DESC:Technical Field
The present disclosure relates to field of herbal composition. More particularly, the present disclosure relates to herbal composition and method of preparing herbal composition.
Background
The cause of morbidity in the current scenario among the world population is recognized as inflammatory diseases. For instance, the factors involved in the inflammatory process are present in many diseases like rheumatic arthritis, inflammatory bowel diseases, cancer, atherosclerosis, Asthma and allergy etc. Inflammation is a protective mechanism of the body, that includes vascular and cellular events to abolish or control the extent of injurious factors or the cause of tissue or cell injury by foreign invaders such as chemical, mechanical or thermal stimuli, microbial or infectious pathogens and normalizing the disturbed tissue homeostasis. Inflammation has two types: acute and chronic conditions. Acute inflammation begins rapidly, that could be a short-lived phenomenon involved in the overproduction of radicals, accumulation of fluids, plasma proteins, leukocytes, neutrophil infiltration, triggering the complex enzymes and release various inflammatory mediators, nitric oxide, cycloxygenase-2 etc.
The anti-pyretic, anti-inflammatory ayurvedic proprietary products in the market, has mostly time-tested classical recipes of Ayurveda which are traditionally used to combat fever and associated infections as one of the ingredients in the formulation. Classical ayurvedic products are blended with ayurvedic herbs or other formulations and proprietary ayurvedic products are developed. So, most of the products in the market have time-tested classical ayurvedic formulations in it.
Some examples of the currently available products in the market are FLU-O-CIL Tablet of Dhootpapeswar, Trishunn Tablet of Zandu, Malbet Tablet of Phyto, Pyrid tablet of Nagarjuna, Pyril DS tablets of Jammi Pharmaceuticals, Imvir Tablet by Phyto specialties, Fevnil -unity health care products, Febricurom Tablet, Cureom life science pirvate limited, Thane.Vepye tablet, vee remedies, Fever not Vostok herbals, Pyrant tablet ayursun pharma, Fever guard tablet from IMC.
The ayurvedic industry is precariously poised in the issues of herb availability and
sustainability. The wide acceptance of ayurvedic and herbal products has caused indiscriminate use of the herbs without considering their sustainability also. This has also caused profuse adulteration and admixture in the supply of herbs. Most of the classical formulations are polyherbal with herbs that are most difficult to source, which further increases the risk of admixture or adulteration and the quality of the final product. So, products with simple formulations with commonly available herbs that are cultivable are the need of the day. Simple formulations with easily available/cultivable herbs, ensure the quality and sustained use of the input materials. This also brings sustainability to manufacturing. Such simple herbs can be effectively combined and made into products that will deliver the intended results.
Methicillin-resistant Staphylococcus aureus (MRSA) is usually spread in the community by contact with infected people or things that are carrying the bacteria. Methicillin-resistant Staphylococcus aureus (MRSA) infection is caused by a type of staph bacteria that's become resistant to many of the antibiotics used to treat ordinary staph infections.
Therefore, there is a need to overcome the limitations associated with anti-pyretic, anti-inflammatory ayurvedic proprietary products.
Summary
In one aspect of the present disclosure, an herbal composition is provided. The herbal composition includes Adathoda vasica ranging from 10 to 20 % weight/weight (w/w) of the herbal composition, Androgarphis paniculata ranging from 10 to 20 % w/w of the herbal composition, Curcuma longa ranging from 10 to 20 % w/w of the herbal composition, Tinospora cordifolia ranging from 10 to 20 % w/w of the herbal composition, Oldendalia corymbosa ranging from 10 to 20 % w/w of the herbal composition, Zingiber officinale ranging from 10 to 20 % w/w of the herbal composition, and Carica papaya ranging from 10 to 20 % w/w of the herbal composition.
In some aspects of the present disclosure, the herbal composition is in the form of 100 mg active tablet.
In second aspects of the present disclosure, a method of preparing herbal composition is disclosed. The method includes washing and cleaning Adathoda vasica, Androgarphis paniculata, Curcuma longa, Tinospora cordifolia, Oldendalia corymbosa, Zingiber officinale and Carica papaya. The method further includes boiling Adathoda vasica, Androgarphis paniculata, Curcuma longa, Tinospora cordifolia, Oldendalia corymbosa, Zingiber officinale and Carica papaya with water at 80-120 degrees Celsius for 3 - 5 hours. The method further includes collecting extract from the boiled content. The method further includes repeating the boiling and extract collecting process for four times. The method further includes mixing the extracts together. The method further includes evaporating the mixed extracts to obtain a concentrated form of extracts. The method further includes powdering the concentrated form of extracts. The method further includes mixing the powdered extracts with one or more excipients. The method further includes punching the powdered extracts with one or more excipients into a tablet.
