thank you so much sir....your videos are very useful for research work......if possible please make a video on malondialdehyde and lipase of flour samples
Sir.. total volume from: stock+D/w = 1000 micro litres (or 1ml) Or stock+D/w = 500 micro litres (or 0.5 ml) ... You told to make , total volume should be 500 micro litres (or 0.5 ml). Is should be 1000 micro litres (or 1ml)?? Please reply
Sir, I'm doing research on pigment extraction using different organic solvents from flowers . Can I use the pigment extracted directly for phenol estimation without drying to powder form?
Sir will you plz tell me that if i am preparing Gallic acid reference standard solution for standard curve preparation for phenolic content determination then if i have taken 10mg Gallic acid in 10 ml distilled water and then taken 6 concentrations from that 10mg dissolved in 10 ml distilled water solution for taking absorbance for making standard curve then is it correct way of making standard curve . And sir there is no need of adding working solution in standard solution for making standard curve?? Pls help this is for my research work
There are 2 methods, you can do the method i said its easy. The other one with working solutions and all is bit complicated. Yes 5 concentrations are enough. You r doing it correct. All the best 👍
When we are taking the absorbance in Spectrophotometer then if we are taking samples and all the chemicals by multiplying 10 to the volume you suggested in this video then is it correct or not?
I am using choline chloride:urea deep eutectic solvent for extracting phenolic compounds. The problem is that when I add folin reagent to (gallic acid+deep eutectic solvent) standards prepared, it form precipitate or colloids which settle down at bottom after some period of time. It doesn't happen when I use water as solvent. Do You know why it i happening?... Also I use 7.5% NA2CO3 for estimation.
Stock soln of Gallic Acid and water is prepared, now I hv to prepare standrd solution for stndrd curve, so may i know wht compunds we use to make standard curve other than stock solituon?? Please reply...means alot
Sir, how to make dilution of folin ciocalteu reagent (from sigma Aldrich) , it's written 2N on the bottle ... What is the desirable conc. For standard preparation for phenolic estimation.?? Thank you
1:1 or 1:2 both are ok. I preference 1:1. For standard you need to do trial and error. We can't follow strictly or I can't just say some concentrations.
@@Dr.Rakesh.B sir, I have tried making standard for phenolic estimation, i used all the reagents and their conc. in the same ratio as you have mentioned here, but every time getting high absorbance value while making standard.... How to resolve this... 🙏
Thanku sir for this video...but I m confused how should we decide the exact concentration of our sample in methanol or ethanol which needs to be estimated. Kindly help
After evaporating say for eg. U get 0.01g of extract that is 10mg. Add 10 ml of solvent. Now ur conc is 1mg/ml. Like this based on what u get u can add to get 1mg/ml, 10mg/ml 5mg/ml. Everything is crct
I have seen your DPPH, phenolic,methanolic extraction of plant sample and FRAP.All are very good.How beautifully you explained everything.It is commendable.I am also doing my work on plant sample. If possible could you make a video on SDS-PAGE. Especially on sample preparation using plant sample.Thank you
@@Dr.Rakesh.B Thank you, sir. Sir if you don't mind can you please share the references for phytochemical analysis (quantitative estimation) & for methanolic extract preparation?
Can you please tell me how to make a plant extract? I mean can we use oven-dried plant material at 60 degrees C to make an extract? If so, what solvent can we use?....any help would be appreciated.
Sir, you added 1 gm of gallic acid to 100 ml of distilled water. Then how can the concentration of 1mg/ml make? please explain this and rectify me if I am wrong here.
Do you have any video in which you explain the calculation for the analyzed compound's concentration based on the sample amount I used? This is because I should consider the amount of plant same I used for analysis as well as dilution factor etc..
1:1 dilution i did. I have read papers which showed 1:10 dilution also but it depends on the concentration of the chemical , purity and brand. I suggest you to do 1:1
Sir when I apply in ethanolic extract there are some turbidity problem in my different samples and spectrophotometer increase the O.D. how to deal with that
You can get my papers in research gate or Google Scholar. I have made a separate video on how to get my papers for references. Please watch thT from my channel.
