Thank you very much for your help. I have a question about 1/ the sodium phosphate product; should I use the monobasic or the dibasic ? 2/ what does the blank contain?
Hi! I was going through the literature on Phospho-Molybdate Assay (Total Anti-oxidant Capacity) and I was puzzled by the formula for the percentage anti-oxidant capacity which is given below %Antioxidant effect =(control absorbance - sample absorbance/ control absorbance)*100 To the best of my understanding, in this assay the anti-oxidant capacity increases as the absorbance increases. This should mean the control sample should have the lowest absorbance and hence the lowest anti-oxidant capacity. It would also mean that if I were to use the formula given above, the numerator would come out as negative. Can we use negative values? Is the formula correct? Or is my understanding of the basic principle of the assay flawed? Kindly help!
Thank you so much, sir, your videos are very helpful and your explanation is very good. Please explain the Hydrogen peroxide and hydroxy radical assay for measuring antioxidant activity.
Sir, is the concentration of the sample (10 mg/ml) for a total volume of 100 microliters or 1100 microliters? I mean, was 100 microliters of 10 mg/ml concentrated sample used or the concentration of the sample made 10 mg/ml for a total final volume of 1100 microliters?
Hello sir l-ascorbic acid is not able to dissolve in solvent like methanol, acetone. Should we make Standards with water even though the plant extract made by methanol.
Pls for the negative control if ur solvent of extraction is not water or ethanol but ethyl acetate can u use it as the base for the negative control .thank u need and ASAP
@@Dr.Rakesh.B pls I'm having difficulty carrying out my anti oxidant assay bcos the UV machine in my sch is like I don't know if I should say manual we have to obtain our calibration curve first
Helo sir, I have a doubt, while we conducting anti oxidant assys, we prepare different concentrations of our sample. But in the case of this assay we take only single concentration. Is single concentration is enough to find antioxidant activity of our sample by phosphomolybdate assay?
sir if i want to see the antioxidant by dpph of fixed oil thn i cannot dilute in methanol ...thn if i use hexane do i need to use hexane in standard for ascorbic acid...do ascorbic acid can be dissolved in hexane or petroleum ether
@@Dr.Rakesh.Bsir, i followed the procedure as you taught. Even though, i am getting absorbance reading in higher to lower value for high to low conc. Which ends with the result negative value. I repeated again. But getting same. Please help to solve this. Waiting eargerly.
@@Dr.Rakesh.B sir, i have three doubt. sodium sulphate , ammonium molybdenum dissolved in distilled water or selected solvent ? How to check the sulphuric acid strength for this assay ? Please describe this part. I am using test tube for phosphomolybdenum method and i have chosen the conc. as follows 5ug, 50,100, 250,500,1000ug/ml.In that i have use my solvent to make up to 1 ml as final volume of my sample volume+1 ml PMR reagent and i followed the steps as like procedure. Is this correct??? Please clarify my doubts which is very needful to me.
@@Dr.Rakesh.B sir, i followed the procedure as you taught. Even though, i am getting absorbance reading in higher to lower value for high to low conc. Which ends with the result negative value. I repeated again. But getting same. Please help to solve this. Waiting eargerly.
Sir could u please tell me the pattern of absorbance (from high conc. to low conc.).... As my absorbance is coming high in high conc. And vice versa.... But in finding the %inhibition , the results r coming negative.... I have repeated this experiment several tyms nd the results r same.... Plz help me.
Hello @sunainamohana5141, could you explain what did you for the control, because sir's suggestion is deleted As the concentration increased my od value increased, but my percentage came negative
I did this method .. I got yellow color it means ... Can u explain what the colour change indicates and how absorbance values should come while reading ??
Sir, please tell me how to prepare ammonium molybdate solution. Because I tried to prepare itu, but ammonium molybdate didn't dissolve properly in water.