In some aspects of the present disclosure, the one or more excipients are selected from group includes one or more binders, one or more cosolvents, one or more fillers, one or more disintegrates, one or more lubricants, one or more surfactants, one or more emulsifying agents, one or more suspending agents, one or more sweeteners, one or more preservatives and combination thereof.
In some aspects of the present disclosure, the one or more binders are selected from group includes cellulose, methyl cellulose, polyvinyl pyrrolidine, Polyethylene glycol (PEG), gelatine, Polyvinylpyrrolidone (PVP), Hydroxypropyl methylcellulose (HPMC), sucrose, and starch.
In some aspects of the present disclosure, the one or more cosolvents are selected from group includes ethanol, propylene glycol, glycerine, glycofural, and polyethylene glycol (PEG).
In some aspects of the present disclosure, the one or more fillers are selected from the group includes lactose, sucrose, magnesium stearate, glucose, plant cellulose, calcium carbonate, and microcrystalline cellulose.
In some aspects of the present disclosure, the one or more sweeteners are selected from group includes sugar, sorbitol powder, maltitol and other sugar alcohols, sucralose, stevia, liquid glucose, and sorbitol.
In some aspects of the present disclosure, the one or more preservatives are selected from group includes the potassium sorbate, sodium benzoate, phenylmercuric nitrate, bronopol, propylene glycol, and benzylkonium chloride.
In some aspects of the present disclosure, one or more suspending agents are selected from group includes aqueous biological polymers, gum tragacanth, methylcellulose (MC), sodium carboxymethylcellulose (CMC), hydroxyethyl cellulose, and hydroxypropylmethylcellulose (HPMC).
Brief description of drawings
The accompanying drawings, which are incorporated in and constitute a part of this specification, show certain aspects of the subject matter disclosed herein and, together with the description, help explain some of the principles associated with the disclosed implementations. In the drawing,
Figure 1 illustrates a flowchart that depicts a method of preparing herbal composition, in accordance with an aspect of the present disclosure;
Figure 2 illustrate graph that depicts UV-Visible spectrum profile, in accordance with an aspect of the present disclosure;
Figure 3 illustrate graph that depicts FT-IR peaks, in accordance with an aspect of the present disclosure;
Figure 4 illustrate graph that depicts chromatogram of the herbal composition by GC-MS Spectrum, in accordance with an aspect of the present disclosure;
Figure 5 illustrate graph that depicts in vitro antioxidant activities by DPPH radical scavenging activity, reducing power (FRAP) and ABTS assay, in accordance with an aspect of the present disclosure;
Figure 6 illustrate graph that depicts the haematological analysis, in accordance with an aspect of the present disclosure;
Figure 7 illustrate graph that depicts determination of nitrite level, in accordance with an aspect of the present disclosure;
Figure 8 illustrate graph that depicts determination of PGE2 & COX-2 level, in accordance with an aspect of the present disclosure;
Figure 9 illustrate graph that depicts determination of lipid peroxidation level, in accordance with an aspect of the present disclosure;
Figure 10 illustrate graph that includes determination of MPO level in paw tissue, in accordance with an aspect of the present disclosure;
Figure 11 illustrate graph that depicts effect of herbal composition on inflammatory mediators in paw tissue, in accordance with an aspect of the present disclosure;
Figure 12 illustrate graph that depicts effect of herbal composition on antioxidant activity, in accordance with an aspect of the present disclosure; and
Figure 13 illustrate graph that depicts flow cytometric analysis of CD3+, CD4+ and CD8+, in accordance with an aspect of the present disclosure.
Detailed description of the preferred embodiments
Various embodiments of the disclosure are discussed in detail below. While specific implementations are discussed, it should be understood that this is done for illustration purposes only. A person skilled in the relevant art will recognize that other components and configurations may be used without parting from the spirit and scope of the disclosure. Thus, the following description and drawings are illustrative and are not to be construed as limiting. Numerous specific details are described to provide a thorough understanding of the disclosure. However, in certain instances, known details are not described in order to avoid obscuring the description.
