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Table of Contents
ORIGINAL ARTICLE
Year : 2020  |  Volume : 8  |  Issue : 2  |  Page : 114-121

Study of Panchashirishadi Agad on the basis of physicochemical and phytochemical analysis


Department of Agadtantra Vyavhar Ayurved Evum Vidhi Vaidhayak, Mahatma Gandhi Ayurved College Hospital and Research Centre, Datta Meghe Institute of Medical Sciences (Deemed to be University), Wardha, Maharashtra, India

Date of Submission29-Feb-2020
Date of Decision30-Mar-2020
Date of Acceptance08-May-2020
Date of Web Publication07-Sep-2020

Correspondence Address:
Dr. Shanti SK Manyala
C/O Anil Mahajan, Rashtrabhasha Road, Rathi Layout, Wardha, Maharashtra.
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/JISM.JISM_10_20

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  Abstract 

Background: Many antidotes are mentioned in the classical text for the management of animal poisoning. Panchashirishadi Agad is one of the formulations indicated for Kita Visha (insect bite). Aim: To assess the physicochemical and phytochemical properties of Panchashirishadi Agad.Setting and Design: This was an original analytical study. Materials and Methods: Five parts of Shirish (Albizia lebbeck) were collected from local area of Wardha (Sewagram) and Panchashirishadi Agad was formulated as per the reference of classical text. Result: After the analytical study, the physicochemical properties that include loss on drying at 105°C, total ash value, acid-insoluble ash, water-soluble extractive, alcohol-soluble extractive, pH, and particle size were analyzed and the obtained values were 12.0%, 10.0%, 1.0%, 11.0%, 13.0%, 5.45%, and 100 mesh, respectively. In the similar way, the herbal formulation is devoid of microbial contamination. Phytochemical analysis includes the presence of 48% polyphenols. Conclusion: As the formulation contains more percentage of polyphenols, it can be anticipated that it can act as antiallergic agent.

Keywords: Agada, Ayurveda, Keeta, Samhita


How to cite this article:
Manyala SS, Chalakh SP. Study of Panchashirishadi Agad on the basis of physicochemical and phytochemical analysis. J Indian Sys Medicine 2020;8:114-21

How to cite this URL:
Manyala SS, Chalakh SP. Study of Panchashirishadi Agad on the basis of physicochemical and phytochemical analysis. J Indian Sys Medicine [serial online] 2020 [cited 2020 Oct 30];8:114-21. Available from: https://www.joinsysmed.com/text.asp?2020/8/2/114/294428




  Introduction Top


Agad is “antidote,” which counteracts the hazardous effects of poisons on living beings.1 According to ancient classical texts, certain Agada are broad acting and some others are target specific in action. Agada act as a specific antidote for various types of poisoning.2

Panchashirishadi Agad is one of the potent formulations mentioned in Shushrut Samhita, which is indicated for Kita Visha (insect poison).3Kita Damsha (insect bite) and Kita Visha lakshanas (insect poison symptoms) are very common in tropical country like India. Deaths are very rare in cases of insect bites such as honeybee sting and wasp sting. Mortality can occur when there is any “anaphylactic reaction” associated with it, which depends on the severity and frequency of bite.

Generally people do not report hospitals for insect bites. People follow certain home remedies or take certain antihistaminics like “cetirizine” from over the counter in pharmacies. Even if victims report to hospitals, they are prescribed topical and systemic antihistaminics and corticosteroids. There is very less awareness in public regarding the untoward effects of these antihistaminics and corticosteroids.4

These medications only give temporary relief to the patient, but they leave many untoward effects on the body system. On the contrary, Ayurvedic formulations focus on the cause, rather than the symptoms and treat the cause accordingly.

In the Samhita, we find reference of Panchashirishadi Agad as a very potent formulation for Kita Visha (insect poison).5 No any previous work was performed to study the efficacy of this compound. Thus, this study is very fruitful to establish the potency and standard parameters of Panchashirishadi Agad on the analytical basis.

This study aimed at assessment of the physicochemical and phytochemical analysis of Panchashirishadi Agad.


  Materials and Methods Top


Raw materials required: Five parts of Albizia lebbeck (roots, flower buds, leafs, seeds, bark) and Trikatu were required for the study.