Sir, I have a question, I have standard graph of pigment prepared in Petroleum ether. But for other assay, I have to solubilize in another solvent like methanol, In that case if I dry Petroleum ether extract of volume 1ml with conc 50ug/mL and solubilize in 1ml methanol will the concentration be same but OD will vary? Please send me some suggestion
Actually you are supposed to use methanol only. Thats logically correct. In some cases we do slight modifications like this if we are having shortage of chemicals
@@Dr.Rakesh.B thanks 4 reply but sir concentration per ml or per 100ml would be changed. during calculation after calibration curve which concentration will be taken into consideration with respect to only gallic acid or total solution concentration
It's mentioned in video if I'm not wrong. Kindly watch without skipping. Or watch videos of other biochemical assays to understand blank and control concepts
Sir, As I did same but I got precipitate in the blank as well as In all samples after adding Sodium carbonate Do u know the reason if so kindly tell how to prevent from this My 2nd question is If we take 1 ml FCR reagent then we have to simply add 1 ml of distilled water or we should dilute it 10 times
I took 4, 6,8,10,12,14,16,18 ul from stock solution (0.5mg/ml ) n draw standard curve 1 got 10.40 of my Sample after applying y= mx+b I took 10 ul from Sample (which is having 10 mg/l) So my question is what will be the unit of my Sample of which I got value 10.40 Will it be 10.40 ug/ml ? Or does it mean that in 10 ul of my Sample there is 10.40 ug ?
Thank You very much Dear Brother! It helped me alot!
Can you please make a video on making standard graph of these values?
Thanks a lot. i was struggling to understand my lab work which requires total phenol estimation of Bacopa. Your video helped me
Your welcome:)
thank you so much sir....your videos are very useful for research work......if possible please make a video on malondialdehyde and lipase of flour samples
Thanks for the video Sir.. can we use plant extract dissolved in DMSO instead of methanol/ethanol for the experiment?
Yes you can 👍
Sir.. total volume from:
stock+D/w = 1000 micro litres (or 1ml)
Or
stock+D/w = 500 micro litres (or 0.5 ml)
...
You told to make , total volume should be
500 micro litres (or 0.5 ml).
Is should be 1000 micro litres (or 1ml)??
Please reply
Thankyou very much sir...
But I have one doubt do we have to add Gallic acid to the plant extract as well ??
No no gallic acid will not be added.
@@Dr.Rakesh.B thankyou sir for your quick response🙏🙏
Sir can we use Chloroform extract.
Sir, I'm doing research on pigment extraction using different organic solvents from flowers . Can I use the pigment extracted directly for phenol estimation without drying to powder form?
Yes, you can if you know the concentration of the pigment
Refrence for the said experiment any paper mg galic acid equivalent per 100 g for total phenolic plzz mention
Download my paper, I have given details in a video in my channel for papers
Sir will you plz tell me that if i am preparing Gallic acid reference standard solution for standard curve preparation for phenolic content determination then if i have taken 10mg Gallic acid in 10 ml distilled water and then taken 6 concentrations from that 10mg dissolved in 10 ml distilled water solution for taking absorbance for making standard curve then is it correct way of making standard curve .
And sir there is no need of adding working solution in standard solution for making standard curve??
Pls help this is for my research work
There are 2 methods, you can do the method i said its easy. The other one with working solutions and all is bit complicated.
Yes 5 concentrations are enough. You r doing it correct. All the best 👍
Plz help how extraction of free and bound phenolic compounds
Can you please tell the calculation to estimate the total phenol with the dilution values
Very nice information sir 👌👌👍
Which instrument is using for spinning
Can I get a graphical explanation and results of what u performed in this video?
When we are taking the absorbance in Spectrophotometer then if we are taking samples and all the chemicals by multiplying 10 to the volume you suggested in this video then is it correct or not?
Please reply because tomorrow I am going to do phenolic estimation
It is correct
You are the first one whose videos need speed less than normal to play😂....But big thank you for imp information 😊
Yeah unknowingly during editing speed was increased to cut short the video duration. Apologies for that.
