ALLOPOLYHERBAL GEL FORMULATION AGAINST MICROORGANISM: IN-VITRO STUDIES

ALLOPOLYHERBAL GEL FORMULATION AGAINST MICROORGANISM: IN-VITRO STUDIES

Laxmi Srivastava, J. N. Mishra, D. K. Vishwakarma * and Chitranshu Pandey

Kailash Institute of Pharmacy & Management, Gida, Gorakhpur, Uttar Pradesh, India.

ABSTRACT: The cases of skin infections are increasing daily, and the conventional treatment loses its strength in treating those skin infections because of resistance of the microorganisms against the medications, including antibacterial as well as antifungal with the topical and oral administration, or both. The advancement of the treatment of treating skin infections by the combination with the synthetic drug (allopathic medicine) with the polyherbal formulation is known as Allopolyherbal formulation, which gives a wonderful result in the treatment of skin infections due to microorganisms (bacteria & fungus) by lowering the side effects and increasing the pharmacological activity of the synthetic drug (allopathic medicine) with the synergistic therapy in targeting the microbes and also shows the better patient compliance with the reliable therapeutic effects. In this research work, the allopolyherbal formulation contains the polyherbal formulation, which includes A. indica (Neem), O. tenuiflorum (Shyamatulsi), P. emblica (Amla), and A. barbadensis (Aloe) which have the potency enriched with antimicrobial activity and the allopathic drug, the antibacterial (chloramphenicol) and antifungal (terconazole) are used to treat many of the skin infections caused by S. aureus, P. aeruginosa, C. albicans, and A. niger. The overall resulted formulation of allopolyherbal gel as antibacterial (F4) and antifungal (F4) with PHF (B) shows better eradication rate as compared to all formulation and also the further evaluation such as physicochemical as well as in-vitro studies with stability was performed for particular formulation.

Keywords: Skin infection, Allopolyherbal formulation, Allopathic medicine, Polyherbal therapy, Patient compliance, Eradication rate

INTRODUCTION: Allopolyherbal plan is described as the definition which consolidates the mix of two exceptional specifying, for instance, the allopathic definition (designed drug) and polyherbal plan; the inventive idea of incorporating the allopathic with normal is to adjust everything from the ordinary treatment for the help of patient and to degree the past therapeutics of the prescription for the high-level world.

Other than this, in the relationship between's the allopathic drug and the allopolyherbal plans, which have a comparable effect on the medicine anyway, the optional impact is overpowered by the decline in the piece of the made prescription.

Polyherbal Formulation: Polyherbal plan is described as the blend of more than one zest use with the helpful preparation for treating sicknesses. The possibility of polyherbal definition is found is ayurvedic and other standard medicine systems.

In 1300 A.D. the thought was polyherbalism was writing in "Sarangdhar Samhita" in which plant definitions and joined concentrates of plants are picked rather than individual ones.

Due to synergism, polyherbalism bound a couple of benefits which isn't open in single local definition. On a very basic level, this structure is promptly used to treat various disorders, including diabetes.

TABLE 1: EXAMPLE OF POLYHERBAL FORMULATION

Background of Allopathic and Ayurveda Medicine System:

Allopathic Medicine: Allopathic is derived from two words they are “Allos” which means opposite and “Pathos” means suffering. In 1810, Samuel Hanneman designed the practice of medicine and termed as allopathy, which is opposed to homeopathy. Allopathic medicines were defined by WHO in 2001 as the “Broad category of medicine practice i.e., some time called biomedicine western medicine evidence based medicine or modern medicine”. The treatment with allopathic medicine is also known as “Conventional heroic medicine”.

Ayurveda: Ayurveda is an ancient traditional system of treatment, which was 2500 years ago, that balance the soul, body, and brain additionally expects to incorporate. This equilibrium is accepted that lead to bliss, and well-being forestalls disease.

In Ayurveda, the combinations of panchamah-bhutas is known as Tridoshas, which are

1. Vata (Vayu and Akash)
2. Pitta (Agni and Fire)
3. Kapha (Prithvi and Apa).

Whereas Ayurveda incorporates the usage of normally gotten flavors, for instance, flavors, minerals, flavors, intervention, mental tidiness, smells, sound, yoga, practice, and mechano-designs to get out the start liberated from the infection by improvement harmony. They made solid areas for a to stop their occurrence of lopsidedness. The key tendency is that flimsiness between tridosha causes debasements while astounding prospering is achieved when there is perseverance between these fundamentals. The reality behind Ayurveda is to vexed miserable repulsiveness and to happen with a tranquil flourishing life.