References to one or an embodiment in the present disclosure can be references to the same embodiment or any embodiment; and, such references mean at least one of the embodiments.
Reference to "one embodiment", "an embodiment", “one aspect”, “some aspects”, “an aspect” means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the disclosure. The appearances of the phrase "in one embodiment" in various places in the specification are not necessarily all referring to the same embodiment, nor are separate or alternative embodiments mutually exclusive of other embodiments. Moreover, various features are described which may be exhibited by some embodiments and not by others.
The terms used in this specification generally have their ordinary meanings in the art, within the context of the disclosure, and in the specific context where each term is used. Alternative language and synonyms may be used for any one or more of the terms discussed herein, and no special significance should be placed upon whether or not a term is elaborated or discussed herein. In some cases, synonyms for certain terms are provided.
A recital of one or more synonyms does not exclude the use of other synonyms.
The use of examples anywhere in this specification including examples of any terms discussed herein is illustrative only, and is not intended to further limit the scope and meaning of the disclosure or of any example term. Likewise, the disclosure is not limited to various embodiments given in this specification. Without intent to limit the scope of the disclosure, examples of instruments, apparatus, methods and their related results according to the embodiments of the present disclosure are given below. Note that titles or subtitles may be used in the examples for convenience of a reader, which in no way should limit the scope of the disclosure. Unless otherwise defined, technical and scientific terms used herein have the meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. In the case of conflict, the present document, including definitions will control.
Additional features and advantages of the disclosure will be set forth in the description which follows, and in part will be obvious from the description, or can be learned by practice of the herein disclosed principles. The features and advantages of the disclosure can be realized and obtained by means of the instruments and combinations particularly pointed out in the appended claims. These and other features of the disclosure will become more fully apparent from the following description and appended claims, or can be learned by the practice of the principles set forth herein.
As mentioned above, there is a need to overcome the limitations associated with anti-pyretic, anti-inflammatory ayurvedic proprietary products. Therefore, the present disclosure provides an herbal composition and method of preparing herbal composition.
In some aspects of the present disclosure, herbal composition may be disclosed. The herbal composition may include Adathoda vasica ranging from 10 to 20 % weight/weight (w/w) of the herbal composition, Androgarphis paniculata ranging from 10 to 20 % w/w of the herbal composition, Curcuma longa ranging from 10 to 20 % w/w of the herbal composition, Tinospora cordifolia ranging from 10 to 20 % w/w of the herbal composition, Oldendalia corymbosa ranging from 10 to 20 % w/w of the herbal composition, Zingiber officinale ranging from 10 to 20 % w/w of the herbal composition and Carica papaya ranging from 10 to 20 % w/w of the herbal composition.
In some aspect of the present disclosure, the herbal composition may include Adathoda vasica ranging from 14.285 % weight/weight (w/w) of the herbal composition, Androgarphis paniculata ranging from 14.285 % w/w of the herbal composition, Curcuma longa ranging from 14.285 % w/w of the herbal composition, Tinospora cordifolia ranging from 14.285 % w/w of the herbal composition, Oldendalia corymbosa ranging from 14.285 % w/w of the herbal composition, Zingiber officinale ranging from 14.285 % w/w of the herbal composition and Carica papaya ranging from 14.285 % w/w of the herbal composition.
In some aspects of the present disclosure, the herbal composition may be in the form of 100 mg active tablet.
Figure 1 illustrates a flowchart that depicts a method 100 of preparing herbal composition, in accordance with an aspect of the present disclosure. The method 100 of preparing herbal composition may include the following steps:
At step 102, Adathoda vasica, Androgarphis paniculata, Curcuma longa, Tinospora cordifolia, Oldendalia corymbosa, Zingiber officinale and Carica papaya may be washed and cleaned;
At step 104, washed Adathoda vasica, Androgarphis paniculata, Curcuma longa, Tinospora cordifolia, Oldendalia corymbosa, Zingiber officinale and Carica papaya may be boiled with water at 80-120 degrees Celsius for 3 - 5 hours;
At step 106, extract obtained from step (104) may be collected;
At step 108, the step (104) and step (106) may be repeated for four times;
At step 110, the extracts obtained from step (108) may be mixed;
At step 112, the mixed extracts obtained from step (110) may be evaporated to get a concentrated form of extracts;
At step 114, the concentrated form of extracts obtained from step (112) may be powdered;
At step 116, the powdered extracts obtained from step (114) may be mixed with one or more excipients; and
At step 118, the powdered extracts with one or more excipients obtained from step (116) may be punched into a tablet.