@@Dr.Rakesh.B Thank you Sir. The heating process was found to increase the solubillity. I prepared the reagent and run the assay successfully. Since I didn't find the control solution (like DPPH), Is it possible for me to to obtain the IC50 with this assay?
could you please tell us the pattern of absorbance at different concentrations... as m getting high absorbance at high concentration nd vice versa... as a result of which... percentage inhibition is coming in negative.... blank value is coming from 0.035 to 0.045..... i have repeated this experiment 7 times.... can u plz help???
thanku for replying sir.... today i tried again wid d concentrations suggested by you.... bt d outcome was same... high absorbance at high concentration.. nd vice versa.... can u please suggest any solution??
Sir yours explianation was awesome and very helpful. Thank you.
Sir can you plz upload HRBC membrane stabilizing assay...
❤
Ur explaination is great sir. I want to ask , if we compare all the methods of antioxidant activity. will i get same answer.
Thank you very much for your help. I have a question about
1/ the sodium phosphate product; should I use the monobasic or the dibasic ?
2/ what does the blank contain?
Good video, have you worked with the ORAC Assay?
Sir we have made aqueous plant extracts...so how we prepare standard ascorbic acid....in ethenol or in aqueous
Hi! I was going through the literature on Phospho-Molybdate Assay (Total Anti-oxidant Capacity) and I was puzzled by the formula for the percentage anti-oxidant capacity which is given below
%Antioxidant effect =(control absorbance - sample absorbance/ control absorbance)*100
To the best of my understanding, in this assay the anti-oxidant capacity increases as the absorbance increases. This should mean the control sample should have the lowest absorbance and hence the lowest anti-oxidant capacity. It would also mean that if I were to use the formula given above, the numerator would come out as negative. Can we use negative values? Is the formula correct? Or is my understanding of the basic principle of the assay flawed?
Kindly help!
Thank you so much, sir, your videos are very helpful and your explanation is very good. Please explain the Hydrogen peroxide and hydroxy radical assay for measuring antioxidant activity.
Thank you
Sir which method can be used to check the antioxidant activity if water is used as a solvent
Whether colorimeter can be used instead of spectrophotometer?
Yes
tanks.
please can you assist me on how to calculate the TAC of plant extracts in this assay
We, our pharmacy students, are doing project work, so will you do the activities in your lab?
I'm not working in lab anymore
Sir, is the concentration of the sample (10 mg/ml) for a total volume of 100 microliters or 1100 microliters? I mean, was 100 microliters of 10 mg/ml concentrated sample used or the concentration of the sample made 10 mg/ml for a total final volume of 1100 microliters?
Hello sir l-ascorbic acid is not able to dissolve in solvent like methanol, acetone. Should we make Standards with water even though the plant extract made by methanol.
Sir we don't hv water bath.. can we keep in oven at this temperature
Dear sir , can we use di-sodium hydrogen phosphate as an alternative to sodium phosphate?
Yes
Sir what is the Blank we should use for uv microscopy??
The solvent which was used for extract preparation
Thank you sir
Pls for the negative control if ur solvent of extraction is not water or ethanol but ethyl acetate can u use it as the base for the negative control .thank u need and ASAP
Yes you should use the same solvent as your negative control
@@Dr.Rakesh.B pls I'm having difficulty carrying out my anti oxidant assay bcos the UV machine in my sch is like I don't know if I should say manual we have to obtain our calibration curve first
@@Dr.Rakesh.B n I totally have no idea on how to go abt the calibration n what solvent to use to calibrate.thanks alot I hope to get a reply soon
Sir , if we will do this assay in the dilution tubes , then how much volume we should take? Plz guide. Bcz we have colorimeter .
Helo sir,
I have a doubt, while we conducting anti oxidant assys, we prepare different concentrations of our sample. But in the case of this assay we take only single concentration. Is single concentration is enough to find antioxidant activity of our sample by phosphomolybdate assay?
Which one of the following salts is used in this method?
Both of them can be used ? (NH4)2MoO4 and (NH4)6Mo7O24.4H2O salts
I have used 2nd one. Yeah anyone can be used
@@Dr.Rakesh.B Today I did the experiment and got the good results. Thank you very much.