Advancement in Medicine System: Here, the advancement in the medicine system refers to the modification of allopathic medicine with ayurvedic medicine i.e., a single herb or in the herb-herb combination (Polyherbal Formulation) is required now a day just because the increased cost of medicine, as well as their side effect, has become a great task when the public health is concerned. Polyherbal definition started to obtain a noticeable quality all over the planet, due to the way that polyherbal plan gains a couple of benefits which isn't thwarted in allopathic drugs.

Topical Administration: A skin solution is a medication that applies to a particular put on or in the body. Generally, the skin association of only applied to the body surface, for instance, skin or mucous film to treat through tremendous extent of semi-solid portion structure, for instance, Creams, Foams, Gels, Lotions and Ointment.

Gels are homogenous, semi-solid preparation usually consisting of solution or dispersion of one or more medicaments in suitable hydrophilic or hydrophobic bases as per I.P.

FIG. 1: SCHEMATIC PATHWAY OF DRUG MECHANISM VIA SKIN

The above figure shows how drugs enter the skin and the therapeutic action. The action is produced inside the skin by two pathways Fig. 1.

Transcellular Pathway: Drugs in this lane go directly into skin through the phospholipid layer and the dead keratinocytes (cytoplasm) that contain the layer corneum.

Intracellular Pathway: In this part, the medicine helps in the skin into spaces of the skin's cells, by making the course extra scrambled.

FIG. 2: TRANSCELLULAR AND INTRACELLULAR PATHWAY OF DRUG ABSORPTION

MATERIAL AND METHOD:

Collection of Plant Material: Collection of specimens of herb A. indica, P. emblica, O. tenuiflorum, A. barbadensis were collected from near M.R.D. life sciences Pvt. Ltd. Lucknow, and authenticated by CSIR -CIMAP Lucknow U.P.

Chemical: The drug Chloramphenicol is taken as the market preparation (Starphenicol CADILA pharmaceutical Ltd.) and the Terconazole is the gift sample for the M.R.D. life sciences Pvt. Ltd. Carbapo l934 and 940, starch, sodiumalginate, gumtragacanth, polyethyleneglycol, pro-pyleneglycol, Ethylenediamine tetraacetic acid (EDTA) and triethanolamine (TEA) were obtained from Himedia laboratories Pvt. Ltd. Mumbai. P. Ether, Methanol, Ethanol, Chloroform, Acetone, Dimethyl sulfoxide (DMSO) were purchased from Avantor Performance Materials India Ltd.

Methodology:

Preparation of Extract: Table 2 the gathered new leaves of A. indica, A. barbadensis, and new consequences of P. emblica and entire plant of O. tenuiflorum were washed in water to make them liberated from dust and new material and dried in hot air stove at 40 °C to stay away from debasement of phytoconstituents, straight forwardly following drying the plant materials were coarsely powder with the assistance of mortar pestle and blend processor and kept in impenetrable fixed holder Six dissolvable which were Acetone, Chloroform, Ethyl acidic destructive induction, Methanol, P. Ether, refined water were utilized in moderate cold maceration system in the level of 1:10 (Extract: Solvent) ²².

Phytochemical Analysis: The 50% of methanolic and ethanolic removes were prepared to test the phytoconstituents in the flavors as alkaloid, flavonoid, carb, amino destructive, cardiovascular glycoside, phenol, terpenoid, tannin, saponin, steroid according to the standard show in Table 3.

Preparation of Gel: The gel was ready by the Fusion strategy Fig: 3A and 3B in which different gelling specialists were utilized, and after that, the polyherbal extricates were blended in with the suitable measure of medication and the other excipients like EDTA, Triethanolamine, propylene glycol were additionally consolidated in that as support, additive and pH change Table 4A and 4B.

Evaluation:

Physicochemical Property of Allopolyherbal Gel Table: 5A & 5B):

Physical Appearance: Physical parameters such as; colour, appearance, and consistency were checked visually.

pH: Calibrated digital pH meter is used to measure the water solution (one percent) of the formulation at a constant temperature.

Viscosity: Shaft # C 50-1 (speed of 50 rpm at room temperature) and the affirmation of consistency was done in a three-over lay with the consistency of their organized regular gel enumerating was assessed by using Brookfield Viscometer (Brookfield Engineering Laboratories, USA).