In some aspects of the present disclosure, the one or more excipients may be selected from group that includes one or more binders, one or more cosolvents, one or more fillers, one or more disintegrates, one or more lubricants, one or more surfactants, one or more emulsifying agents, one or more suspending agents, one or more sweeteners, one or more preservatives and combination thereof.
In some aspects of the present disclosure, the one or more binders may be selected from group that includes cellulose, methyl cellulose, polyvinyl pyrrolidine, Polyethylene glycol (PEG), gelatine, Polyvinylpyrrolidone (PVP), Hydroxypropyl methylcellulose (HPMC), sucrose, starch and the like.
In some aspects of the present disclosure, the one or more cosolvents may be selected from group that includes ethanol, propylene glycol, glycerine, glycofural, polyethylene glycol (PEG) and the like.
In some aspects of the present disclosure, the one or more filler may be selected from the group that includes lactose, sucrose, magnesium stearate, glucose, plant cellulose, calcium carbonate, microcrystalline cellulose and the like.
In some aspects of the present disclosure, the one or more sweeteners may be selected from group that includes sugar, sorbitol powder, maltitol and other sugar alcohols, sucralose, stevia, liquid glucose, sorbitol and the like.
In some aspects of the present disclosure, one or more preservatives may be selected from group that includes the potassium sorbate, sodium benzoate, phenylmercuric nitrate, bronopol, propylene glycol, benzylkonium chloride and the like.
In some aspects of the present disclosure, one or more suspending agents may be selected from group that includes aqueous biological polymers, gum tragacanth, methylcellulose (MC), sodium carboxymethylcellulose (CMC), hydroxyethyl cellulose, hydroxypropylmethylcellulose (HPMC) and the like.
EXPERIMENTAL STUDIES AND RESULTS
I. CHEMICAL CHARACTERIZATION OF THE HERBAL COMPOSITION
In one exemplary scenario, the chemical characterization of herbal composition was analyzed by way of UV-VIS spectrum analysis, FT-IR spectroscopy analysis, and GC-MS analysis.
Figure 2 illustrate graph that depicts UV-Visible spectrum profile, in accordance with an aspect of the present disclosure.
In one exemplary scenario, the qualitative UV-VIS spectrum profile of the herbal composition was selected at a wavelength from 200 to 1022.5 nm due to the sharpness of the peaks and proper baseline. The UV-VIS spectrum profile showed the peak at 202 nm with maximum absorption of 0.7501 as shown in Figure 2.
Figure 3 illustrate graph that depicts FT-IR peaks, in accordance with an aspect of the present disclosure.
Peak
value Vibration Functional group
3421 O-H Stretching Alcohol
1633 C=O and NH vibrations Amide
1414 C-H Stretching Aromatic
1385 C-H deformation vibration Alkyl
1152 C–O–C stretching Ether
1080 PO2 vibrations Phosphodiester
1023 C-O-C vibrations Ether
Table 1. FT-IR peak values and functional groups present in herbal composition.

In another exemplary scenario, to characterize the functional group present in the herbal composition, Thermo-fisher Scientific’s Fourier Transform Infrared spectroscopy model Nicolet iS50 FT-IR operating in transmission mode at a resolution of 4 cm-1 is used. When the herbal composition was passed into the FT-IR, the functional groups of the herbal components were separated based on the ratio of their peak. Seven bands at 3421, 1633, 1414, 1385, 1152, 1080, and 1023 cm-1 have appeared in the n FTIR spectrum of the herbal composition as shown in Figure 3. The FT-IR gave a broad peak at 3421 cm-1 which indicated the presence of O-H stretching. It gave a strong peak at 1633 cm-1 which indicated the presence of C=O and NH vibrations and showed amide groups, 1414 cm-1 attributed to C-H Stretching having aromatic functional groups, the peak around 1023 and 1152 cm-1 are due to ether group, 1385 cm-1 C-H deformation vibration having Alkyl group, 1080 cm-1 PO2 vibrations of Phosphodiester group. The FT-IR spectrum confirmed the presence of alcohols, aliphatic, carboxylic acids, and esters. The results of FT-IR peak values and functional groups were represented in Table 1.