Authentication of drugs: Identification and authentication of five parts of A. lebbeck (leafs, roots, seeds, flower buds, and bark) was performed at the Foundation for Revitalisation of Local Health Traditions (FRLHT), Bengaluru. Zingiber officinale, Piper nigrum, Piper longum, and rock salt were identified and authenticated from Department of Dravyaguna, Mahatma Gandhi Ayurved College Hospital and Research Centre, Salod (H), Wardha. The Institutional Ethical Committee approval was obtained on April 9, 2018.

Procedure of Preparation of Drug

All the raw materials were properly cleaned, dried, and were taken in equal quantity. All the ingredients (five parts of A. lebbeck) were properly trichurated and mixed together and Panchashirishadi AgadChurna was prepared. 10 g of Panchashirishadi Agad Churna was taken and Kwath was prepared. Later Ghan was prepared and Trikatu was added as Prakshep dravya. Finally, Panchashirishadi Ghan Vati was prepared. Ghan Vati was dried in shade and stored in air tight container. Panchashirishadi Agad Churna and Vati were physicochemically and phytochemically analyzed.


  Observation and Result Top


Observations of Panchashirishadi Agad Ingredients

The ingredients required for the preparation of Panchashirishadi Agad formulation were the five parts of A. lebbeck, Z. officinalis, P. nigrum, P. longum, and rock salt. All the crude ingredients were taken in equal quantity (50 g each). After the sorting and cleaning process, each ingredient was powdered into fine Churna form. The quantity of the ingredients after powdering was A. lebbeck flower bud powder: 37.22 g, A. lebbeck leaves powder: 38.42 g, A. lebbeck bark powder: 41.15 g, A. lebbeck seeds powder: 38.14 g, A. lebbeck root powder: 24.15 g, Z. officinalis powder: 42.14 g, P. nigrum powder: 41.13 g, and P. longum powder: 43.12 g.

During the cleaning, pounding, and sieving process of all the ingredients, there was loss of some portion of the ingredients. The weight loss of each ingredient was A. lebbeck flower bud powder: 12.78 g, A. lebbeck leaves powder: 11.58 g, A. lebbeck bark powder: 8.85 g, A. lebbeck seeds powder: 11.86 g, A. lebbeck root powder: 25.85 g, Z. officinalis powder: 7.86 g, P. nigrum powder: 8.87 g, and P. longum powder: 6.88 g. The percentage weight loss of each ingredient was A. lebbeck flower bud powder: 25.56%. The quantity taken for Panchashirishadi Agad preparation in powder form was 5 g each of the powder of the five parts of A. lebbeck. The quantity taken for Panchashirishadi Agad preparation in the tablet form was 10 g powder of each of the five parts of A. lebbeck and 4 g powder of Z. officinalis, P. nigrum, and P. longum. All these observations are shown in [Table 1].
Table 1: Observation of Panchashirishadi Agad ingredients

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Pharmaceutical Observation of Panchashirishadi Agad (Churna)

The quantity of each ingredient required for the preparation of Panchashirishadi Agad formulation was 5 g. The total quantity of ingredients required for the preparation of the formulation is 25 g and total quantity of formulation prepared is 25 g. These observations are seen in [Table 2].
Table 2: Pharmaceutical observation of Panchashirishadi Agad (Churna)

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Pharmaceutical Observations of Panchashirishadi Agad (Vati)

The total quantity of ingredients taken for the preparation of Panchashirishadi AgadVati was 62 g. All the ingredients were heated at 60°C–70°C for approximately 40 min to obtain 400 mL Kwath. The obtained quantity of Kwath dravya was 162mg and the total quantity of Prakshep dravya added was 12 g. The total quantity of weight gain in gram was 100 g and percentage weight gain was 38.27%. These observations are mentioned in [Table 3].
Table 3: Pharmaceutical observation of Panchashirishadi

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Organoleptic Characters of Panchashirishadi Agad (Churna and Vati)

The color of Panchashirishadi Agad in Churna (powder form) was yellow and in Vati (tablet form) was buff color. The odor of both Churna and Vati was noncharacteristic. The taste of both Churna and Vati was Tikta-Katu and both Churna and Vati form was soft in touch. All these observations are shown in [Table 4].
Table 4: Organoleptic characters of Panchashirishadi Agad

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Physicochemical Analysis and Microbiological Specifications of Panchashirishadi Agad (Churna and Vati form)