Spreadability: Two arrangements of glass slides were taken with the standard lookout; two slides contain Polyherbal itemizing between them of length 60mm. The stuck excess gel had been taken out on the external layer of the glass slides and put on a stand with no disrupting impact. The time it took to improve the upper slide to a distance of 60 mm was measured with a 20 g weight, and the time it took to improve the top slide to that distance was influenced by the weight. The spreadability was resolved by utilizing the going with recipe, which was not fixed in stone, by repeating the evaluation numerous times.

Spreadibility = (Weight × Length) ̸ Time

Homogeneity: After the definitions have been sat in a compartment, all made subtleties were pursued for homogeneity by visual assessment.

Antibacterial Study: Changed agar well scattering method was used to affirm antibacterial activity of different definitions (Graph 1).

This procedure developed supplement agar plates with 0.2ml of stock culture of minute life forms (S. aureus and P. aeruginosa). For 1 hour, the plates were left to dry. A perfect 8 mm drill was utilized to cut 3 equal-distance wells in all of the panels; 1g of Allopolyherbal gels plan was taken using, Chloramphenicol medicine was used as a control sample, and refined freshwater was employed as an awful control. At 370°C, the plates were tortured for 24 hours. Antibacterial activity was determined by measuring the breadth of impediment zones (in mm).

Antifungal Activity: Sabouraud dextrose agar well dispersal strategy was used to affirm antifungal activity of different enumerating (Graph 1). In this procedure, sabouraud dextrose agar plates were developed with 0.2ml of stock culture of parasite (C. albicans and A. niger). For 1 hour, the plates were left to dry. In all of the plates, a pristine 8 mm drill was used to cut 3 equidistance wells; 1g of Allopolyherbal gel solution was collected with it. The medication terconazole was utilized as a control sample and the water sample was used as a bad control, both of which were thrown into the wells with abandon. At 370°C, the plates were tortured for 24 hours. The antifungal properties were discovered by evaluating the size of the barrier zones (in mm).

Colony Factor Unit (CFU): The CFU was resolute by the readiness of fluid stock for microbes (Nutrient stock) and for the organism (Sabouraud Dextrose stock) was ready in the adequate amount which had the option to move in seven test tubes (9 ml), and one test tube is marked as clear with the microorganisms (20 µl) just, and the rest test tube was added with the 20 µl of the microbes, and in various test tubes the allopolyherbal gel definitions (10 mg) were added and kept all the test tubes in the shaker hatchery at 37 °C for 24 hrs., after that the above-pre-arranged stocks (20 µl) were spread on the supplement agar media (microscopic organisms) and sabouraud dextrose agar media (growth) and afterward kept in the hatchery at 37 °C for  24 hrs and the CFU was seen by advanced settlement counter in Table 6.

In-vitro Drug Release Study: In-vitro appearance of the drug was performed by egg cell film strategy, where the egg cell layer is used as the normal film.

Extraction of Egg Membrane: Void the egg with internal filling material and plunged in 1N HCl for 24 hrs, and after that, the film was gained and retained methanol before used approx. for 2 hrs.

Permeability Study: Ventured through a test chamber and break from its closed side and the egg layer was appended there with the help of string and put the estimating glass stacked up with phosphate support 7.4 put on the appealing stirrer with alluring touch, by and by the test tube with egg film was placed in the phosphate support 7.4 and the Allopolyherbal Gel Formulation (10 µl) was filled, and the temperature will be 37± 0.50C at 50 rpm. Eliminated the model (1 ml) at a specific period of minutes and added the phosphate support in the estimating utencil to stay aware of the sink condition, and repeated it for 5 extra times, and the absorbance was taken against 278 nm (Chloramphenicol) and 220 nm (Terconazole) by UV evident spectrophotometer (Graph 3).

Stability Study: The last itemizing, for instance, allopolyherbal gel, was presented to strength peruses up for a range of one month at various temperature and dampness conditions. 25±2 °C/60% ±5% RH, 40±2 °C/75% ±5% RH and refrigeration temperature (4 °C). The models were attempted and evaluated right away and a while later on the fifth day, the fifteenth day, and 30^th day from the day of commencement of the definitions Table 7A and 7B.

RESULTS:

Extraction of Herbs: The extraction process of herbs involved cold maceration with different solvents, but the best results were shown in a few solvents at different percentages Table 2.