Figure 4 illustrate graph that depicts chromatogram of the herbal composition by GC-MS Spectrum, in accordance with an aspect of the present disclosure.
In another exemplary scenario, the chemical constituents of the herbal composition were identified by using GC-MS analysis as shown in figure 4.
II. INVITRO ANTIOXIDANT EFFECT OF THE HERBAL COMPOSITION
Methodology
In another exemplary scenario, Invitro antioxidant activity was evaluated using DPPH, ABTS and FRAP assay. The free radical scavenging ability of the herbal composition was tested by DPPH radical scavenging assay as described by Blois and Desmarchelier et al. (Blois 1958; Desmarchelier et al. 1997). The FRAP assay was determined using the potassium ferricyanide–ferric chloride method and the ABTS radical scavenging assay was calculated on the basis of the method.
Results
Figure 5 illustrate graph that depicts in vitro antioxidant activities by DPPH radical scavenging activity, reducing power (FRAP) and ABTS assay, in accordance with an aspect of the present disclosure.
In another exemplary scenario, free radical scavenging is the major attribute of all the antioxidants. the antioxidant activity of the herbal composition against DPPH and ABTS radicals and FRAP are investigated. Figure 5 demonstrated the dose response curve of DPPH, ABTS and FRAP. From the result, it was observed that the hydrogen donating ability of the herbal composition to neutralize the DPPH radical was lower at lower concentration, but it was increased with increase in concentration. Similarly, in the case of ABTS radical scavenging, the herbal composition showed dose dependency, where the highest antioxidant activity was found at 100 µg/mL concentration. The examination of antioxidant potential through the ferric reduction assay exhibit that there was an increase in the absorbance as the concentration of the sample increases which implies that the herbal composition possesses strong antioxidant activity in a dose-dependent manner.
III. ANTI-INFLAMMATORY EFFECT OF THE HERBAL COMPOSITION
Methodology
Carrageenan induced paw edema model
In another exemplary scenario, a dose-response study for the anti-inflammatory activity of the herbal composition against carrageenan-induced paw edema model was carried out in the range of 50-400 mg/kg body weight. From this, a dose of 100 mg /kg bwt was found to be the minimal concentration for maximal edema inhibition. Hence the minimal effective dose of 100 mg/kg bwt was selected for further evaluation of the anti-inflammatory effect of the herbal composition.
Treatment protocol and experimental design for acute inflammation.
In another exemplary scenario, anti-inflammatory activity was measured using carrageenan-induced rat paw edema assay by Winter et al. 1962. Male Wistar rats were randomly divided into four groups with six rats in each.
Group 1: Normal Control (0.5% DMSO in normal saline), (NC)
Group 2: Carrageenan, (Carr)
Group 3: Carrageenan + herbal composition (100 mg/kg bwt), (Carr + FB)
Group 4: Carrageenan + Diclofenac (20mg/kg bwt), (Carr + Dic)
Diclofenac dose (20mg/kg bwt) was used for a standard anti-inflammatory drug study. The herbal composition and standard drug diclofenac were given intra-peritoneally, followed by injection of 1% carrageenan suspension in 0.9% NaCl solution after 1 hr into the sub-plantar tissue of the right hind paw. Paw volume was measured at hourly intervals for 3hrs by using a paw edema meter. Anti-inflammatory activity was measured as the percentage reduction in edema level when the extract was compared to control. Activity = 100 – (100 x average drug-treated /average for control). At the end of the third hour, animals were sacrificed by euthanasia. Blood and paw tissue were collected for various biochemical analyses. Paws from experimental rats were amputated above the ankle, degloved, and snap-frozen in liquid nitrogen. Frozen paws were pulverized in a liquid nitrogen bath and divided into aliquots for extraction and analysis of inflammatory markers such as Nitrite level, PGE2 level, TBARS, COX-2, IL-1ß, IL-6, TNF-a, MPO activities and hematological analysis such as C-reactive protein (CRP) and white blood cell (WBC).
Results
Inhibitory effect of the herbal composition on carrageenan-induced paw edema rats (dose-response study)
SL. NO Drug concentration (mg/kg of body weight) Percentage of inhibition (%) at 3rd hour
Diclofenac 20 15
The herbal composition
50
55 %
100 88.8 %
200 88.6%
400 88.5 %
Table.2. Effect of the herbal composition on carrageenan-induced paw edema model (dose-response study).
