a divided doses 10 mL/kg morning and 10 mL/kg evening).

Table 8 illustrates dosing concentration for both the sets of Wistar rats
Set Dose Concentration
(mg/mL) Sex No. of Animals per Step Animal No.
Set 1 250 Female 3 1-3
Set 2 250 Female 3 4-6
Table 8

[0070] No mortality was reported in the first set post 48 hours of dosing. Based on the survival pattern of the previously dosed Wistar rats after 48 hours, the Wistar rats (4,5,6) in the second set were treated with the confirmatory same dose of 250 mg/mL of the herbal tea composition.

Observation:

[0071] Clinical Signs – All Wistar rats were examined for clinical signs at 30 minutes and approximately at 1 hour, 2 hour, 3 hour, and 4 hour post dosing and later, once a day. This examination was repeated for 14 days

[0072] Mortality/Morbidity – All Wistar rats of both the sets were examined for mortality/morbidity, twice a day, daily, throughout the experimental period.

[0073] Body Weights – The body weight of all Wistar rats of both the sets was recorded on the day of receipt, and at time of animal selection. The percentage change in the body weight was calculated on day 0 (prior to dosing) to day 7, day 7 to 14 and day 0 to 14.

[0074] Gross Pathology- All the surviving Wistar rats at the time of termination i.e., on Day 14, were euthanized humanely by CO2 asphyxiation and gross pathological examination was performed for Wistar rats of set 1 and set 2.

Result:

[0075] Clinical Signs- No clinical signs of evident toxicity were observed in Wistar rats of first set and second set throughout the observation period.

Table 9 illustrates clinical signs in individual Wistar rat of first set.
Animal No. Day
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14
30
minutes 1h 2h 3h 4h
1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1
2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1
3 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1
1: Normal
Table 9

Table 10 illustrates clinical signs in individual Wistar rat of second set
Animal No. Day
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14
30
minutes 1h 2h 3h 4h
4 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1
5 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1
6 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1
1: Normal
Table 10

[0076] Mortality/Morbidity- No mortality/morbidity was observed in Wistar rats of first and second set.

Table 11 illustrates experimental results of mortality/morbidity testing in female Wistar rats of first and second set.

Set
Animal No. Day Mortality/Morbidity
Set 1
1 0 - 14 Nil
2 0 - 14 Nil
3 0 - 14 Nil
Set 2
4 0 - 14 Nil
5 0 - 14 Nil
6 0 - 14 Nil
Nil = No morbidity/mortality
Table 11

[0077] Body Weight- An increase in the body weight of all the Wistar rats of first and second set was reported on day 7, day 14 as compared to day 0.

Table 12 illustrates experimental results of body weight and dost volume in individual Wistar rats
Set Animal No. Body Weight (g) Body Weight (g) Percentage (%)
change in Body Weight
Day 0 Day 7 Day 14 Day 0-7 Day 7-14 Day 0-14
Set 1 1 178.32 183.83 188.20 3.09 2.38 5.54
2 174.56 189.51 200.22 8.56 5.65 14.70
3 205.91 213.54 215.48 3.71 0.91 4.65
Mean 186.26 195.63 201.30 5.12 2.98 8.30
SEM 9.88 9.11 7.89 1.73 1.40 3.21
Set 2
4 197.07 198.72 199.67 0.84 0.48 1.32
5 192.82 197.57 200.12 2.46 1.29 3.79
6 205.24 208.75 210.57 1.71 0.87 2.60
Mean 198.38 201.68 203.45 1.67 0.88 2.57
SEM 3.64 3.55 3.56 0.47 0.23 0.71
Table 12

[0078] Gross Pathology –No external or internal abnormality was reported in Wistar rats of first and second set.

Table 13 illustrates experimental results of gross pathological examination in female Wistar rats of first and second set

Set
Animal No. External Examination Internal Examination
Set 1 1 NAD NAD
2 NAD NAD
3 NAD NAD
Set 2 4 NAD NAD
5 NAD NAD
6 NAD NAD
NAD = No Abnormalities Detected
Table 13

[0079] Conclusion: It was reported that herbal tea composition, when administered in Wistar rats of first and second set at the dose of 250 mg/mL, and dose volume of 20 mL/kg twice a day (In a divided doses 10 mL/kg morning and 10 mL/kg evening), was considered to be safe for oral consumption. Further, the Wistar rats showed no mortality, no signs of toxicity and no gross pathological changes in a single day dosing.

[0080] Germ kill efficacy of the herbal composition: As shown in Figure 1, the time kill analysis may monitor the effect of various concentrations of the herbal composition over time in relation to the stages of the growth of the bacteria. The herbal composition showed >95% germ-kill efficacy against various pathogenic micro-organisms i.e., Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus epidermis, Listeria monocytogene, Candida albicans, Aspergillus brasilensis, Escherichia coli and Salmonella abony, at intervals of 30 and 60 seconds.

