STANDARDIZATION OF AMALAKYADI CHURNA: A POLY HERBAL FORMULATION
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1 Page5156 Indo American Journal of Pharmaceutical Research, 2016 ISSN NO: STANDARDIZATION OF AMALAKYADI CHURNA: A POLY HERBAL FORMULATION Neha Soni, Pankaj Pradhan *, Yogesh Sharma, Darsh Gautam Himachal Institute of Pharmaceutical Education & Research, Nadaun, Himachal Pradesh, India. ARTICLE INFO Article history Received 04/04/2016 Available online 30/04/2016 Keywords Amalakyadi Churna, Physicochemical Parameter, Phytochemical Analysis, Antimicrobial Activity. Corresponding author Neha Soni Himachal Institute of Pharmaceutical Education & Research, Nadaun, Himachal Pradesh pnkj.pradhan@gmail.com ABSTRACT In the few decades, there has been exponential growth in the field of herbal medicines. Most of the traditional system of medicine is effective but they lack standardization. So there is a need to develop a standardization technique. Standardization of herbal formulation is essential in order to assess the quality, purity, safety and efficacy of the drug. Pharmaceutical ayurvedic research is aimed at meeting the medical needs of the population for whom appropriate therapeutic remedies are not available or at those that are available are but not effective for various disorders. While meeting medical needs of a polyherbal formulation set some parameters to ensure that the formulation shows desired pharmacological action against various diseases. The selection of an appropriate drug should take into account apart from medical needs and innovative potential for success. The standardization of crude drug materials includes authentication, organoleptic evaluation, ash values, extractive values, moisture content determination and Carr s index etc. These parameters are required for authentication of any herbal drug and its formulation. Please cite this article in press as Neha Soni et al. Standardization of Amalakyadi Churna: A Poly Herbal Formulation. Indo American Journal of Pharmaceutical Research.2016:6(04). Copy right 2016 This is an Open Access article distributed under the terms of the Indo American journal of Pharmaceutical Research, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
2 Page5157 INTRODUCTION Ayurveda means the science of life. Since ancient time a variety of pharmaceutical dosage form have been used in Ayurvedic system of medicine and some of them are in practice even today. [1] Ayurveda is a time-tested, trusted worldwide plant based system of medicines and consists of various Ayurvedic formulations such as Asava, Arishta, Ghrita, Taila, Churna, Kwatha and much more. Among which churna is a fine powder consisting of a single drug or a mixture of more than one drug in a dry form. [2] The World Health Organization (WHO) has appreciated the importance of medicinal plants for public health care in developing nations and has evolved guidelines to support the member states in their efforts to formulate national policies on traditional medicine and to study their potential usefulness including evaluation, safety and efficacy. [3] Inspite of the large number of Ayurvedic formulation available in the market, for many of them, Standard for their quality is yet to be laid. [4] Various marketed formulation shows dose variation, content variation and lack of standardization which affect its therapeutic activity, therefore it is imperative to establishment of quality control parameters for herbal of Ayurvedic formulations which will be in alignment with modern technology [5]. Standardization is an essential factor for polyherbal formulation in order to assess the quality of drugs based on the concentration of their active principle. It is very important to establish a system of standardization for every plant medicine in the market, since the scope of variation in different batches of medicine is enormous. Plant material when used in bulk quantity may vary in its chemical content and therefore, in its therapeutic effect according to different batches of collection e.g. collection in different season and/or collection from sites with different environmental surrounding or geographical location. The increasing demand of the population and chronic shortage of authentic raw materials have made it incumbent, so there should be some sort of uniformity in the manufacture of Ayurvedic medicines so as to ensure quality control and quality assurance [6]. The quality