The loss on drying at 105°C in Churna form was 12% and Vati form was 2.5%. The total ash value of Churna form was 10% and Vati form was 7.5%. The acid-insoluble ash of Churna form was 1% and Vati form was 2.5%. The water-soluble extractive value of Churna was 11% and Vati was 34%. The alcohol-soluble extractive value of Churna was 13% and Vati was 20.0%. The pH of Churna was 5.45 and Vati was 5.60. The particle size of Churna was 100 mesh and Vati was 60 mesh. All the observations are tabulated in [Table 5].
Table 5: Analysis of Panchashirishadi Agada ( quantity of sample taken––10 g). Analytical observation of Panchashirishadi Agada (Churna) and (Vati)

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In the microbiological specifications, the total viable count, Enterobacteriaceae, total fungus count, Escherichia coli, Salmonella, Staphylococcus aureus, and Pseudomonas aeruginosa were absent in both the Churna and Vati form. Thus, the formulation in both Churna and Vati form was microbial contamination free. All these observations are mentioned in [Table 5].


  Discussion Top


Panchashirishadi Agad is one of the polyherbal formulations mentioned in Shushrut Samhita, Kalpa Sthana, which is indicated for Kita Visha (insect poison). In this study, Panchashirishadi Agad was analyzed from physicochemical and phytochemical aspect. The acid-insoluble ash value of Panchashirishadi Agad in Churna and Vati form was 1% and 2.5%, which indicated that the presence of inorganic matter in the formulation was within the prescribed limit in Ayurvedic Pharmacopiea of India (API) with reference to A. lebbeck.

In a similar way, the total ash value of Panchashirishadi Agad in Churna and Vati form were 10% and 7.5%, respectively, which was within the prescribed limit. It indicates that the presence of silica and calcium oxalate content in the formulation was in the prescribed limit with reference to API.

Loss on drying of Panchashirishadi Agad in Churna and Vati form was 12% and 2.5%, which is suggestive that the compound is not likely to get contaminated by fungal growth. The physical parameter such as pH was determined to confirm its acidic or basic nature and it was observed that the pH of Panchashirishadi Agad in Churna and Vati form has pH 5.45 and 5.60, respectively, which indicates that it is basic in nature.[6] Basic nature of this formulation indicates that it does not have any adverse effects on the gastrointestinal tract.

During preparation of Panchashirishadi Agad, aseptic conditions and sterilized instruments were used. Furthermore, microbial studies have been performed, which revealed the absence of Enterobacteriaceae and fungal growth including E. coli, Salmonella, S. aureus, and P. aeruginosa.

High-performance thin layer liquid chromatography (HPTLC) analysis of Panchashirishadi Agad has been performed to identify the various classes of components present in this formulation. Specifically, HPTLC analysis was performed for the determination of polyphenolic components such as flavonoids and water-soluble antioxidants and acid-base strength compounds.

As shown in [Graph 1], HPTLC-diode-array detection analysis represents the presence of almost five ionic components. One major basic component identified at retention time 29.41 min run time and it accounted maximum proportion among all separated components and proved to be effective analyte in Panchashirishadi Agad Churna. Apart from it, other components like PK No. 30 could be either flavonoid or phytoamine.
Graph 1: HPTLC analysis indicating presence of phytoconstituents in Panchashirishadi Agad (Churna)

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However, comparing [Graph 2],[Graph 3],[Graph 4],[Graph 5] chromatograms, no any significant changes in component quantification were observed. Importantly, very few polyphenols/antioxidants were identified in the chromatograph. In addition, their cumulative presence in extract is very less. Presumably, there might be the chances of the presence of either digoxin or digitoxin such as cardiac glycosides in Panchashirishadi Agad Churna. Moreover, the phytosterols and tocopherols could also be present in Panchashirishadi Agad Churna which were detected at very last part of the chromatograph [Graph 6]. As the quantity of polyphenols in Panchashirishadi Agad Churna was low and even they were hardly detected in reverse phase HPTLC analysis, later the class separation of Panchashirishadi Agad Churna was considered, so that it could confirm the exact quantification of polyphenols as well as ionic components. As shown in [Graph 4], 11 polyphenols/antioxidants were detected in this formulation. In the formulation, 48% of polyphenols were detected which might show the synergistic activities.
Graph 2: HPTLC analysis indicating presence of phenols in Panchashirishadi Agad (Churna)

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Graph 3: HPTLC analysis indicating presence of cardiac glycosides in Panchashirishadi Agad (Churna)

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Graph 4: HPTLC analysis indicating presence of phenols in Panchashirishadi Agad (Vati)

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Graph 5: HPTLC analysis indicating presence of cardiac glycosides in Panchashirishadi Agad (Vati)

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Graph 6: HPTLC analysis indicating presence of phytoconstituents in Panchashirishadi Agad (Vati)

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Furthermore, the class separation using the strong cation exchange chromatography (SCX-HPTLC) was performed. The intention of this study was to investigate the presence of acid-base strength compounds. As shown in [Graph 4], 6% basic alkaline components were identified. There are only two nitrogenous compounds at PK 7, RT value 6.53 (3%) and PK 8, RT time value 6.82 min (3%).