Thank you 🙏
Sir ready made FC reagent available ho toh Direct use kare ya kaise dilute kare?
Videos are very useful. But if possible try to mention your protocol reference publication in the description
@@Dr.Rakesh.B ok
I am using choline chloride:urea deep eutectic solvent for extracting phenolic compounds. The problem is that when I add folin reagent to (gallic acid+deep eutectic solvent) standards prepared, it form precipitate or colloids which settle down at bottom after some period of time. It doesn't happen when I use water as solvent. Do You know why it i happening?... Also I use 7.5% NA2CO3 for estimation.
thankyou so much for the video, but can I ask for some clarifications, if the absorbance is at 615 or 650nm?
650nm
Stock soln of Gallic Acid and water is prepared, now I hv to prepare standrd solution for stndrd curve, so may i know wht compunds we use to make standard curve other than stock solituon?? Please reply...means alot
It was a great illustration. Thank you sir.
Please make a video on quantification of saponins and lignins of plant extracts
Sir, how to make dilution of folin ciocalteu reagent (from sigma Aldrich) , it's written 2N on the bottle ... What is the desirable conc. For standard preparation for phenolic estimation.??
Thank you
1:1 or 1:2 both are ok. I preference 1:1.
For standard you need to do trial and error. We can't follow strictly or I can't just say some concentrations.
@@Dr.Rakesh.B sir, I have tried making standard for phenolic estimation, i used all the reagents and their conc. in the same ratio as you have mentioned here, but every time getting high absorbance value while making standard....
How to resolve this...
🙏
Reduce the std concentrations or increase their volumes
Thanku sir for this video...but I m confused how should we decide the exact concentration of our sample in methanol or ethanol which needs to be estimated. Kindly help
Sir kindly reply
After evaporating say for eg. U get 0.01g of extract that is 10mg. Add 10 ml of solvent. Now ur conc is 1mg/ml.
Like this based on what u get u can add to get 1mg/ml, 10mg/ml 5mg/ml. Everything is crct
Watch how to prepare methanolic extract video from my channel for more information
@@Dr.Rakesh.B ok sir...I m clear now
Sir please share us the paper u follow.,the way u explain is easy to understand n cleared my doubt...😁
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
I have seen your DPPH, phenolic,methanolic extraction of plant sample and FRAP.All are very good.How beautifully you explained everything.It is commendable.I am also doing my work on plant sample.
If possible could you make a video on SDS-PAGE. Especially on sample preparation using plant sample.Thank you
Thank you. Will try that I'm not sure if I can because my lab works are over. May be in future I'll try it.
Sir maine aapke protocol se phenol test Kiya tha aur aapke paper ko site kiya tha ....mera paper published ho gya
Congratulations 🎊 👏
How to determine polyphenols and oxidation in curcumin
can you please make a video on the extraction of plants(halophytes) biomass composition (cellulose, hemicellulose and lignin
Thoroughly understood sir... It's very helpful for understanding my research work
Hi sir thank you for the video a doubt can I increase the incubation time ?
Yes
@@Dr.Rakesh.B Thank you, sir. Sir if you don't mind can you please share the references for phytochemical analysis (quantitative estimation) & for methanolic extract preparation?
Mail me rakeshb96@yahoo.com I'll send my paper u can cite the same
@@Dr.Rakesh.B okay sir i will thank you so much.
Sodium carbonate will be added in blank or not? Please reply
It will be not be added in blank. But it will be added in control
@@Dr.Rakesh.B when absorbance of sample is taken then what will be used as reference in spectrophotometer? Please reply 🙏
Can you please tell me how to make a plant extract? I mean can we use oven-dried plant material at 60 degrees C to make an extract? If so, what solvent can we use?....any help would be appreciated.
Blank for gallic acid is Distilled water and FC reagent and for sample is same
Sir, you added 1 gm of gallic acid to 100 ml of distilled water. Then how can the concentration of 1mg/ml make? please explain this and rectify me if I am wrong here.
