Comparative Phyto-Pharmacognostic study of Field collected and Pharmacy sample of Arjuna (Terminalia arjuna) Bark

Dongre Sushma; Pande Shishir

doi:http://dx.doi.org/10.21760/jaims.7.2.2

Comparative Phyto-Pharmacognostic study of Field collected and Pharmacy sample of Arjuna (Terminalia arjuna) Bark

Sushma D.^1*, Shishir P.²

DOI: http://dx.doi.org/10.21760/jaims.7.2.2

^1* Dongre Sushma, Assistant Professor, Dept. of Rasashashtra and Bhaishjya Kalpana, Govt. Ayurved College, Osmanabad, Maharashtra, India.

² Pande Shishir, Ph.D. Guide & HOD, Dept. of Rasashashtra and Bhaishjya Kalpana, ASS Ayurved College, Nashik, Maharashtra, India.

Arjuna (Terminalia arjuna) is the most widely used drug in Ayurveda due to its potential therapeutic competence. Quality assurance is important issue nowadays due to poor collection and storage practices. So, it needs quality control both for small scale and large-scale preparations of Arjuna formulations. Hence present study is planned to compare both field collected and pharmacy sample of Arjuna bark by analytical testing. Macroscopic, microscopic, physicochemical and high performance thin layer chromatography (HPTLC) by using Arjunic acid as a marker is carried out. Macroscopic and microscopic characteristics observed as per Ayurveda Pharmacopeia (API) except color of field collected bark. In Physicochemical parameters, Ash value was higher in pharmacy sample but within limit whereas percentage of water soluble and alcohol-soluble extractives was high in field collected Arjuna as compare to pharmacy sample. Percentage of extractive values was not as per limits of API. Tannin percentage was three times more in field collected Arjuna bark. In HPTLC study, percentage of Arjunic acid was much higher in pharmacy sample (0.072%) than field collected sample (0.054%). Also, the spots observed were more in this sample. 0.60 was the Rf Value for Arjunic acid and one common unknown spot was noticed in both samples. Present study set preliminary data for percentage of Tannins and quantification of Arjunic acid in Arjuna Bark by HPTLC study which is not found in monograph of Arjuna hence can be used as reference for further study.

Keywords: Arjuna, Terminalia arjuna, Pharmacognostic study, Ayurveda, Physicochemical parameters

Corresponding Author	How to Cite this Article	To Browse
Dongre Sushma, Assistant Professor, Dept. of Rasashashtra and Bhaishjya Kalpana, Govt. Ayurved College, Osmanabad, Maharashtra, India. Email:	Dongre Sushma, Pande Shishir, Comparative Phyto-Pharmacognostic study of Field collected and Pharmacy sample of Arjuna (Terminalia arjuna) Bark. J Ayu Int Med Sci. 2022;7(2):7-15. Available From https://jaims.in/jaims/article/view/1663

Introduction

Arjuna (Terminalia arjuna) is well-known drug in Ayurveda which is widely used for many therapeutic purposes. It consists of the stem bark of Terminalia arjuna W. & A. (Fam. Combretaceae); a large deciduous tree, commonly found throughout the greater parts of the country.^[1]

It is widely used for therapeutic purposes in Ayurveda in various forms such as Kshirpaka, Siddha Gruta, Arishta etc. It is indicated in Medoroga (Obesity), Vrana (Wounds), Hridroga (heart diseases), Kshatashaya (Debility), Prameha (Diabetes), Vyanga (Chloasma) etc.

In Ayurveda, bark is collected under good collection practice, when the potency of drug is optimum at that particular season.^[2]

Although bark is widely available in market, it is hardly self-collected for pharmaceutical purposes. The market for herbal drugs has grown at an impressive rate due to a global resurgence in traditional and alternative healthcare systems, and therefore medicinal plants have great economic importance.

However, loss of biodiversity, over-exploitation and unscientific use of medicinal plants, industrialization, bio piracy, together with lack of regulation and infrastructure are the major impediments to the growth of herbal medicine.

In most countries, herbal products are launched into the market without proper scientific evaluation, and without any mandatory safety and toxicological studies. There is no effective machinery to regulate manufacturing practices and quality standards.^[3]

However, Standards of raw drugs in Pharmaceutical companies are maintained to some extend with in-house quality control laboratory. Also, storage condition may impact the quality specially before being supplied to pharmacy. The post- harvesting process of medicinal plants has the great importance in the production chain, because of its direct influence on quality and quantity of active principles in the product sold.^[4]

Due to large scale demand there need to assure quality of Pharmacy sample. Also, self- collected samples need quality check by different analytical testing. So, in present study, comparative study has been planned between Pharmacy

and Self collected sample of Arjuna bark to ensure quality by pharmacognostic evaluation.