In another exemplary scenario, to investigate, the effect of the herbal composition on anti-inflammatory activity was evaluated using the carrageenan-induced paw edema model. The subcutaneous administration of carrageenan generated an increase in the paw size in rats due to edema and the highest peak was seen after 3rd hour of the experiment in the carr induced group indicating an acute paw inflammation. Table 2 shows the effect of the herbal composition and standard drug as compared to carrageenan control at 3rd hr in carr induced animals. Carr induced animals were treated with different concentrations of herbal composition (50, 100, 200, and 400 mg/kg bwt). The experimental data of the herbal composition administrated showed a significant decrease in the paw edema and our findings revealed that at a dose of 100 mg/kg.bwt prevented paw edema with a percentage of inhibition of 88.8% at 3rd hr in the treated rats when compared to the control group. From this result, the herbal composition showed a minimal dose with a potent inhibitory effect on the carr group when compared to the Standard drug (Diclofenac, 20 mg/kg.bwt). Hence, the herbal composition can act as a good anti-inflammatory agent.
Haematological analysis- CRP and WBC
Figure 6 illustrate graph that depicts the haematological analysis, in accordance with an aspect of the present disclosure.
In another exemplary scenario, as a result of inflammation induced by the injection of Carr, haematological parameters WBC count and CRP level were raised significantly (p=.05) in the Carr control group; but this was considerably suppressed by the supplementation of the herbal composition in comparison with standard drug and Carr treated rats as shown in Figure 6.
Effect of the herbal composition on the Nitric oxide Level
Figure 7 illustrate graph that depicts determination of nitrite level, in accordance with an aspect of the present disclosure.
In another exemplary scenario, as shown in Figure 7, injection of carr into the rat hind paw induced a marked increase in the serum NO level 3 hrs after injection. The treatment of rats with the herbal composition (100 mg/kg) caused a significant (p=.05) reduction of increased NO generation by carrageenan in serum as compared with the standard drug and carrageenan control group.
Effect of the herbal composition on the Prostaglandin E2 (PGE2) and COX-2 Level
Figure 8 illustrate graph that depicts determination of PGE2 & COX-2 level, in accordance with an aspect of the present disclosure.
In another exemplary scenario, a significant anti-inflammatory activity of the herbal composition was exhibited by inhibiting PG production. The result in Figure 8 showed that the PGE2 level increased significantly (p=.05) in the paw tissue 3 hrs after carrageenan injection. The treatment of rats with the herbal composition (100 mg/kg) and diclofenac (10 mg/kg, i.p.) caused a significant (p=.05) reduction of increased PGE2 level by carrageenan in paw tissue as shown in Figure 8. However, the effect of the herbal composition in paw tissue was more potent than that of standard drug and carrageenan induced group. Moreover, increased COX-2 level in carr administrated group was controlled by the herbal composition supplementation as shown in Figure 8.
Effects of the herbal composition on Lipid peroxidation level
Figure 9 illustrate graph that depicts determination of lipid peroxidation level, in accordance with an aspect of the present disclosure.
In another exemplary scenario, there was a significant (p=.05) increase in TBARS level in the Carr control group, resulting in high lipid peroxidation process. In contrast, the rats treated with the herbal composition showed a significant (p=.05) decrease in the level of TBARS as compared to the Carr 10 and Dic treated group as shown in Figure 9.
Effects of the herbal composition on Myeloperoxidase enzyme accumulation
Figure 10 illustrate graph that includes determination of MPO level in paw tissue, in accordance with an aspect of the present disclosure.
In another exemplary scenario, administration of the herbal composition significantly (p=.05) reduced the neutrophil content in carr treated group, evaluated from myeloperoxidase activity. The recruitment of neutrophils in the paw of the carr group was higher than normal control, whereas the herbal composition treatment drastically reduced the myeloperoxidase concentration, thereby inhibiting the neutrophil recruitment as compared to standard drug and carr control group as shown in Figure 10.
Effects of the herbal composition on cytokine secretion
Figure 11 illustrate graph that depicts effect of herbal composition on inflammatory mediators in paw tissue, in accordance with an aspect of the present disclosure.