control of herbal crude drug & formulation is important in justifying their acceptability in modern system of medicines. Standardization of synthetic drugs offers no problem with very well defined parameters of analysis. It is not uncommon to have as many as five or more different herbal ingredients in one single formulation. The batch to batch variation starts from the collection of the raw materials itself in absence of any reference standard for identification. WHO has emphasized the need to ensure quality control of medicinal plants products by using modern techniques and by applying suitable standards and parameters. Standardized products and services are valuable. User confidence builder s being perceived as: 1. Safe 2. Healthy 3. Secure 4. High Quality 5. Flexible Standardization brings important benefits to business including a solid foundation upon which to develop new technologies and an opportunity to share and enhance existing practices. Standardization also plays a pivotal role in assisting Governments, Administrations, Regulators and the legal profession as legislation, regulation and policy initiatives are all supported by standardization. [7] Churna is defined as a fine powder of drug or drugs in ayurvedic system of medicine. Drugs mentioned in patha, are cleaned properly, dried thoroughly, pulverized and then sieved. The churna is free flowing and retains its potency for one year, if preserved in an air tight container. Triphala churna, Trikatu churna, Drakeshadi churna and Sudharsana churna are some of examples. These forms of medicament are prescribed generally because of their particle size. Smaller the particle size greater is the absorption rate from GIT and hence the greater is bioavailability. It is prescribed by the Ayurvedic physician for treating conditions such as diabetes, indigestion, constipation etc. Preparation of polyherbal medicine is based on traditional methods in accordance with the procedure given. Due to lack of modern phramacopoeial standards laid down and followed processing of herbal formulation using traditional methods, the medicine prepare may not have the desired quality and batch-to-batch consistency. Hence there is a need for standardization of herbal formulation as prescribed by WHO. It is the process of evaluating the quality and purity of crude drugs by organoleptic character, physiochemical parameters and flow properties of powder. [8] Amalakyadi churna is one of the famous polyherbal ayurvedic churna formulation which is useful in Cough, phthisis, pain, wrinkles in the skin, diseases of skin, disease of abdomen, fistula-in-anus, dysuria, chronic obstructive jaundice, disorder due to vatta aggravation, disorders of pitta aggravation, piles, and disorder due to kapha aggravation [9]. The formulation was stored in well closed airtight container in dry and cool place. Physicochemical & Phytochemical screening studies have not been reported for the formulations. With this aim the current project was designed to prepare and standardized the Amalakyadi churna in accordance with the WHO guidelines. MATERIALS & METHODS Plant material The crude drugs used in preparation of Amalakyadi churna were collected from local Market of Nadaun in January All plant parts were then dried in shade, powdered and passed through sieve no. 60 and lastly packed in a well closed container to protect them from moisture. Each ingredients 5gm weight separately, mixed together to obtain a homogeneous blend. [9]
3 Page5158 Table 1: Composition of Formulation. S. No. Ingredient Latin Name Part Quantity 1 Amla Phyallanthus emblica Fruit 5 gm 2 Citraka Plumbago zeylanica Root 5 gm 3 Pathya Terminalia Chebula Fruit 5 gm 4 Pippali Piper longum Fruit 5 gm 5 Saindhava Lava Rock Salt gm Organoleptic Evaluation Organoleptic evaluation refers to evaluation of formulation by color, odour, taste, texture etc. The organoleptic characters of the samples were carried out based on the method mentioned in API. [10] Physicochemical Evaluation Analysis of Physicochemical Constants of In-house formulations has been done to evaluate the quality and purity of the powder drug. In physicochemical evaluation, ash value such as total ash, acid insoluble ash was evaluated. The ash value indicates the presence of inorganic salts present in the drug. The water soluble and alcohol soluble extractive values were determined. The information collected from this evaluation was useful for standardization