Take 100 mg in 100 ml to make 1mg/ml concentration
Sir, if we are making 1mg/ml stock of gallic acid and taking 400ul from it then how to know the conc. of gallic acid in this 400 ul .?
Thank you
Hello sir. If I want to amke 1mg/ml conc Gallic acid, is it right to dissolve 0.5g of Gallic acid in 50ml of water
sir can you explain how to obtain answer in terms of mg GAE/g sample. GAE= Gallic Acid Equivalent
Sir can we use Gallic acid for tannic acid estimation also ?
Yes you can
Thank you sir
Is Folin ciocalteu and folin denis reagent both are same?
Well. Explained.
Sir can u make a video on cellulose estimation ..
Sir.. Can you suggest to me any standard book for quantitative estimation of phytochemical
Thank you so much, you are my oasis...
Do you have any video in which you explain the calculation for the analyzed compound's concentration based on the sample amount I used? This is because I should consider the amount of plant same I used for analysis as well as dilution factor etc..
Thank you :)
Nope. I just briefly explained them in standard graph video. Will do back calculations in separate video soon
Thank you very much sir. It helped me a lot. You are the best
Sir if we use hexane plant extract of 1mg/ml conc then what would be blank either hexane or methanol?
It will be hexane. Remember always the solvent which you have used for preparing plant extracts will be taken.
thanks alot sir how many times FC reagent is diluted is it ok to use 1:10
1:1 dilution i did.
I have read papers which showed 1:10 dilution also but it depends on the concentration of the chemical , purity and brand.
I suggest you to do 1:1
@@Dr.Rakesh.B thanks alot
Can we use the same amount of catechol instead of Gallic acid in same concentration..?🙏
Yes u can
Sir... Any equation for total phenolic content estimation??..how we will calculate the content?
Watch the standard graph and calculation video from my channel. That will help you calculate
Ok
Can you please upload video on antioxidant activity by ABTS?
Ok
Sir can I replace the Gallic acid with the extract of the plant?
If I add 5ml of 1mg/ml of stock solution and add 95ml of dw, does It make 50microgram/ml of the gallic acid solution ?
Hello sir, basically the absorbance should remain below 1, mine is going above one.. please tell me what could be the reason for this?
Can i know reference for this procedure
Mail me rakeshb96@yahoo.com I'll send paper
Thanks alot sir, your video really give me some ideas for doing TPC for my plant extract! it is really informative video.
Sir when I apply in ethanolic extract there are some turbidity problem in my different samples and spectrophotometer increase the O.D. how to deal with that
I also experienced the same problem
7.5% sod. carbonate k liye 7.5mg dissolved karna padega 100ml main
How much sodium carbonate do you add in the tube ?
Sir, plz tell me how to extract phenol from food before going for fc method
I haven't done phenol. Only phenolics i did
@@Dr.Rakesh.B yes sir i meant the same, it is a cookie sample.how should i take extract
Powder it and make methanolic extract like i explained in one of my video
Sir May i have your paper please ? For citation? Or what should i write for this in my paper as citation?
You can get my papers in research gate or Google Scholar. I have made a separate video on how to get my papers for references. Please watch thT from my channel.
@@Dr.Rakesh.B please give me the link of your paper or that video showing how to get your paper
Please Can you make video on assay for tannin content
Sir, I have a question, I have standard graph of pigment prepared in Petroleum ether. But for other assay, I have to solubilize in another solvent like methanol, In that case if I dry Petroleum ether extract of volume 1ml with conc 50ug/mL and solubilize in 1ml methanol will the concentration be same but OD will vary? Please send me some suggestion
@@Dr.Rakesh.B thank you so much sir
could you please give me written document of it?
Video is nice, but the delivery speed is supersonic, could be little slow, for easy catch ups
Sir what will be estimated cost of finding out total phenolic content in a drink ?
Do drinks have phenolic compounds?
You can only analyze its not costly all chemicals are cheap. U just need spectrophotometer
@@Dr.Rakesh.B kombucha has phenolic content .. okk thank u so much . I learnt a lot ☺️
why you dilute the methanolic extract with water but not methanol? im confused
Actually you are supposed to use methanol only. Thats logically correct.
