I have always loved your detailed explanations of the concerned assays. Here, is there any formula to calculate the % inhibition like that in DPPH. Please provide if there is any. Also please make a detailed video on protocol of H2O2 scavenging activity, and enzyme inhibitory assays.
I love the way u explain Sir, I can understand clearly.. thank you very much sir.. there no much FRAP experiment I found in youtube. Ur video helps alot
Hi , Whatever you are presenting is Potassium ferric cyanide reducing antioxidant power(PFRAP) but not FRAP where one should use TPTZ. Also note that one must use potassium Ferric cyanide not potassium ferrocyanide.
Thankyou so much Bro, for sharing this, plzzzzz make video on ABTS antioxidant assay, and lets make it easier for us to understand, it my viva on end of April, and your videos are helping me much, plz make video on ABTS ASAP!!!!!! PLZ
Your video is very helpful for all scholars and students. but you have not shown data interpretation for FRAP assay. I have some doubt to relate this assay. please can explain about it.
Hello sir Video is very informative and useful. Sir I have seen potassium ferricyanide instead of ferrocyanide used in this video in other research journals. Please clarify the reagent used potassium ferricyanide or ferrocyanide.
I have published all procedures in my paper they all are modified by me. The details are given in a separate video in my channel pls watch that and cite that paper
Hello thank you for the video it was helpful. Although the solution turned green not blue i wonder why (my ferricyanide solution was yellow). And by the way can we store the remaining of the solutions for an ulterior use ? (Ferry chloride and ferricyanide solutions)
Thank you for your clear explanation... I guess it's 1% Potassium Ferricyanide.... I hv seen a different protocol for FRAP using TPTZ and what u hv shown here is reducing power assay right?
Sir hope u will b f9 n thanks 4 ur response to our queries... Sir plz tell me I should plata graph between absorbance n concentration directly or I should plata graph between percent inhibition n concentration?
Thank you for efforts and clarity. However I have a question: can we use only one concentration I saw in quite a few Protocol that it makes only one concentration and they report the results in terms of mg equivalent of Ascorbic Acid, is it correct or wrong? like this protocol : " The FRAP activity of the extracts was measured according to the method previously described [11]. Briefly, 5 μL of ethanolic extract was mixed with 900 μL of FRAP reagent and 95 μL of water (FRAP reagent was prepared to mix acetate buffer (300 mM, pH 3.6): TPTZ solution (10 mM in chloride acid) and ferric chloride solution (20 mM) (10:1:1 v/v). Absorbance at 593 nm was measured with an Agilent 8453 UV-Vis spectrophotometer, 4 min after starting the reaction. Results were expressed as ascorbic acid equivalents (AAE) per gram of dry weight (DW). Each determination was performed in triplicate and repeated at least three times"
For positive control what we gonna have? All chemicals minus plant extract? Or All chemicals along with methanol? Do i need to add methanol in the positive control test tubes?
Is there any calculation to display the result of this assay or just just have to plot a graph of different absorbance values in different concentration? Also, why taking negative control in this assay as there is no role of this value in calculation part?
Hello sir can you please explain us Determination of superoxide anion (O2 −), malondialdehyde (MDA), and hydrogen peroxide (H2O2) also Extraction and assay of enzymes APX, CAT, SOD, and POX
Sir when we are doing standard for any antioxidant activity assay.. and suppose we are using ascorbic acid as standard, then we need to disssolve it in same solvent used for extract preparation or disssolve in distilled water? Thank you sir
Is it necessary on the same day when we do experiments of ddph and frap on that Same day we do spectrophotometer analysis is it possible that on one day we do experiments can we do spectrophotometer analysis next day or not plzz so tell?
No there's no formula, whatever I have mentioned in video that's all. You can access my paper through Google Scholar or research Gate and go through the process/results
In this video u said add 2.5 ml of TCA and then 2.5 ml of d. Water. But in the article it's quantity is written as 250 ul of solution and 250 ul of water. So what quantity should i consider? Kindly reply asap.
Sir kindly tell me absorbance of antioxidant is directly related to frap or inversely like DPPH? Because if absorbance increases with increase in concentrations then percent inhibition decreases ... kindly give me guide line
@@Dr.Rakesh.B from which website can I get the recipe? I got a recipe for monobasic heptahydrate, but I have monobasic dihydrate, how can I convert? Can I get a recipe for mono and dibasic dihydrate?
