Experiment No.

02
Date: 2077/9/10
PREPARATION OF PLANT EXTRACTS FOR PHYTOCHEMICAL ANALYSIS

1.0 Introduction
1.1 Phytochemical

Phytochemicals, which are non-nutritional components of plant origin, are one of

the nutritional factors that play significant role in various functions of the animal body (Lee et
al., 2017). Many of these compounds found in food products are known to have antioxidant
potential due to the occurrence of the OH group (Mahmood et al., 2019). Phytochemical also act
as protective action toward plant; thus, it protects plants from disease and damage and affects the
color, aroma, and taste of the plant. Typically, plant chemicals that protect plant cells from
harmful environmental factors such as pollution, stress, drought, ultraviolet radiation, and
exposure to pathogenic microorganisms are called phytochemicals. Recently, it is known that
they have roles in the protection of human health when their dietary intake is significant. More
than 4,000 phytochemicals have been cataloged and are classified by protective function,
physical characteristics, and chemical characteristics (Velavan, 2015). These phytochemicals,
alone and/or in combination, have enormous therapeutic potential to cure various diseases (Prof
et al., 2012).

1.2 Neem

Neem plant (Azadirachta indica L.) is considered one of the most important medicinal plants in
the world. The medicinal properties of this plant are described in the ancient documents "Charak-
Samhita" and "Susruta-Samhita", which are believed to be the basis of Ayurveda, the natural
healing system of India (Girish & Shankara, 2008). It is considered "Sarvaroganivarini"
which means the healer of all illnesses (Subapriya & Nagini, 2005). Different parts of the neem
plant are used to treat many diseases of the Ayurvedic medical system around the world. The
aqueous extract of neem bark has therapeutic potential for controlling gastric hypersecretion and
gastroduodenal ulcer (Bandyopadhyay et al., 2004), while neem leaf extract has been used to
reduce oral infections, plaque index and bacterial count (Pai et al., 2004).
2.0 Materials & methods

2.1 Plant material preparation

Fresh leaves from of neem was collected from Itahari, Sunsari, Nepal. The entire plant was
washed and dried at 50℃ in a tray dryer. After drying, the plant materials were powdered well
by using grinder.

2.2 Preparation of plant extracts for phytochemicals analysis

The phytochemical extraction was performed using organic solvent extraction as well as aqueous
extraction.

The organic extraction was performed by Soxhlet extraction method. the extraction was done by
taking 20 gm of dried plant powder and was placed into a glass thimble then extracted with 250
ml of different solvent separately (ethanol and methanol). The extraction process was carried on
till the solvent in the siphon tube of Soxhlet apparatus become colorless. After that the extract
was heated on hot water bath at 35℃ until all the solvent evaporated. The dried plant crude
extract was kept in refrigerator at 2-8℃ for their future use.

The aqueous extraction was done by taking 5 gm of the plant powder and mixed with 200 ml of
distilled water in a beaker. The mixture was heated on a hot plate at 30-40℃ and mixed
continuous stirring for 20 minutes. The mixture was filtered using Whatman filter paper and the
filtrate was used for the further phytochemical analysis.

3.0 Results and Discussion

Hence, the plant extracts were prepared for phytochemical analysis.

4.0 References

Bandyopadhyay, U., Biswas, K., Sengupta, A., Moitra, P., Dutta, P., Sarkar, D., Debnath, P.,
Ganguly, C. K., & Banerjee, R. K. (2004). Clinical studies on the effect of Neem
(Azadirachta indica) bark extract on gastric secretion and gastroduodenal ulcer. Life
sciences, 75(24), 2867-2878. https://doi.org/10.1016/j.lfs.2004.04.050
Girish, K., & Shankara, B. S. (2008). Neem–a green treasure. Electronic journal of Biology, 4(3),
102-111.

Lee, M. T., Lin, W. C., Yu, B., & Lee, T. T. (2017). Antioxidant capacity of phytochemicals and
their potential effects on oxidative status in animals -a review. Asian-Australas. J. Anim.
Sci., 30(3), 299-308. https://doi.org/10.5713/ajas.16.0438

Mahmood, M. H., Osama, A. K., Makky, E. A., Rahim, M. H., Ali, N. H. M., & Hazrudin, N. D.
(2019). Phytochemical Screening, antimicrobial and antioxidant efficacy of some plant
extracts and their mixtures. IOP Conference Series: Earth and Environmental Science,
346. https://doi.org/10.1088/1755-1315/346/1/012003

Pai, M., Acharya, L. D., & Udupa, N. (2004). The effect of two different dental gels and a
mouthwash on plaque and gingival scores: a six‐week clinical study. International dental
journal, 54(4), 219-223. https://doi.org/10.1111/j.1875-595X.2004.tb00283.x

Prof, D., Gupta, C., & Sharma, G. (2012). Importance of Phytochemicals in Nutraceuticals. J.
Chinese Med. Res. Development, 1, 70-78.

Subapriya, R., & Nagini, S. (2005). Medicinal properties of neem leaves: a review. Current
Medicinal Chemistry-Anti-Cancer Agents, 5(2), 149-156.
https://doi.org/10.2174/1568011053174828

Velavan, S. (2015). Phytochemical techniques - a review. World J. Sci. Res., 1(2), 80-91.