In some cases we do slight modifications like this if we are having shortage of chemicals
Can you please make a video on Reducing power effect assay
Sir plz show us how to do anti-inflammatory test of extracts
Thankyou so much for this video... plz make a video on total flavonoid content (TFC) estimation as well
Raki's Just Biology Thankyou so much 😊
Very informative video. Thank you👍
What is the conc of plant extract?
Very nice explanation
hai sir thank you so much for ur video. can you please provide a valuable reference paper for this method? which helps me alot.
Sure. I'll share the link of my published paper you can cite it. It will be published within 10 days then ill make a video and let you all know.
sir, i want to ask that when we added FC, h2o, and sodium carbonate. concentration of galic acid why not changed
@@Dr.Rakesh.B thanks 4 reply
but sir concentration per ml or per 100ml would be changed.
during calculation after calibration curve
which concentration will be taken into consideration with respect to only gallic acid or total solution concentration
Sir what about blank And control
It's mentioned in video if I'm not wrong. Kindly watch without skipping. Or watch videos of other biochemical assays to understand blank and control concepts
@ 8:27 how much sodium carbonate should be added after keeping for 5 to 10 minutes.
Until the effervescence I means the bubbles or foam ceases more than 2-3 g I added
Sir please do videos on estimation of antioxidant enzymes like SOD, peroxidase, catalase and non enzymatic antioxidants estimation
Sir,
As I did same but I got precipitate in the blank as well as In all samples after adding Sodium carbonate
Do u know the reason if so kindly tell how to prevent from this
My 2nd question is
If we take 1 ml FCR reagent then we have to simply add 1 ml of distilled water or we should dilute it 10 times
@@Dr.Rakesh.B ok sir
I took 4, 6,8,10,12,14,16,18 ul from stock solution (0.5mg/ml ) n draw standard curve
1 got 10.40 of my Sample after applying y= mx+b
I took 10 ul from Sample (which is having 10 mg/l)
So my question is what will be the unit of my Sample of which I got value 10.40
Will it be 10.40 ug/ml ?
Or does it mean that in 10 ul of my Sample there is 10.40 ug ?
@@Dr.Rakesh.B thanks
So TPC of my Sample will be 0.1 mgGAE/g extract
By applying TPC = C×V/M
Is it so?
@@Dr.Rakesh.B thnks
For what procedure for bacteria and fungi filteration
I have made a separate video for that in my channel pls watch
OD kitne nm pr lena h sir?
650 nm
Sir make video on plant enzymes sod, pod, cat, apx
If we are getting absorbance more than 3 or 4 what we should do?? Abs should be less than 1 always?
@@Dr.Rakesh.B thank u sir
Sir plx reduce the speed of your talk at the time of introduction
Im sorry for the speed it's edited to make the video short. Try to reduce the speed of the play in TH-cam settings and watch.
If R^2=0.7811 is ok?!! Plz answer me
You need to ask your research guide if he/she is ok then u can.proceed
@@Dr.Rakesh.B thank you sir, plz can you do estimation of tannins by vanilline méthode, 🌸🌸🤍
500microliters in mili liters please mention it!!!
0.5 mL
Thank you so much
Sir can you please mention your gmail
Gallic acid 1gm /100ml ho toh 1mg/ml kaise ?
Absorbance at 650 nm or 765 nm
Sir i totally got this but sir can please slow it down little. Your talking to much faster :)
It has been edited, sorry for that bad edit. Slow down in youtube settings while watching
Sample preparation for bacteria and fungi of phenols
Thank u
Sir thoda slow bola kro
PLEASE TOTAL TANNIN CONTENT PER VIDEO PREPARE KIJI AE NA
Sir i have sent mail requesting this paper for reference.. It would be so great ful if u could send same.
Thankyou.
Suji
Pls make this vedio in hindi
Go slow bro that we can understand well.
Sure. Thank you
Slowly u do sir
why you speak soo fast. Speak slowly bro
Plz speak slow your speed is not normal
@@Dr.Rakesh.B thanku so much for your kind response you are doing so well