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: Radical Sc...
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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[@mellicb9681]

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.

[@shownist]

So, How to calculate IC50 from antioxidant activities extract? I read only using the linear regression :(

[@Dr.Marium_Begum]

you are right

[@emelakbaba1975]

yes, I agree. How come the more concentrated extract has the highest IC50 value? So, this calculation is not correct. one extract must have one IC50 value.

[@DrAkhtarAli08]

I did with formula, IC50 values shoed 1679 which is looking totally rubbish.

[@titiyahomie]

Hello. Please if a linear regression can’t be used. What statistical tool is then used to calculate IC 50

[@adebayobale6594]

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

[@TheSingtangpaScienceGuy]

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

[@aminatehari6855]

EXACTLY !

[@ChristianaOkorieuku]

Thank you so much, the video was very helpful

[@user-sn4hb4oz2v]

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.

[@jeyamanikandanvenkatachalam]

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

[@robinakader1841]

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??

[@BioResource]

Thank you. Consider subscribing.

[@elyaskhan5212]

Can u please us how to make this formula

[@trendingN608]

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

[@amruthac7633]

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

[@mohamedrizwan7959]

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

[@nagapriyasathishkumarp3748]

Thank you so much

[@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.

[@sarathjohncena3540]

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

[@modoumangan9851]

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

[@MsPooja1980]

very helpful video....

[@BioResource]

Thanks. Consider subscribing 😊

[@mahfuzalam1975]

How to calculate adsorption efficiency from UV Vis results ?

[@christiankeithgiganto5097]

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

[@sahibraja3059]

I think there is no sense of calculating IC50 Value of each concentration.

[@Learningnfun643]

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

[@zeo2813]

very helpful! how can we calculate TEAC ?

[@smudge321]

For Phosphomolybdate assay and FRAP assay we should calculate IC50 ?

[@ABCDEFGHIJKLMNOPQ209]

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

[@digvijaysinghguleria8556]

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

[@user-qj1oq4jy9w]

I would love to get it too

[@shailindkhar1538]

How do you got the m and c value in the equation y=MX+C sir????

[@welelameka]

how the concentration of sample increase with decrease the absorbance?

[@muhammadyahya837]

Who we anlaz for corelation with TFC

[@prafull07prajapati83]

Hello Sir you Share a very good knowledge, but looking video is so bad, plz upload again with clear visible

[@BioResource]

Hi, Please change your video resolution to 720p it will be clear.

[@aayushikoshariya7994]

Sir what is absorbance of control , and what is control

[@luisormenotasayco2287]

Control is just DPPH with a bit more of methanolic sol., it is supposed to be that 520nm is the top of absorbance as it is the higher absorbance researchers use the most

[@deepakkhatak9271]

Yes you are sharing wrong information regarding IC50 Value please correct it

[@surjitdeka7092]

What should be the r square value..?

[@niluk1827]

My plants treatments is 13 how I put 13 concentration digit

[@BioResource]

Hi, Your question is not clear. Could you elaborate or share the data?

[@firryman5883]

bro please how to analysed dpph data by prism software

[@nasytadifafirdaus3477]

The absorbance is must 0.2-0.8, isn't?

[@chandnipatel9236]

Can u explain how to calculate p value

[@BioResource]

Hi Chandni Patel, Please see the below link for video on How to calculate p value: kzbin.info/www/bejne/a16rqaBqpJZ7iqM Hope this helps.

[@chandnipatel9236]

@@BioResourcep value for antioxidant activity.which value should I take for calculating p value wether absorbance or percentage of inhibition

[@khifsarajpoot707]

As this as vedio again .speak loudely and clear the screen .date presented here showed blured

[@BioResource]

Hi, Please set your video resolution to 720p or 1080p to get clear video.

[@D-bright237]

Nice video but very blurry

[@BioResource]

Hi...please adjust the resolution to 720p or 1080p to get good clarity.

[@biologicalmints9510]

where is standard graph

[@hindutraditionalmedicineht8776]

IC50 is the concentration of extract at which 50% radical is scavenge. The obtained IC50 value is lower than you taken the concentration..... Wrong Concept....