and obtaining the quality standards for crude drugs as well as for formulations. Determinations of these physicochemical constants were done as per procedures mentioned in accordance with WHO guidelines. [11] Physical Characterization Physical characteristics of In-house formulation were done as per pharmacopoeial procedures. The physical characteristics like moisture content, Bulk density, Tap density, Angle of repose, Hausner s ratio and Carr's index indicates the flow properties as well as interparticulate resistance between powders. The information collected from this evaluation was crucial to avoid ambiguous predictions of stability or solubility of formulation. [12] Extraction: About 250 gm of dry churna was taken in a closed bottle and it was defatted with Petroleum ether. The deffating was continued for 9-10 days with occasional shaking. The Petroleum ether extract was filtered. The marc left after Petroleum ether deffating was taken out and dried under shade to get a dry mass, then extracted with Methanol and water (hydro alcoholic) by using hot percolation extraction. The extraction was continued for 72 hours. The hydro alcoholic extract was concentrated under reduced pressure to a semisolid mass and was made free from solvent. The final obtained extract was weighed; percentage yield was calculated and stored in a cool place. [13] Antimicrobial Activity Microorganism Four Reference Bacteria, Vic. Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 29213, Pseudomonas aeruginosa ATTCC and Bacillus cereus ATCC 6633 were used during the present study and were obtained from Dr. B. Lal Institute, Jaipur, Rajasthan. The bacteria were grown in nutrient broth at 37 0 C and maintained on nutrient agar slants at 4 0 C. Antibacterial Assay Muller- Hinton Agar plates are prepared by pouring ml of the medium in to each sterilized Petridis and are allowed to set at room temperature. The cell suspension is inoculated over the surface of agar medium using sterile cotton swab. The two cups are scooped in each plate using a sterile cork borer of 8 mm diameter. Then the solution of test compounds (25 μl, 50 μl, 100 μl and control 100 μl) are added in cups by using micropipettes and these plates are incubated at 37 0 C for 48 hr. Standard drug ciprofloxacin (5 mcg) is used. The Zone of inhibition is measured in mm for each organism. [14] RESULT & DISCUSSION In house formulation was prepared in accordance with the Ayurvedic formulary of India. As part of standardization procedure, the finished product Amalakyadi churna was tested for relevant physical & chemical parameters with antimicrobial activity. Organoleptic Evaluation The colour, odour and taste of the formulation were evaluated manually and given in Table No. 2
4 Page5159 Table 2. Organoleptic Descriptions of Amalakyadi churna. S. No. Parameters Observation 1 Colour Light Brown 2 Odour Characteristic 3 Taste Bitter 4 Texture Smooth Physicochemical Evaluation Quality tests for different Amalakyadi churna and its individual ingredients were performed for moisture content, ash content, water soluble extractive, methanol soluble extractive, acid insoluble ash and water insoluble ash were found to be within standard ranges. Various physiochemical parameters of churna is given in Table 3. Variations were observed in most of the physicochemical parameters studied. Table 3. Physiochemical Descriptions of Amalakyadi churna. S. No. Parameters Observation 1 LOD % 3.7± Crude Fiber 0.19± Total Ash 18.5± Acid Insoluble Ash 1.0± Water Soluble Ash 12± Alcohol Extractive Value 33.5± Water Soluble extractive Value 20.4 ±0.17 Physical characterization The result of Physical Parameter of Amalakyadi churna was tabulated in Table 4. Table 4. Physical Characters of Amalakyadi churna. S. No. Parameters Observation 1 Bulk Density 0.38± Tapped Density 0.56± Angle of Repose Hausner s Ratio 1.47± Carr s Index 27.52± Porosity % 86±0.018 Antimicrobial Activity Hydro alcoholic extract is test for antibacterial activity. The doses of 1000 mg /ml of extracts were made by dissolving appropriate quantity of extracts in DMSO. Standard drug Ciprofloxacin is used. The solution of test compounds (25 μl, 50 μl, 100 μl and control 50 μl) are added in cups by using micropipettes and these plates are incubated at 37 0 C for 48 hr. The zone of inhibition is measured in mm for each organism. Controls with DMSO did not show any activity. The crude extract shows positive antimicrobial activity against both gram positive and negative bacteria (table 5, figure 1, 2, 3, 4.) Table 5: Showing Effect of Amalakyadi churna extract on microbial growth. S. No. Drug Microorganism Amalakyadi Churna Extract(25 μl) Amalakyadi Churna Extract (50 μl) Amalakyadi Churna Extract (100 μl) Ciprofloxacin (50 μl) 1 Escherichia coli 3 mm 7 mm 9 mm 34 mm 2 Staphylococcus aureus 4 mm 11 mm 12 mm 23 mm 3 Pseudomonas aeruginosa 6 mm 12 mm 15 mm 29 mm 4 Bacillus cereus 7 mm 13 mm 18 mm 31 mm