Sir I am working on nano particle green synthesis . I did for selenium metal . After adding dpph the colour still stays so dark and the reading shows error. Should I dilute or check ph like that before antioxidant assay
Sir I am working on nano particle green synthesis using plant extract and doing antioxidant assay for it . But once I add dpph it’s too dark and the reading also says error Can u guide me as to how to go about this. Should I do a dilution or try any other method . The nano particle i synthesised was selenium
Hello sir, Could you please share the reference material and the paper from which you may have received this protocol for? I would like to fully understand what each material's purpose is and what reagents can be used as replacements, especially as the Benzie and Strain paper seemed to use TPTZ instead of potassium ferricyanide and TCA. Please let me know if this can be done. Thank you.
You can mail me to rakeshb96@yahoo.com ill send you the reference. And there are more than 10 methods with different chemicals and principles for every assay. You can stick on to any of the protocol according to the availability of chemicals with you
Please watch a separate video I made in my channel for references. You will find my paper you can download and cite the same. If you did not find mail me I'll send my paper rakeshb96@yahoo.com
@@Dr.Rakesh.B sir, i have one doubt . please clarify that. I have chosen my drug with conc.5,50,100,250,500 ug /ml for FRAP in that 5 ug conc. Itself has more colour which is not readble by UV spec. So that i have planned to reduced my drug conc below 5ug /ml .if it so am i reduce the quantity of reagents like instead of 2.5 ml of PBS use 1ml of PBS ...?
Not required. Bcoz only your concentration is decreasing, but your total volume will be same. Because you are going to make it up to same volume. So don't have to reduce it
Hello sir sir plz when u see this MSG plz reply sir m searching for prap assay invitro method it's my project but m not getting it anywhere sir can u please help me out sir plz
I had taken data for different concentrations (100-1000 ug/ml) for my four samples.... And the absorbance is increasing for every concentrations... How will I decide.... Which concentration give best results?
And how I will conclude all this data on my results.... It's very confusing... There's very less video available in frap assay... Help me to clear my doubts🙏
I have always loved your detailed explanations of the concerned assays. Here, is there any formula to calculate the % inhibition like that in DPPH. Please provide if there is any.
Also please make a detailed video on protocol of H2O2 scavenging activity, and enzyme inhibitory assays.
I love the way u explain Sir, I can understand clearly.. thank you very much sir.. there no much FRAP experiment I found in youtube. Ur video helps alot
Thank you. Means a lot :)
my m.phill research is on antioxident activity.. Bro your videos are much usefull.. just keep it up. love from university of lahore
i am very proud of sathasivam and manikkam sir
You have explained nicely Sir. And I kindly ask you to provide alpha-tocopherol assay by HPLC method.
Hi , Whatever you are presenting is Potassium ferric cyanide reducing antioxidant power(PFRAP) but not FRAP where one should use TPTZ. Also note that one must use potassium Ferric cyanide not potassium ferrocyanide.
Maa Shaa Allahh
..Good effort
Thankyou so much Bro, for sharing this, plzzzzz make video on ABTS antioxidant assay, and lets make it easier for us to understand, it my viva on end of April, and your videos are helping me much, plz make video on ABTS ASAP!!!!!! PLZ
You should do ACE inhibitory assay video. Can't find any on youtube. Your videos helped me a lot! Thank you so much!
Sure ok:)
appreciate u from pakistan respectable sir
Many thanks to you for making this video. very useful, clearly explained. well done sir. Hope to see more such videos.
Your welcome :)
Could you please share the mathematical equation by which you calculate the FRAP value. It will be a great help. Thank you.
Thank u so much...please make video on catalase and peroxidase assay also...
Your video is very helpful for all scholars and students. but you have not shown data interpretation for FRAP assay. I have some doubt to relate this assay. please can explain about it.
Wow a clear explanation! Thank You
Hello sir
Video is very informative and useful. Sir I have seen potassium ferricyanide instead of ferrocyanide used in this video in other research journals. Please clarify the reagent used potassium ferricyanide or ferrocyanide.
Both is correct
OK thank you very much sir.
I am research scholar under Calicut university. Could you please sent procedures of analyses through my mail if I sent mail to you.