[0081] Anti-inflammatory activity of the herbal composition: The anti-inflammatory activity of the herbal composition was evaluated. A maximum inhibition of 81.7% in the level of cytokine (IL-8) in Lung cells (as shown in Figure 2a) and a maximum of 100% inhibition in the level of cytokine (IL-6) in Immune cells (as shown in Figure 2b) was reported, thus indicating anti-inflammatory activity of the herbal composition.

[0082] Analgesic potential of the herbal composition: As shown in Figure 3, the analgesic potential of the herbal composition was evaluated. The herbal composition showed a maximum of 87.8% inhibition in COX-2 enzyme activity as compared to enzyme control, thus indicating analgesic potential of the herbal composition.

[0083] Mucolytic potential of the herbal composition: The mucolytic potential of the herbal composition was evaluated. A maximum of 100% of inhibition of MUC5AC levels against TNF-alpha stimulation (as shown in Figure 4a) and a maximum of 79% decrease in the viscosity of egg white solution as compared to untreated sample (as shown in Figure 4b) was reported, thus indicating mucolytic potential of the herbal composition.

[0084] Antioxidant potential of the herbal composition: As shown in Figure 5, antioxidant potential of the herbal composition by scavenging stable free radical DPPH was evaluated. A maximum of 84% inhibition of free radical levels as compared to untreated sample was reported, thus indicating antioxidant potential of the herbal composition.

[0085] The advantages of the present invention are discussed herein:
• The herbal tea composition of the present invention is safe, natural/organic with negligible side effects.
• The herbal tea composition exhibits antibacterial, antiviral and anti-inflammatory properties, which relieves throat related infections and associated symptoms thereof.
• The herbal tea composition provides relief from cold, cough, sore throat, fever, flu and other throat related infections.

[0086] While this invention has been described in connection with what is presently considered to be the most practical and preferred embodiment, it is to be understood that the invention is not limited to the disclosed embodiments, but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the scope of the appended claims.
,CLAIMS:WE CLAIM:

1. A herbal tea composition, comprising:
• one or more herbal extracts in a range of 0.2% to 90% by weight of the total composition;
• one or more flavoring agents in a range of 0.1% to 5% by weight of the total composition;
• an active ingredient in a range of 1% to 80% by weight of the total composition;
• a bioenhancer in a range of 0.001% to 1% by weight of the total composition; and
• a solvent in a range of 1% to 10% by weight of the total herbal tea composition.

2. The herbal tea composition as claimed in claim 1, wherein the herbal extracts are selected from a group consisting of Withania somnifera (Ashwagandha), Ocimum basilicum (Basil), Panax ginseng (Asian ginseng), Ginkgo biloba (Ginkgo), Glycyrrhiza glabra (Licorice) and Zingiber officinale (Ginger) or a combination thereof.

3. The herbal tea composition as claimed in claim 1, wherein the flavoring agent is a plant based natural flavoring agents.

4. The herbal tea composition as claimed in claim 1, wherein the plant based natural flavoring agents are selected from a group of Basil flavor, Ashwagandha flavor, Orange flavor, Cinnamon flavor, Cardamom flavor, Anise flavor, Lemon flavor, Ginger flavor, Cardamom flavor, and Clove flavor, or a combination thereof.

5. The herbal tea composition as claimed in claim 1, wherein the active ingredient is in form of a ready-made hydroalcoholic extract containing parts of the herbs loaded with active ingredients.

6. The herbal tea composition as claimed in claim 1, wherein the parts of the herbs for preparing the hydroalcoholic extract are selected from a group consisting of fruit of Black Pepper (Piper nigrum), bark of Cinnamon (Cinnamomum zeylanicum), bud of Clove (Eugenia caryophyllus), bud of Cardamom (Elettaria cardamomum), leaf of tea plant (Camellia sinensis) and flower of Pomegranate (Punica granatum), or a combination thereof.

7. The herbal tea composition as claimed in claim 1, wherein the solvent is selected from a group consisting of Isopropyl alcohol (IPA), or ethyl alcohol.

8. A method for preparing the herbal tea composition as claimed in claim 1, comprising steps of:
a) adding herbal extracts in an apparatus;
b) straining the herbal extracts through a sieve;
c) granulating the extracts obtained in step (b) with a solvent and bioenhancers, resulting in the formation of granules;
d) allowing the granules to dry at a temperature in a range of 20? to 50? for 3 to 4 hours;
e) adding flavoring agents and the active ingredient to the granules obtained in step ; and
f) mixing the mixture obtained (d) thoroughly, forming a herbal composition.