TABLE 2: EXTRACTIONS OF HERBS WITH THEIR ZONE OF INHIBITIONS

Phytochemical Analysis: The phytoconstituents analysis follows the standard protocol Table 3.

TABLE 3: PHYTOCHEMICAL SCREENING OF HERBS, MEOH = METHANOLIC EXTRACT, ETOH = ETHANOLIC EXTRACT, PRESENT (+) AND ABSENT (-)

Formulation Table: The formulation of allopolyherbal gel was prepared using the following ingredients in Table 4A & 4B.

TABLE 4A: ANTIBACTERIAL FORMULATION OF ALLOPOLYHERBAL GEL

TABLE 4B: ANTIFUNGAL FORMULATION OF ALLOPOLYHERBAL GEL

Physicochemical Evaluation of Allopolyherbal Gel Formulations

TABLE 5A: PHYSICOCHEMICAL CHARACTERISTICS OF ANTIBACTERIAL ALLOPOLYHERBAL GEL

TABLE 5(B): PHYSICO CHEMICAL CHARACTERISTICS OF ANTIBACTERIAL ALLOPOLYHERBAL GEL

Antimicrobial Sensitivity Test: In-vitro antimicrobial sensitivity test was done on the mentioned pathogens with the formulated allopolyherbal gel graph 1 (A) and 1 (B).

GRAPH 1: AST OF ALLOPOLYHERBAL GEL FORMULATIONS

Colony Factor Unit (CFU): By determining the CFU the final formulation of antibacterial (F4) and antifungal (F4) of the allopolyherbal gel was determined in Table 6 and Graph 2.

TABLE 6: CFU OF ANTIBACTERIAL & ANTIFUNGAL ALLOPOLYHERBAL GEL FORMULATIONS

GRAPH 2: CFU OF ANTIBACTERIAL & ANTIFUNGAL ALLOPOLYHERBAL GEL FORMULATIONS

FIG. 3A: ANTIBACTERIAL ALLOPOLYHERBAL GEL (F4)                                                              

FIG. 3B: ANTIFUNGAL ALLOPOLYHERBAL GEL (F4)

In-vitro Drug Release: In-vitro drug release of the allopolyherbal gel formulation was conducted by diffusion flask (egg membrane). The result of the drug release is shown in Graph 3.

GRAPH 3: CUMM % DRUG RELEASE OF ALLOPOLYHERBAL GEL FORMULATIONS

Stability Study: The study of stability was done only on the formulation (F4) of antibacterial and antifungal, which showed the best results by the above-mentioned results.

TABLE 7A: STABILITY STUDY OF ANTIBACTERIAL ALLOPOLYHERBAL GEL FORMULATION (F4)

TABLE 7B: STABILITY STUDY OF ANTIFUNGAL ALLOPOLYHERBAL GEL FORMULATION (F4)

CONCLUSION: The repeat of ingesting the allopathic medications to treat an impressive parcel of the defilements achieved by the S. aureus, P. aeruginosa, C. albican and A. niger which results to convey unpleasant reactions. Consequently, the normal fixes are considered as safeguarded as the designed ones, and the meanings of flavors with single substance and polyherbal structures are fostering the market demand all over the planet. Assurance of the plants with their ideal obsessions is basic to convey the best supportive results. Local plants such as A. indica (Neem), O. tenuiflorum (Shyamatulsi), P. emblica (Amla) and A. barbadensis (Aloe) are selected and incorporated into the different gel bases as a non-oily (aqueous) topical polyherbal formulations which also contain the marketed preparations in the very less concentration and quantity. This research concludes that the formulation of allopolyherbal gel, formulated against the microbes as an antibacterial and an antifungal, had shown wonderful results in eradicating the microbes with good permeability and stability studies; the formulations are also best able against specific conditions.

ACKNOWLEDGEMENT: I am very grateful to acknowledge my institution, i.e., Kailash Institute of Pharmacy and Management, which has allowed me to perform this research work and equally to the MRD Life Sciences P. Ltd.

CONFLICTS OF INTEREST: There is no conflict of interest related to this research work.

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    How to cite this article:

    Srivastava L, Mishra JN, Vishwakarma DK and Pandey C: Allopolyherbal gel formulation against microorganism: in-vitro studies. Int J Pharm Sci & Res 2023; 14(2): 867-75. doi: 10.13040/IJPSR.0975-8232.14(2).867-75.

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