I have published all procedures in my paper they all are modified by me. The details are given in a separate video in my channel pls watch that and cite that paper
Hello thank you for the video it was helpful. Although the solution turned green not blue i wonder why (my ferricyanide solution was yellow). And by the way can we store the remaining of the solutions for an ulterior use ? (Ferry chloride and ferricyanide solutions)
Thank you for your clear explanation... I guess it's 1% Potassium Ferricyanide....
I hv seen a different protocol for FRAP using TPTZ and what u hv shown here is reducing power assay right?
I have been looking for this comment, do these methods work the same?
Amazing video sir. Could you please do a video on ABTS assay and oxygen radical absorbance capacity??
Thankyou so much for this.., please upload videos for hydrogen peroxide assay
i need it too..
please make a video on in vitro antidiabetic activity
amazing...sir can u make video
s about anyhocyanis and tannins
Nice explanation...🙂....plzz sir make video on ABTS assay...
Sir also perform pancreatic lipase inhibitory assay.
Always love your videos! A Query - So we would be using the units mmol/litre with the absorbance value itself and no further calculation?
There is calculations. Please watch my standard graph video in my channel for further calculations
Good job done. Please, can I get antibiotic from you to enhance my study on plant extract. Thank you
.
I'm sorry, im not working in that lab anymore. You can buy from medical shops it's pretty cheap.
Thanks a lot. What I was trying to request for was antibiotic disc and not antibiotic as stated earlier. I really appreciate your effort. Cheers
Superb video sir. Can you plz send the refrence for the same?
In my channel, I have made a video for all my references please watch it. Or you can follow me in research gate my papers are available for references
Weldone sir... nice job u have done... sir will u plz share a video of ABTS scavenging assay
Ok will try thank you :)
@@Dr.Rakesh.B Thank u sir
Sir hope u will b f9 n thanks 4 ur response to our queries... Sir plz tell me I should plata graph between absorbance n concentration directly or I should plata graph between percent inhibition n concentration?
Same question sir
Percentage inhibition and concentration
@@Dr.Rakesh.B thank u sir... stay blessed
As salaamu alaikum can u upload the video for the calculation part of this assay as soon as possible
hello sir. can you please make videos on Ferric Thiocyanate assay (FTC)and Thiobarbituric assay (TBA).it will be a great help sir.
Thank you for efforts and clarity. However I have a question: can we use only one concentration I saw in quite a few Protocol that it makes only one concentration and they report the results in terms of mg equivalent of Ascorbic Acid, is it correct or wrong?
like this protocol :
" The FRAP activity of the extracts was measured according to the method previously described [11]. Briefly, 5 μL of ethanolic extract was mixed with 900 μL of FRAP reagent and 95 μL of water (FRAP reagent was prepared to mix acetate buffer (300 mM, pH 3.6): TPTZ solution (10 mM in chloride acid) and ferric chloride solution (20 mM) (10:1:1 v/v). Absorbance at 593 nm was measured with an Agilent 8453 UV-Vis spectrophotometer, 4 min after starting the reaction. Results were expressed as ascorbic acid equivalents (AAE) per gram of dry weight (DW). Each determination was performed in triplicate and repeated at least three times"
Can you make video about Evaluation of Anti-inflammatory Activities plz
Ok
For positive control what we gonna have? All chemicals minus plant extract? Or All chemicals along with methanol? Do i need to add methanol in the positive control test tubes?
Yes. Pls watch it again slowly ull understand.
May you kindly tell me how you prepared monobasic and dibasic sodium phosphate
can you tell how the two chemical for the buffer is made how much D/W and chemical is used
Is there any calculation to display the result of this assay or just just have to plot a graph of different absorbance values in different concentration? Also, why taking negative control in this assay as there is no role of this value in calculation part?
No calculations just absorbance
Hello Soumi Paul u have done this assay? Kindly tell me the range of absorbance of ur samples? I mean to say it is above 2 or below 1?
Sir what about the Blank.. it will also develop blue colour?
In meauring absorbtion what will be the base line water or the solvent used for extraction?
Hi sir can u plzz provide the formula which i use end of the experiment and get the value of frap sample after the absorbance
For essential oil can we follow the same protocol or any other protocol for this?