Similarly, to depict any changes in Panchashirishadi Agad Vati form, further analysis was performed using the reversed phase HPTLC (RP-HPTLC). Importantly, during this analysis all retention parameters were kept similar as those used in previous sample (Panchashirishadi Agad Churna) analysis.

Similarly, its class separation including the polyphenols and ionic components detection were performed. Moreover, there retention order was also similar. After the analytical study of Panchashirishadi Agad, it is anticipated that these polyphenolic compounds act as an anti-inflammatory, antiallergic, and analgesic agents and reduce the symptoms developed due to Kita Dhamsha (insect bite).


  Conclusion Top


Panchashirishadi Agad is purely a herbal formulation, which is indicated for Kita Dhamsha, mentioned in Shushrut Samhita, Kalpa Sthana. Panchashirishadi Agad was analyzed on physicochemical and phytochemical parameters. In the HPTLC analysis of this formulation, 11 components (polyphenols/antioxidants) and 6 basic alkaline components were identified.

Polyphenolic compounds are anticipated to possess antiallergic action. The major pharmacological actions present in the ingredients of Panchashirishadi Agad such as antiallergic,7 anti-inflammatory,[8] antioxidant,[9] analgesic,[10] cardioprotective,[11] immunomodulatory activity can suggest its probable mode of action and its preventive and curative aspect on Kita Visha.

Recommendations

  • Through this study, analytical specifications of Panchashirishadi Agad have been established. This has played a crucial role to enhance the ancient knowledge regarding Panchashirishadi Agad, and has also filled gap between ancient and modern knowledge regarding this herbal formulation.


  • Panchashirishadi Agad is indicated as one of the potent formulations for insect bite management. To assess the efficacy of this formulation, further experimental studies on in vivo, in vitro, and on basis of clinical grounds are very essential before bringing it to market level.


  • Further more advanced analytical, in vivo, in vitro studies will prove beneficial for proper standardization of this formulation.


  • Limitations

  • Panchashirishadi Agad contains all the Panchangas of Shirish. Each part has its different chemical constitution. Thus, it contains numerous phytoconstituents. In this study, HPTLC of Panchashirishadi Agad was performed. The various constituents present in this formulation have been detected. But these constituents could not be considered as the markers or standards. Thus, further more advanced studies are required for the study of pharmacodynamics of various phytoconstituents.


  • Financial Support and Sponsorship

    Nil.

    Conflicts of Interest

    There are no conflicts of interest.



     
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    Nguyen NY, Chrambach A. Natural pH gradients in buffer mixtures: Formation in the absence of strongly acidic and basic anolyte and catholyte, gradient steepening by sucrose, and stabilization by high buffer concentrations in the electrolyte chambers. Anal Biochem 1977;79:462-9.  Back to cited text no. 6
        
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    Babu NP, Pandikumar P, Ignacimuthu S. Anti-inflammatory activity of Albizia lebbeck Benth., an ethnomedicinal plant, in acute and chronic animal models of inflammation. J Ethnopharmacol 2009;125:356-60.  Back to cited text no. 7
        
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    Cuzzocrea S, Riley DP, Caputi AP, Salvemini D. Antioxidant therapy: A new pharmacological approach in shock, inflammation, and ischemia/reperfusion injury. Pharmacol Rev 2001;53:135-9.  Back to cited text no. 9
        
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    Panchabhai TS, Kulkarni UP, Rege NN. Validation of therapeutic claims of Tinospora cordifolia: A review. Phytother Res 2008;22:425-41.  Back to cited text no. 10
        
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    Maulik SK, Katiyar CK. Terminalia arjuna in cardiovascular diseases: Making the transition from traditional to modern medicine in India. Curr Pharm Biotechnol 2010;11:855-60.  Back to cited text no. 11
        


        Figures

      [Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6]
     
     
        Tables

      [Table 1], [Table 2], [Table 3], [Table 4], [Table 5]



     

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