In another exemplary scenario, to further elucidate the mechanism of action of the herbal composition in inflammation, proinflammatory cytokines such as TNF-a, IL-6 and IL-1ß were determined using ELISA technique. The paw tissue level of TNF- a in the carr-induced inflammation group was significantly (p=.05) increased compared with the normal control group, but treatment with the herbal composition reduced its level significantly (p=.05) which is comparable with the diclofenac treated group as shown in Figure 11. As shown in Figure 11, the carr group showed a significantly (p=.05) increased level of IL-1ß, whereas treatment with the herbal composition showed significant (p=.05) inhibition in tissue IL-1ß level when compared with standard drug and carrageenan control. Also, IL-6 levels increased dramatically in the carr control whereas the herbal composition caused a significant (p=.05) decrease in IL-6 levels while compared with the carr control group. Similarly, comparing the standard drug treatment group dramatically decreased the IL-6 level. The results were shown in Figure 11. These data imply that the herbal composition acts as an inhibitor of carr induced inflammation in paw tissues by decreasing haematological markers, inflammatory mediators like NO, PGE2, MPO, TBARS and pro-inflammatory cytokines such as TNF-a, and IL-1ß production in vivo.
IV. IMMUNOMODULATORY ACTION OF THE HERBAL COMPOSITION
Methodology
In another exemplary scenario, cyclophosphamide induced experimental animal model was used for evaluating the immunomodulatory effect of the herbal composition.
Treatment protocol and experimental design for Immunomodulation study.
In another exemplary scenario, animal treatment as follows:
Group 1- Normal control (no treatment) (NC)
Group 2 – cyclophosphamide + herbal composition (CP+FB) (100mg/kg body weight/day)
Group 3 –cyclophosphamide (CP) (10 mg/ kg body weight/day)
Group 4 –cyclophosphamide + levamisole standard drug (CP+STD) (50mg / kg body weight/day)
After the experimental duration of 21 days, antioxidant activity of SOD, CAT, GPx and GSH was evaluated and also to find out the presence of T-lymphocyte subsets CD3, CD4 and CD8-alpha in animal using flow cytometer.
Results
Antioxidant activity of the herbal composition
Figure 12 illustrate graph that depicts effect of herbal composition on antioxidant activity, in accordance with an aspect of the present disclosure.
In another exemplary scenario, the antioxidant activities of the enzymatic antioxidants SOD, CAT, GPx and the non-enzymatic antioxidant GSH respectively represented in Figure 12. The results showed that there was a significant reduction in these antioxidants in cyclophosphamide induced animals when compared with the normal control rats. However, the activity of these antioxidants was significantly increased after the administration of the herbal composition. As evident from the results, the herbal composition is more efficient than the standard drug in enhancing the antioxidant activities.
In another exemplary scenario, Figure 12 shows an effect of the herbal composition on the antioxidant activity. ‘a’-Statistical difference with normal group at P = 0.05. ‘b’-Statistical difference with diabetic treated rats at P < 0.05. SOD: U- enzyme concentration required to inhibit chromogen production by 50% in 1 min/mg protein. Catalase: U-µmolH2O2 decomposed/min/ mg protein. GPx: U-µmol 5 NADPH oxidized / min/ mg protein. GSH: U- µmol/g tissue.
Flow cytometry analysis
Figure 13 illustrate graph that depicts flow cytometric analysis of CD3+, CD4+ and CD8+, in accordance with an aspect of the present disclosure.
In another exemplary scenario, compared to the normal control group, the percentages of CD3+, CD4+ and CD8+ T cells in the Cyclophosphamide rat decreased significantly, that shows rat’s cellular immune function was inhibited. As compared to the Cyclophosphamide group, the percentages of CD3+ CD4+ and CD8+ T cells and the ratio of CD3+ /CD4+/CD8+ in the herbal composition treated group increased significantly as shown Figure 13, which implies that the herbal composition could increase the level of CD3 + /CD4+/CD8+ T cell, and directly enhances the cellular immune function in the immunosuppressed animals.
Advantages:
• The present disclosure provides anti-inflammatory, antioxidant, antimicrobial, and immunomodulatory products.
• The present disclosure provides an herbal composition that combats fever, and body pain and enhances immunity in immunocompromised condition.
• The present disclosure provides an herbal composition that counter the viral fevers, and manage the inflammation associated with these fevers thereby resolving the associated issues like secondary infections that occur in the respiratory system, body aches, headaches, and modulating immunity so that further susceptibility to diseases is prevented.
• The present disclosure provides an herbal composition that fighting respiratory affections often associated with pyrexia.
• The present disclosure provides an herbal composition that combats a superbug and Methicillin resistant Staphylococcus aureus (MRSA).

