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How we prepared sample plant extracts?

We were keen to examine pharmacognostic and pharmacological activities of wild orchid species of Nepal. Firstly, we seek permission from authorities to collect  orchid species (total 11 species) in a limited amount (from 250 - 500 g of fresh material according to availability at collection site) for the research purpose. After getting a letter of permission, we went to community forests of Mid-western, and Central Nepal to collect them. The sample specimens were authenticated, dried, extracted and tested for the pharmacognostic and pharmacological activities. Here I am presenting briefly about the way of collection, drying, and extraction.   

1. Collection of Plant Material
A pseudobulb of Otochilus ablus with leaf, flower, and fruits 

We collected mature and healthy plant from a place where the population of selected plant species was relatively higher . Then, a specimen of selected orchid  with flower was presented to Dr. Bijaya Pant (Associate Professor, Central Department of Botany, Tribhuvan University, Nepal) for the authentication and during field trip to community forest Mr. Bhakta Bahadur Raskoti (Scientific officer, Plant Scientist, Nepal Academy of Science and Technology, Nepal) also identified and authenticated.


 2. Drying of Plant Materials
We washed plant materials (root, pseudobulbs, stem, and leaf) under tape water; kept them under shade to dry up remaining water content. We noted the mass of fresh plant material which was from 250 - 500 g. We chopped plant materials into small pieces by commercially available knives and left a mass of small pieces under the shade for two weeks to dry. We measured the  mass of dried pieces on a digital balance and continued drying and weighing till it gave constant reading. Then, we noted final reading of mass of dried pieces which was one tenth of the fresh material. Finally, we made dried pieces of plant material into fine powder by commercially available electric blender.
3. Extraction of Plant Powder

Rotary evaporato
We applied the hot continuous Soxhlet extraction technique to get concentrated plant extract. For this, wetook the fine powder of plant material (20 g) in thimble and poured ethanol ( 70 %; 200 ml) into boiling flask. The Soxhlet extractor was fitted to work properly and continuously run for 4 hours. We dried concentrated plant extract in a rotary evaporator to get dark brown coloured paste. The dark brown coloured paste was kept undisturbed in a petridish under shade to dry up remaining portion of solvent. Then, we collected dark brown powder (which was found to be one fifth to one tenth of dry powdered material) in tiny glass jar and kept in a refrigerator (temperature 4 degree Celsius) for further experiments.

 

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