I am performing in vitro anti-inflammatory assay: inhibiton of albumin denaturation by organic extract and purified fractions from natural material. Since my samples are colored (red, orange, yellow), I keep getting negative vaules, thus indicating pro-inflammatory rather than anti-inflammatory effect of extract/fractions. Concentration range of samples is 1 mg/ml to 62 ug/ml and the turbidity (absorbance at 660 nm) is higher with the increase of concentration. The results are the same even when I subtract the absorbance of blank (pure extract/fractions which almost do not absorb at wavelength mentioned above).
I am calculating the inhibition according to formula:
% Denaturation inhibition = (1−D/C)×100%
Where D is the absorbance reading of the test sample, and C is the absorbance reading without test sample (negative control).
I am measurng turbidity in 96-well plate; the solvent is methanol.
I was wondering if I am doing something wrong with my experiment setup or do you have any ideas how to avoid this problem?
According to the literature, there should be compounds in my extract/fractions that have already showed anti-inflammtory effects.
I only show our data as for OVA Adsorption Bio Assay by using "Inorganic Particles". In this system, albumin alternation were not observed and Inflammatory materials suppression data were very clearly shown.
I am very doubtful for using organic substance for reducing inflammatory materials because interaction between test samples may be caused and also
solvent's damages.
- Badges
- Science topic