Sir please upload how to do antioxidant enzymes activity by microplate method
sir if i want to see the antioxidant by dpph of fixed oil thn i cannot dilute in methanol ...thn if i use hexane do i need to use hexane in standard for ascorbic acid...do ascorbic acid can be dissolved in hexane or petroleum ether
Seriously I want u as my professor at my college😉
@@Dr.Rakesh.B sir please give your email id for some quieres discussion sir
@@Dr.Rakesh.B I have mailed my doubts sir. Thank you for the quick reply sir.
Superoxide free radical scavenging activity ki please vedio share share kijiye
Sir please tell us how to prepare 28mM sodium phosphate
4mM ammonium molybdate
Watch my video on calculations in my channel
@@Dr.Rakesh.Bsir, i followed the procedure as you taught. Even though, i am getting absorbance reading in higher to lower value for high to low conc. Which ends with the result negative value. I repeated again. But getting same. Please help to solve this. Waiting eargerly.
for how long can we store the phosphomolybdate reagent
Use fresh
Sir, I just want to know wht reference you are using to do this test, please give me the reference link
Pls watch my video on references in my channel. You can download my paper and cite the same
@@Dr.Rakesh.B sir, i have three doubt.
sodium sulphate , ammonium molybdenum dissolved in distilled water or selected solvent ?
How to check the sulphuric acid strength for this assay ? Please describe this part.
I am using test tube for phosphomolybdenum method and i have chosen the conc. as follows 5ug, 50,100, 250,500,1000ug/ml.In that i have use my solvent to make up to 1 ml as final volume of my sample volume+1 ml PMR reagent and i followed the steps as like procedure. Is this correct???
Please clarify my doubts which is very needful to me.
Dissolve is distilled water.
Watch my video on calculations to calculate acid strength.
Yes you are correct
@@Dr.Rakesh.B thank you very much sir for your immediate response.
@@Dr.Rakesh.B sir, i followed the procedure as you taught. Even though, i am getting absorbance reading in higher to lower value for high to low conc. Which ends with the result negative value. I repeated again. But getting same. Please help to solve this. Waiting eargerly.
How much time to incubate at 90 degree..sound is not clear?
90 minutes.
Sir could u please tell me the pattern of absorbance (from high conc. to low conc.).... As my absorbance is coming high in high conc. And vice versa.... But in finding the %inhibition , the results r coming negative.... I have repeated this experiment several tyms nd the results r same.... Plz help me.
@@Dr.Rakesh.B thank u so much sir... Will try again nd focus on minute details
I also have same issue and also want to know about the absorbance of blank. it should be higher than sample or not?? sir kindly anwser it
@@sunainamohana5141 I also facing the same issue. Can you say me the mistake.
Hello @sunainamohana5141, could you explain what did you for the control, because sir's suggestion is deleted
As the concentration increased my od value increased, but my percentage came negative
how to take mmol
sir from which stream you are?
Are you also from Biochemistry?
I did this method .. I got yellow color it means ... Can u explain what the colour change indicates and how absorbance values should come while reading ??
@@Dr.Rakesh.B thanks sir... I will try again..
Sir, please tell me how to prepare ammonium molybdate solution. Because I tried to prepare itu, but ammonium molybdate didn't dissolve properly in water.
@@Dr.Rakesh.B Thank you Sir. The heating process was found to increase the solubillity. I prepared the reagent and run the assay successfully. Since I didn't find the control solution (like DPPH), Is it possible for me to to obtain the IC50 with this assay?
@@Dr.Rakesh.B so if I want to calculate the IC50, I have to plot the concentration vs OD?
Could you please modify Google Translate so that it can be translated into Spanish because I am bad at English.
could you please tell us the pattern of absorbance at different concentrations... as m getting high absorbance at high concentration nd vice versa... as a result of which... percentage inhibition is coming in negative.... blank value is coming from 0.035 to 0.045..... i have repeated this experiment 7 times.... can u plz help???
thanku for replying sir.... today i tried again wid d concentrations suggested by you.... bt d outcome was same... high absorbance at high concentration.. nd vice versa.... can u please suggest any solution??
@@avneetmakkar417 I also facing the same issue. Can you say me the mistake.
souiduim phsphate mono or di phoshate
@@Dr.Rakesh.B شكراا جزيلا thanks
Sir , can u mention calculation part?
@@Dr.Rakesh.B may I have the link of the same