Radical Scavenging Activity Measurement and IC50 Calculation DPPH Assay in Excel

This video explains about the Radical Scavenging Activity Measurement and IC50 Calculation from DPPH Assay in excel

DPPH Assay: Radical Scavenging Activity Assay - Principle, Procedure, Advantages and Limitations

DPPH Assay:
DPPH assay was discovered by Goldschmidt and Renn in the 1920s. DPPH stands for 2, 2-diphenyl-1-picrylhydrazyl. It is a stable free radical with purple colour which turn yellow when scavenged. DPPH is used for measuring the Total Antioxidant Capacity (TAC).

Antioxidant capacity is an overall ability of organisms or food to catch free radicals and prevent their harmful effect.

The % radical scavenging activity of the sample can be calculated using the following formula,

%𝑹𝑺𝑨=(𝑨𝒃𝒔 𝒐𝒇 𝒄𝒐𝒏𝒕𝒓𝒐𝒍 −𝑨𝒃𝒔 𝒐𝒇 𝒔𝒂𝒎𝒑𝒍𝒆)/(𝑨𝒃𝒔 𝒐𝒇 𝒄𝒐𝒏𝒕𝒓𝒐𝒍)∗ 𝟏𝟎𝟎

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[mellicb]

The DPPH assays doesnt show linear results as far as I know. Therefore you can't use linear regression to calculate the IC50. Also, it makes no sense to have an IC50 for each concentration. What you want is to have one IC50 value for one substance, an plant extract for example. The IC50 shows you the amount of extract needed to process the DPPH by 50%. If the highest concentration shows an inhibition ratio/Radical scavenging activity e.g. of 80% (meaning the saturation is at 80%!). you want the calculate the concentration of the extract that has an RSA of 40%. and this is your IC50. The higher your IC50, the more extract is needed to process the DPPH and therefore the lower ist the antioxidative capacity. and all this isn't possible with what you've shown here. I may be incorrect, but this is how I've uderstood it.

[adebayobale]

Thanks a lot for this Video. The IC50 means the value of "y" is 50.

[jeyamanikandanvenkatachalam]

Y represents %RSA; not the concentration.
X represents the concentration of extract.

[TheSingtangpaScienceGuy]

Hi, shouldn't there be only one IC50 value? Your calculation of IC50 for each concentration really confuses me, pliz clarify. thnks

[생약학실험-sv]

Sir, y=9.2788x+28.242 formula was generated when x axis was 1, 2, 3, 4, 5, 6, 7. If the x axis is the concentration, it will be different. Please check it.

[trendingN]

Sir what, if control absorvance reading is lower than the sample absorvance values.

[sarathjohncena]

sir how can we draw the %RSA of control along with the test?

[mohamedrizwan]

Ic50 in your table indicated ug/ml or micromolar concentration ?

[amruthac]

The same calculation Y intercept value is showing different values for us. Please confirm the calculations
or we are getting wrong value means please let us know

[modoumangan]

Thank you sir: How did you get the m slope and b intercept?

[robinakader]

Hi Sir! Thank you so much for the clear explanation about the assay. Can you please tell me how to change the values in x-axis??

[Abiha-ASM]

Ic50 ki samajh nahi aaei as according to u 50ug concentration is more potent ???

[mahfuzalam]

How to calculate adsorption efficiency from UV Vis results ?

[KeepingUpWithIfunanya]

Good afternoon, I really appreciate the video but I am finding it very hard to follow as it is not detailed enough. Especially the parts where you are plotting the standard curve.

[ABCDEFGHIJKLMNOPQ]

Sir how to find the Standard deviation/ error for IC50 plz explain

[christiankeithgiganto]

Sir what does it mean when the ic50 is around 20 to 30?

[zeo]

very helpful! how can we calculate TEAC ?

[digvijaysinghguleria]

Can you make a video on ABTS procedure and calculations??

[smudge]

For Phosphomolybdate assay and FRAP assay we should calculate IC50 ?

[welelameka]

how the concentration of sample increase with decrease the absorbance?