The implementation set forth in the foregoing description do not represent all implementations consistent with the subject matter described herein. Instead, they are merely some examples consistent with aspects related to the described subject matter. Although a few variations have been described in detain above, other modifications or additions are possible. In particular, further features and/or variations can be provided in addition to those set forth herein. For example, the implementation described can be directed to various combinations and sub combinations of the disclosed features and/or combinations and sub combinations of the several further features disclosed above. In addition, the logic flows depicted in the accompany figures and/or described herein do not necessarily require the particular order shown, or sequential order, to achieve desirable results. Other implementations may be within the scope of the following claims. ,CLAIMS:I/We claim (s):
1. An herbal composition comprising:
Adathoda vasica ranging from 10 to 20 % weight/weight (w/w) of
the herbal composition;
Androgarphis paniculata ranging from 10 to 20 % w/w of the
herbal composition;
Curcuma longa ranging from 10 to 20 % w/w of the herbal
composition;
Tinospora cordifolia ranging from 10 to 20 % w/w of the herbal
composition;
Oldendalia corymbosa ranging from 10 to 20 % w/w of the herbal
composition;
Zingiber officinale ranging from 10 to 20 % w/w of the herbal
composition; and
Carica papaya ranging from 10 to 20 % w/w of the herbal
composition.

2. The herbal composition as claimed in claim 1, wherein the herbal composition is in the form of 100 mg active tablet.

3. A method (100) of preparing herbal composition comprising:
washing and cleaning (102) Adathoda vasica, Androgarphis paniculata, Curcuma longa, Tinospora cordifolia, Oldendalia corymbosa, Zingiber officinale and Carica papaya;
boiling (104) washed Adathoda vasica, Androgarphis paniculata, Curcuma longa, Tinospora cordifolia, Oldendalia corymbosa, Zingiber officinale and Carica papaya with water at 80-120 degrees Celsius for 3 - 5 hours;
collecting (106) extract obtained from step (104);
repeating (108) the step (104) and step (106) for four times;
mixing (110) the extracts obtained from step (108);
evaporating (112) the mixed extracts obtained from step (110) to get a concentrated form of extracts;
powdering (114) the concentrated form of extracts obtained from step (112);
mixing (116) the powdered extracts obtained from step (114) with one or more excipients; and
punching (118) the powdered extracts with one or more excipients obtained from step (116) into a tablet.

4. The method (100) of preparing herbal composition as claimed in claim 3, wherein the one or more excipients are selected from group comprising one or more binders, one or more cosolvents, one or more fillers, one or more disintegrates, one or more lubricants, one or more surfactants, one or more emulsifying agents, one or more suspending agents, one or more sweeteners, one or more preservatives and combination thereof.

5. The method (100) of preparing herbal composition as claimed in claim 4, wherein the one or more binders are selected from group comprising cellulose, methyl cellulose, polyvinyl pyrrolidine, Polyethylene glycol (PEG), gelatine, Polyvinylpyrrolidone (PVP), Hydroxypropyl methylcellulose (HPMC), sucrose, and starch.

6. The method (100) of preparing herbal composition as claimed in claim 4, wherein the one or more cosolvents are selected from group comprising ethanol, propylene glycol, glycerine, glycofural, and polyethylene glycol (PEG).

7. The method (100) of preparing herbal composition as claimed in claim 4, wherein the one or more fillers are selected from the group comprising lactose, sucrose, magnesium stearate, glucose, plant cellulose, calcium carbonate, and microcrystalline cellulose.

8. The method (100) of preparing herbal composition as claimed in claim 4, wherein the one or more sweeteners are selected from group comprising sugar, sorbitol powder, maltitol and other sugar alcohols, sucralose, stevia, liquid glucose, sorbitol.

9. The method (100) of preparing herbal composition as claimed in claim 4, wherein the one or more preservatives are selected from group comprising the potassium sorbate, sodium benzoate, phenylmercuric nitrate, bronopol, propylene glycol, and benzylkonium chloride.

10. The method (100) of preparing herbal composition as claimed in claim 4, wherein the one or more suspending agents are selected from group comprising aqueous biological polymers, gum tragacanth, methylcellulose (MC), sodium carboxymethylcellulose (CMC), hydroxyethyl cellulose, and hydroxypropylmethylcellulose (HPMC).