5 Page5160 Fig. 1: Zone of Inhibition against Escherichia coli. Fig. 2: Zone of Inhibition against Staphylococcus aureus. Fig. 3: Zone of Inhibition against Pseudomonas aeruginosa. Fig. 4: Zone of Inhibition against Bacillus cereus. CONCLUSION From the present investigation various standardization parameters such as Organoleptic Standards, Physical Standards, Physicochemical standards and safety evaluation were carried out, it can be concluded that the formulation of Amalakyadi churna was in accordance with the standards laid down for churna. The study shows that the contents of formulation presents within the permissible limits as per WHO, all these investigations are not specified in the standard literature such as in pharmacopoeia, which could helpful in authentication of Amalakyadi churna. The result of present study will may be serve as reference monograph in the preparation of drug formulation. The present investigation clearly demonstrates the significant antibacterial activity of Hydro alcoholic extract of Amalakyadi churna against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Bacillus cereus in vitro. These results indicate the potential use of this formulation in management of bacterial diseases caused by Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Bacillus cereus. This study tends to express that the Amalakyadi churna have active ingredients against these gram positive and negative bacteria. ACKNOWLEDGEMENT The authors are thankful to HIPER, Nadaun & Dr. B. Lal Lab, Jaipur for providing necessary facilities to carry out the research work. The authors are also thankful to the Mr. Ashok Sharma, Honorable Chairman, Himachal Institute of Pharmaceutical Education & Research, Nadaun & Dr. Parshuram Rai, Principal, HIPER, and Nadaun for providing necessary facilities and financial support for this study.
6 Page5161 REFERENCE 1. Jarald EE. Textbook of pharmacognosy and phytochemistry. 1st ed. CBS publication, Delhi 2007; Indian Pharmacopoeia-Vol III. Controller of Publications, Ghaziabad 2007; World Health Organization (WHO). Quality Control Methods for Medicinal Plant Materials. AITBS Publishers, Delhi 2002; Tekeshwar K., Chandrashekar K.S., Tripathi D.K., Nagori K., Pure S., Agrawal S., Tamsil J.A. Standardisation of Gokshuradi churna-an ayurvedic polyherbal formulation. J Chem Pharm Res. 2011; 3(3): Gupta V., Jain U.K. Establishment of quality standards for herbal formulation, sitopaladi churna. Int J Drug For Res. 2011; 2 (1): Karthi J., Kalvimoorthi V., Thamizh Mozhi M. Standardisation of Sudharshana Churna Polyherbal Formulation. IJPCBS. 2012; 2(3): Bahuguna Y., Zaidi S, Kumar N., Rawat K. Standardization of Polyherbal Marketed Formulation Triphala Churna. RRJPP. 2014; 2(3): Chamoli N, Agarwal K., Saini P. Preparation and Standardization of Madhushoonya Churna: Polyherbal Formulation. International Journal of Innovative Pharmaceutical Research. 2013; 4(1): Ayurvedic Formulary of India. Part I, 2nd ed. Controller of Publications, New Delhi 2003; Siddiqui and Hakim MA. Format for the pharmacopoeial analytical standards of compound formulation, workshop on standardization of Unani drugs, (appendix), January, Central Council for Research in Unani Medi cine (CCRUM), New Delhi, Pradhan P., Joseph L., George M., Kaushik N., Chulet R. Pharmacognostic, Phytochemical & Quantitative Investigation of Saraca asoca leaves. Journal of Pharmacy Research. 2010; 3(4): Surisetty RR, Xavier FG, Seetharaman S, Duraipandian C. Standardization of Marketed Churna an Ayurvedic Polyherbal Formulation. Int. J. Pharm. Sci. Rev. Res. 2014; 28(2): Yadav Yashraj, Mohanty PK, Kasture SB. Anti-inflammatory activity of hydro alcoholic extract of Quisqualis indica Linn. flower in rats. Int. J. of Pharm. & Life Sci. 2011; 2(8): Pradhan P., Joseph L., George M., Chulet R. Antibacterial Activity of Saraca Asoca Leaves Extract. THE PHARMA REVIEW. 2010; 2(3)
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