1.0 INTRODUCTION:
Inflammation is a body response to injury,
infection, or destruction characterized by heat, redness, pain, swelling, and disturbed
physiological functions, caused by physical trauma, noxious chemical, or
microbial agents. It is the body's response to inactivate or destroy the invading
organisms, to remove the irritants, and set the stage for tissue repair. It is
triggered by the release of chemical mediators from injured tissue and
migrating cells. The migration of leukocytes from the venous systems to the
site of damage, and the release of cytokines, are known to play a crucial role
in the inflammatory response. These chemicals cause the widening of blood
capillaries (vasodilation) and the permeability of the capillaries. This will
lead to increased blood flow to the injured site.
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2.0 METHODOLOGY:
·
Preparation Of Reference Drug:
· Non-steroidal
anti-inflammatory drugs (NSAID) ibuprofen and steroid prednisolone will be used
as reference drugs.
·
20.0 ml of distilled water will
be added to 0.2 g of fine crushed prednisolone powder and vortex.
·
Similarly, the ibuprofen will
be prepared.
·
Serial dilutions:
· Serial dilution from 1000 µg/ml
to 0.01 µg/ml of the test sample and standard drug will be prepared.
·
All test samples and drugs will
be prepared in 5.0 ml of total volume.
· Reaction mixtures will be
prepared using 2.8 ml of phosphate‑buffered saline (pH 6.4) and 0.2 ml of egg
albumin (from fresh hen’s egg).
· 2ml of the test sample and standard
drug (Positive control) from each different concentration will be mixed gently
with reaction mixtures.
·
Distilled water will be used
as the negative control.
·
Inhibition of protein denaturation
·
Reaction mixtures will be
incubated in a water bath at 37°C ± 2°C for 15–20 min, and later, they will be
heated at 70°C for 5 min.
·
The reaction mixture will be
allowed to cool down at room temperature for 15 min.
·
Absorbance will be measured before
and after denaturation of reaction mixture for each concentration (1000 µg/ml,
100 µg/ml, 10 µg/ml, 1 µg/ml, 0.1 µg/ml and 0.01 µg/ml) at 680nm using a
colorimeter.
·
Each test will be repeated
thrice and the mean absorbance will be recorded.
·
The percentage of inhibition
of protein will be determined on a percentage basis with respect to control
using the following formula.
(%)
Percentage inhibition = Ac – At/Ac × 100
Where
At is the absorbance of the test and Ac is the absorbance
of the control.
3.0 ENDPOINT PARAMETER(S):
·
% Percentage inhibition activity
4.0 REFERENCE(S):
4.1 Dharmadeva S, Galgamuwa LS, Prasadinie C,
Kumarasinghe N. In vitro anti-inflammatory activity of Ficus racemosa L. bark
using albumin denaturation method. AYU 2018;39:239-42.
4.2 Stephanie Flore Djuichou Nguemnang, Eric Gonzal
Tsafack, Marius Mbiantcha, Ateufack Gilbert, Albert Donatien Atsamo, William
Yousseu Nana, Vanessa Matah Marthe Mba, and Carine Flore Adjouzem. In
VitroAnti-Inflammatory and In VivoAntiarthritic Activities of Aqueous and
Ethanolic Extracts of Dissotis thollonii Cogn. (Melastomataceae) in Rats.
Evidence-Based Complementary and Alternative Medicine Volume 2019, Article ID
3612481, 17 pages https://doi.org/10.1155/2019/3612481.
4.3 Shunmugaperumal, T., Kaur, V. In
Vitro Anti-inflammatory and Antimicrobial Activities of Azithromycin After
Loaded in Chitosan- and Tween 20-Based Oil-in-Water Macroemulsion for Acne
Management. AAPS PharmSciTech 17, 700–709 (2016). https://doi.org/10.1208/s12249-015-0401-2.
END OF DOCUMENTS
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