Same
Sir we should check absorbance soon after adding FeCl3 or we can check it after a day or 2?
Soon after adding
Ok thanks
Hello sir can you please explain us Determination of superoxide anion (O2
−), malondialdehyde
(MDA), and hydrogen peroxide (H2O2) also Extraction and assay of enzymes APX, CAT, SOD, and POX
Sir when we are doing standard for any antioxidant activity assay.. and suppose we are using ascorbic acid as standard, then we need to disssolve it in same solvent used for extract preparation or disssolve in distilled water?
Thank you sir
What is the control of this assay? ferric chloride or potassium ferrocynide?kindly guide me....
Control will be all your reagents without plant extract.
Could you plz explain Fenton reaction for anti oxidant activity
Sir, please provide frap assay procedure using TPTZ reagents..........plsssss
I haven't performed that assay. I don't have. Search in Google Scholar
Is it necessary on the same day when we do experiments of ddph and frap on that Same day we do spectrophotometer analysis is it possible that on one day we do experiments can we do spectrophotometer analysis next day or not plzz so tell?
No on same day you have to do
@@Dr.Rakesh.B thank u
Sir big fan of u.
How much quantity should we take into the microplate reader to check absorbance?
200 microlitre i took. Its maximum capacity was 300. It varies from type to type
@@Dr.Rakesh.B okay. Thanks for your reply. These videos are really helpful! God bless you! 💫
Sir is it necessary having buffer of 6.6 ph can it be fluctuate...
It can slightly vary 0.1 to 0.5 max
Sir
Can you please give me the reference of these procedure i need to quote in my research work.
Drop me a mail ill send you papers rakeshb96@yahoo.com
Hlo sir, can you please provide me the paper from where you followed the protocol...it will be a great help sir.
th-cam.com/video/S_hxotaRhwQ/w-d-xo.html
Watch this video and description for the references
Sir can u plz share how to calculate FRAP value ... sir is there any formula to calculate FRAP VALUE?
No there's no formula, whatever I have mentioned in video that's all. You can access my paper through Google Scholar or research Gate and go through the process/results
In this video u said add 2.5 ml of TCA and then 2.5 ml of d. Water. But in the article it's quantity is written as 250 ul of solution and 250 ul of water. So what quantity should i consider? Kindly reply asap.
Both are correct. Use whichever you want. Just that its reduced by 10 times
Sir kindly tell me absorbance of antioxidant is directly related to frap or inversely like DPPH? Because if absorbance increases with increase in concentrations then percent inhibition decreases ... kindly give me guide line
It is Inverse similar to dpph
@@Dr.Rakesh.B thank u sir u will always be In my prayers... u made my research work easy... stay blessed
Sir what is difference between ABTS and DPPH assay i mean to say why more than assay considered in the article
Every method has its own principle so we estimate in as many methods to confirm
Wht compounds can we used to plot standard curve??
Plz reply
Same chemicals except plant extract. Everything is mentioned in video in detail pls watch carefully
@@Dr.Rakesh.B that means feso4+water+frap solution
Which is the cheapest procedure for knowing antioxidant content of a drink ?
You can give to a food safety lab. Thats cheapest and more dependable
@@Dr.Rakesh.B yah done that finally 😊 ur videos are very helpful though. Thank u
Sir kindly share protocol of ABTS assay
I have not performed ABTS assay so I don't have that protocol
Good explanation sir
sir, which reagent give blue colour in this assay?
Sir kindly provide vedios of nitric oxide and hydrogen peroxide assay also.
@@Dr.Rakesh.B sir please do video for Thiobarbituric acid method sir
What if the compound is a powder of pure compound? Not being an extract what ro do then?
Sir plzz tell how to do dilution calculations ..and how to make dilution from stock
Thanku sir
I need a protocol to determine EGCG in tea
Can you explain how to prepare the phosphate buffer?
It's not complicated you have to just add 2 components that's it. You can refer that book I mentioned or website
@@Dr.Rakesh.B from which website can I get the recipe? I got a recipe for monobasic heptahydrate, but I have monobasic dihydrate, how can I convert? Can I get a recipe for mono and dibasic dihydrate?
How to calculate frap using standard graph
Which other reagent can I use, if I don't have trichloroacetic acid (TCA)?
