Macro-microscopic evaluation, physicochemical analysis, standardization and HPTLC finger printing of aril part of Myristica fragrans Houtt.

1. Introduction

Myristica fragrans Houtt. is a well known medicinal plant known as Jayaphala in Hindi¹. It is an evergreen moderate sized aromatic tree with greyish black bark having lenticular spots on outer side and red juice on inner side². The plant is native of Moluccas now cultivated in many tropical countries of both hemispheres. In India, it is grown in Nilgiri, Coimbatore, S alem, Ramanathapuram, Tirunelveli, Kanyakumari etc³ . In Sharangadharasamhita Purvakhanda, Deepana pachanadi adhyaya, Jatiphala is mentioned as Shukra sthmabaka dravya⁴ and also in the madhyama khanda, churna Kalpana adhyaya it is mentioned that the Jatiphaladi churna is beneficial in kasa, shwasa, aruchi, kshaya etc. conditions.

In Atisara chikitsa of Bhavapraka shasamhita, uttarardha, Jatiphala is mentioned as one of the dravya of Vijayava leha⁵ .

M.fragrans has been reported with the chief chemical constituent Myristicin⁶. other than that, some of other phytochemical constituents present in different parts of Myristica fragrans are; Alpha-terpinine, Alpha-thujene, Amylodextrin, caprylicacid, Malabaricane B, Malabaricane C, Calcium, carbohydrates etc.

And also the kernels of Myristica malbarica Lam. are sometimes mixed with the material. Also the fruits of Myristica dactyloides Gaerth are mixed with the fruits of Myristica malabarica Lam for the purpose of adulteration ⁷,⁸ .With this background, detailed quality control studies were undertaken for this traditional raw drug with the aim of developing standards of authenticity.

2. Materials and Methods:

2.1 Collection and Identification

Dried aril part of Jatiphala were collected from local Ayurvedic shop in Udupi, Karnataka. The plant material was authenticated at Pharmacognosy department of SDM Centre for Research in Ayurveda and Allied Sciences, Udupi, Kuthpady, Karnataka. And (a specimen code 16032903) is being mentioned for further reference. The dried aril portions of Jatiphala were cleaned, coarsely powdered and used for macroscopic and microscopical characterization, phytochemical analysis and HPTLC.

2.2 Macro-microscopic analysis:

The external features of the test samples were documented using Canon IXUS digital camera. The macroscopic features were compared to local flora for authentication.

Microscopy

Sample was preserved in fixative solution. The fixative used was FAA (Formalin-5ml + Acetic acid-5ml + 70% Ethyl alcohol-90ml). The materials were left in FAA for more than 48 hours. The preserved specimens were cut into thin transverse section using a sharp blade and the sections were stained with saffranine. The slides were also stained with iodine in potassium iodide for detection of starch. Transverse sections were photographed using Zeiss AXIO trinocular microscope attached with Zeiss Axio Cam camera under bright field light. Magnifications of the figures are indicated by the scale-bars.

Powder microscopy

A pinch of powder was warmed with drops of chloral hydrate on a microscopic slide and mounted in glycerine. Slides observed under microscope and diagnostic characters were observed and photographed using Zeiss AXIO trinocular microscope attached with Zeiss AxioCam camera under bright field light. Magnifications of the figures are indicated by the scale-bars

2.3 Physico chemical analysis:

The results of the Physico chemical Analysis are mentioned in the below mentioned Table-1.

2.4. HPTLC finger printing

1gm of Jatiphala powder was extracted with 20 ml of alcohol, kept overnight and filtered . 4, 8 and 12µl of the above extract was applied on a pre-coated silica gel F254 on aluminum plates to a band width of 7 mm using Linomat 5 TLC applicator. The plate was developed in nhexane: Chloroform (1.0: 1.0). The developed plates were visualized in UV 254, 366, and then derivatised with anisaldehyde sulphuric acid reagent and scanned under UV 254nm, 366nm and 620nm. R, colour of the spots f and densitometric scan were recorded.

1. Results:

The results of macro and microscopy Standardization parameters and HPTLC (Photo documentation densitometric scan of Rf values) are presented in above mentioned respective tables and figures.)

2. Discussion:

The seed portion and aril portion of Myristica fragrans are well known Shukra sthambaka and Atisaraghna dravya in Ayurveda System of medicine. Morphological and anatomical standardization of herbal drug needs the information from basic disciplines of plant sciences for identification of plant drug.

Simultaneously for identification of chemical nature of plant in term of physico-chemical analysis, qualitative and quantitative analysis for the detection of active constituents, expertise are required. According to Kunle et.al, Standardization of herbal drug is aseries of Protocol which assure the quality, efficiency and safety of plant drug ⁹. Macro-microscopic analysis helps in the identification of plant characters anatomically and helps in identification of botanical background. Standardization and authentication of plants was done by evaluating physico chemical testing¹⁰. The values obtained in the study will serve as constants for quality standard measures for standardization of drugs in the dried form.

High performance thin layer chromatography (HPTLC)serves as the quality assessment tool which helps in identification of variation in chemical composition of plants. TLC identity is a part of every herbal monograph of international standards.¹¹ HPTLC fingerprinting shows different Rf values at different wave lengths and reported values can be used as a quality indicating fingerprint for Myristica fragrans in the dried form. Findings reported in the present investigations are in support of API and QSIMP2012.¹²

3. Conclusion:

Pharmocognostical characterization of the Myristica fragrans has been done as per pharmacopeial methodology. Present study explores the botanical (in terms of macro microscopic observations) physicochemical observations(in terms of total ash, AIA, ASA,ASE,USE and loss on drying ) and HPTLC fingerprint profile can serve as excellent standard for the identification and authentication of drug in dried form.

Conflict of interest:

All authors have none to declare.

Acknowledgements:

The author is highly grateful to Dr. Ravirao Sorake; Professor, Department of Dravyaguna; Alva's Ayurvedic Medical College and Hospital; Moodabidri. Author highly regard Dr. B Ravishankar, Director, SDM Centre for Research in Ayurveda for providing lab facilities. Dr. Sunil Kumar and Dr. Suchitra N. Prabhu are gratefully acknowledged for constant support during the study.