Methods and Materials

Collection and Authentication of drug: Arjuna is collected from Forest region of Bhimashanker, near Pune which is authenticated from Botanical survey of India. Whereas pharmacy sample of Arjuna is taken from GMP certified Ayurvedic pharmacy that was authenticated from in-house Quality control department.

Phyto-Pharmacognostic study: Macroscopic, Microscopic study, Physicochemical study and High performance thin layer chromatography (HPTLC) was carried out at Vasu Research Laboratory, Vadodara, Gujrat

Macroscopic and Microscopic study

Macroscopic Study [5]: Macroscopic study was carried out as per standard operating procedures in mentioned in pharmacognosy. Various Morphological characters and organoleptic characterization such as size, shape, color, odor, taste, external markings, fracture, were observed. The diameter of bark was measured with the help of vernier caliper.

Microscopy study [6]: Microscopic identification was carried out according to the Ayurveda pharmacopeia of India (Appendix - 2.1.1)

Physicochemical analysis: Physicochemical analysis namely pH value^[7], Foreign matter^[8], Ash Value^[9], Acid insoluble ash^[10], Water soluble extractives^[11],and Alcohol soluble extractives^[12], were carried out according to the Ayurvedic Pharmacopeia of India (API).

Tannin [13]: Tannin is detected by treating with ferric ammonium sulphate solution (blue-black or green black color shows the presence of Tannin) or with potassium-bichromate solution (brown color indicates the presence of Tannin).

HPTLC analysis of Arjuna (Identification and Quantification)^[14]

Preparation of Test solution: 1 g of Arjuna sample was accurately weighed in a conical flask. It was refluxed for 30 minutes with 15 mL of Methanol consecutively for 3 times. Then filtered with the help of Whatman filter paper No. 1 and concentrated the combined extracts in an evaporating dish on water bath. Thereafter,

made up the volume up to 10 mL with Methanol. Test solution thus obtained was used for HPTLC fingerprinting.

Preparation of Standard solution: Accurately weighed 11.7 mg of standard Arjunic acid was taken into 2 mL volumetric flask and dissolved in Methanol to make up the volume up to 2 mL with Methanol. From this stock solution, 0.1 mL of solution was pipetted into 2 mL volumetric flask and made up the volume up to 2 mL with Methanol. The Standard solution thus obtained was used for HPTLC fingerprinting.

Observations

Table 1: HPTLC Chromatographic Conditions

Chromatographic Conditions
Application Mode	CAMAG Linomat 5 - Applicator
Filtering System	Whatman filter paper No. 1
Stationary Phase	MERCK - TLC / HPTLC Silica gel 60 F254 on Aluminum sheets
Application (Y axis) Start Position	10 mm
Development End Position	80 mm from plate base
Band Length	8 mm
Distance Between Tracks	15 mm
Sample Application Volume	12.0 μL
Standard Application Volume	12.0 μL
Development Mode	CAMAG TLC Twin Trough Chamber
Chamber Saturation Time	30 minutes
Mobile Phase (MP)	Chloroform : Methanol (9 : 1 v/v )
Visualization	@ 254 nm
Quantification Wavelength	205

1) Images of Field Collected T. Arjuna (Image I-IV, VII)

I. T. Arjuna tree

II. Wet bark of T. Arjuna

III. Dry bark of T. Arjuna

IV. Powder of T. Arjuna

2) Images of Pharmacy Collected sample of T. Arjuna (V-VI,VIII)

V. Bark of T. Arjuna

VI. Powder of T. Arjuna

VII. Bark of T. Arjuna (Field collected)

VIII. Bark of T. Arjuna (Pharmacy sample)

Table 2: Macroscopic study of T. Arjuna Bark

Sample Parameters	Macroscopy of Arjuna Field Collected Sample (FCS)	Macroscopy of Arjuna Pharmacy Sample (PCS)
Stem Bark	Stem bark pieces flat or slightly curved	Stem bark pieces flat and slightly curved
Outer surface	Smooth and greenish brown in colour;	Smooth and light brown in colour
Inner surface	Brown and longitudinally striated;	Brown and longitudinally striated;

Microscopy study of T. Arjuna Bark

Pharmacognostic Features of Powder

1. Compound starch crystal