Sorry i haven't tried with any other chemicals to help you in this regard
Thank you for your response
Hello sir , could you pls share the paper for reference. Which protocol are you following?
absorbance taken at UV spectroscopy or IR?
UV
Sir I am working on nano particle green synthesis . I did for selenium metal . After adding dpph the colour still stays so dark and the reading shows error. Should I dilute or check ph like that before antioxidant assay
Sir I am working on nano particle green synthesis using plant extract and doing antioxidant assay for it . But once I add dpph it’s too dark and the reading also says error
Can u guide me as to how to go about this. Should I do a dilution or try any other method . The nano particle i synthesised was selenium
Sir its potassium farrocynide or farricynaide?
Potassium ferrocyanide
@@Dr.Rakesh.B thanks
Hello sir,
Could you please share the reference material and the paper from which you may have received this protocol for? I would like to fully understand what each material's purpose is and what reagents can be used as replacements, especially as the Benzie and Strain paper seemed to use TPTZ instead of potassium ferricyanide and TCA.
Please let me know if this can be done. Thank you.
You can mail me to rakeshb96@yahoo.com ill send you the reference. And there are more than 10 methods with different chemicals and principles for every assay. You can stick on to any of the protocol according to the availability of chemicals with you
@@Dr.Rakesh.B can I get the reference too?
Pls check the video i made references for all the assays. Details are mentioned
Sir can I can a book's link, so I can download it. Thanks
I want to know the principle of this assay
Thank you really helpful. 😊
Sir I follow this procedure and my absorbance start from 3.52 n continuously INCREASING... Reached upto 4... is it correct?
Sir mirror ke front me baithe hn
sir share the buffer page of book please
how do you interpret the results?
Could you please provide with the reference for citing purpose.
Please watch a separate video I made in my channel for references. You will find my paper you can download and cite the same. If you did not find mail me I'll send my paper rakeshb96@yahoo.com
Can we do the same for cyanobacterial extracts,sir?
Yes, any extracts you can do
@@Dr.Rakesh.B Okay sir, thank you.
What is the formula for calculating FRAP?
Watch the video everything is mentioned in it
I need a method to measure the antioxidant activity in vivo
And if i use phosphate buffer ph: 7, is it normal or not?
You can use
@@Dr.Rakesh.B thank you so much for the information ^^
@@Dr.Rakesh.B sir, i have one doubt . please clarify that. I have chosen my drug with conc.5,50,100,250,500 ug /ml for FRAP in that 5 ug conc. Itself has more colour which is not readble by UV spec.
So that i have planned to reduced my drug conc below 5ug /ml .if it so am i reduce the quantity of reagents like instead of 2.5 ml of PBS use 1ml of PBS ...?
Not required. Bcoz only your concentration is decreasing, but your total volume will be same. Because you are going to make it up to same volume. So don't have to reduce it
@@Dr.Rakesh.B thanks sir for your immediate clarification.
can u share the calculation part also
Hello sir sir plz when u see this MSG plz reply sir m searching for prap assay invitro method it's my project but m not getting it anywhere sir can u please help me out sir plz
I havent done Prap assay. Sorry
Frap*
Sir I want frap assay(invitro) method in pharmacology research ppr
Sir plz reply soon
Thanku so much sir
Thank you sir!
Sir please could you the reference of this FRAP assay
Please watch a separate video I have posted in my channel for all the references. You can cite my paper
Hello sir please provide DPPH chemical
I as an individual can't provide like that from my lab. Moreover my lab works are over.im not going to lab
Hello, can you add subtitles please?🥲
is this frap or reducing power assay ? i just got into an argument because i followed this method, and the prof. says we need TPTZ to do frap
We also use TPTZ to prepare Working FRAP Reagent in FRAP assay
thankyou sir.
Thank you!
Sir we have to prepare different concentrations of plant extracts ??
I had taken data for different concentrations (100-1000 ug/ml) for my four samples.... And the absorbance is increasing for every concentrations... How will I decide.... Which concentration give best results?
And how I will conclude all this data on my results.... It's very confusing... There's very less video available in frap assay... Help me to clear my doubts🙏
Thank you so much.
Thanks
hello sir. can you speak little bit slow to understand
Sure Ok. It was edited to cut short time duration. You can reduce the speed while watching in TH-cam settings