Estimation of Total Carbohydrates by Phenol-Sulphuric acid method//Microplate method//Standard graph
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- Опубликовано: 13 окт 2024
- In this video you will learn the complete procedure, principle of estimating total carbohydrates present in given sample and how to analyze the results by plotting standard graph.
This experiment is performed using microplate (96 well) method. It can also be incorporated for normal spectrophotometry method with cuvettes.
❤very good explanation
Understood thoroughly sir... Very helpful video
Thank you
Very clearly explained🎉
Sir... Can you make video on lipase assay by pH meter method
Thank you so much for this video sir...its very helpful for me to do analysis...can you please post videos on estimation of vitamins both water and fat soluble.
And one more doubt ...plz say about preparation of standard solutions used in vitamins estimation.
which method is most suitable for green microalgae?
bro you are rock yara. your videos are very informative for the researchers... love from University of lahore from pakistan. ;)
Thank you very much
Kindly mention the protocol you followed for the estimation of carbohydrates coz we must have to mention the reference of that protocol in our thesis
Thanks again
Love from National Institute of Genomics and biotechnology Islamabad, PAKISTAN ♥️
what should be ideal temperature of water bath ?incubation is done in dark or in presence light ?please describe slowly .
After Adding Sodium Carbonate.. Vaporw are stopped.. what you have added that the solution is cleared like water??
Sir instead of going for water bath can we autoclave the mixture? Pls reply sir 🙏🏻
No
Do i need to hydrolyse the glucose standard also for 3 hours? Please reply
thank you so much sir. can i use same procedure for the estimation of Total carbohydrate of in insect sample
Yes, you can use.
Sir when I was doing going carbohydrate estimation of the plant Leaf sample.I took 2 sample plant dried leaf sample and fresh leaf extract sample,when I was doing that it showed alage concentration difference .The dried sample had more carbohydrate concentration but in fresh plant it was less.can you tell me a few reason why it was.so that it will so use full for my final year project.waiting for your reply sir😊
The fresh sample has lot of water content its majorly made up of water (75 to 90%) so u got less value (diluted)
While dry sample you have evaporated all moisture and therefore when you take 1.0g of fresh sample its less and 1.0g of dry sample more cells are there u get concentrated value
I also want to determine carbohydrate content in dried leaf sample please can you tell how to treat that solid substrate for further analysis.
Sir what is the amount of stock that you have prepared in this experiment
Can we use Sodium hydroxide instead of Sodium carbonate??? 🤔
Your video are very helpful sir👍
Thank you for such a nice demonstration !!
Can you pls make video on estimation of riboflavin (Vit B2) UV vis spectrophotometeric method
Thank you. Sure :)
@@Dr.Rakesh.B I need your advice on a matter related to this method. Can I contact you?
Sir please show us how to use ELISA plate and how to load in instrument. And if our college does not have this facility how we should take. Absorbance. Please guide.
Will adding random (until effervescence disappears ) Sodium carbonate won't effect the reading ? Thank you
No
@@Dr.Rakesh.B Thank you so much, Sir
some of the samples have initial colour. Is it not create a matrix effect when taking the uv ?
How much sample I have to put in the micro plate reader
Sir please make an video on total reducing sugars and free amino acids...and you are doing great job
can i use this method for analysis of carb in fish muscles. and if water bath is not available any other option? please suggest
Yes you can use. Apart from this you have anthrone method which is more common for animals. But no issues you can use any of the two.
If there's no water bath you should heat it using bunsen flame or heating mantle something like that. Because heating is mandatory step so that the cell disrupts and all carbohydrates are released.
Sir, how to prepare the standard curve... Sir in this if v prepared working standard solution from stock solution, then how to calculate the concentration of glucose..
I am not asking in unknown sample... My question in standard curve table how to calculate concentration of glucose
sir mujhe seed ke water extract me se carbohydrate estimate krna h to kya hm2.5 N HCl ki jgh water le skte h
You said not to put dried sample in high temperature because proteins are heat sensitive , but here you are doing carbohydrates ?
Yes, you are exactly right. Principle for estimation of Carbohydrates is because of high temp all Carbohydrates gets hydrolyzed and that turns into furfural complex in presence of acid. So definitely all Carbohydrates gets broken.
I want to determine total carbohydrate from groundnut seeds
But have defatted sample(remain after oil extraction) can i use that sample for carbohydratr estimations
Yes, if your sample was not treated with any acids or high temp above 100 degrees then definitely you can. If you had treated with acids or high temp there's a possibility that carbohydrates might have got denatured
@@Dr.Rakesh.B yes i extracted oil by soxlhet method (hexane)
Thanks sir
Plz koi bta do caratinoied ke hexane vale test me absorbance kitne wavelength pr lete h?
Please make the video on calculations
Sir, my sample is a brown coloured beverage.. after adding phenol nd sulfuric acid, its colour becomes very dark whose OD goes beyond 3.. Even after diluting 100 fold resulting OD is high... what should j do?
There's no harm in diluting it even 1000 or several times. Because at the end you are gonna cross multiply. So dilute even more. Take very less and make.it.up to huge volume say for eg. Take 10ul make up to 1 ml
@@Dr.Rakesh.B sir after finding concentration of sugar using equuation in ms excel.. the concntration coming is 90 microgram per ml but if i multiply it with 400 dilution factor it comes 39,000 micrgram per ml. Which is not possible nd too high for my sample.. answer should come in percentage below 20 %. Can you please help me sir
Mera Bhi result yahi aaya phenol aur sulfuric acid add krne par brown color hogya
Can you please make a video on estimation of reducing sugar from Plant extract by dns method??
Sir,
Is total carbohydrate and total sugars the same?
Yes it's same.
Plz make a video on calculations also
Absolutely
Thanks man! You speak so fast...
Its edited to cut short the time. Sorry for the bad editing. Reduce the speed in RUclips settings.
can i get the paper for hournal thi by this week
Please watch my video in my channel where I have mentioned about my papers. If u did not find it mail me to rakeshb96@yahoo.com I'll send you my paper you can cite my paper
Do you have the protocol for this method? Help me, please
Yes. Pls do watch the video i made in my channel for all the references
Hi, do we need to hydrolyse the Glucose standard also for 3hrs? or only sample is hydrolysed?
Did you get the answer
Did you get the answer
Can you please upload a video on lipid extraction method with calculation.
Please say glasswares used in this practical
Test tubes, tips, micropipette, beakers and reagent bottles
Make a vedio on estimation of Total sugar, reducing sugar
can i use sucrose as standard ? glucose is not available
@@Dr.Rakesh.B thanks a lot for reply.. i have mailed some queries.. please find out some time to reply to those.. thanks..
OD ke time me auto zero kisse Krna h
Thank you
Kindly mention the protocol you followed for the estimation of carbohydrates coz we have to give reference of protocol in our thesis
Thanks
Love from National Institute of Genomics and biotechnology Islamabad, PAKISTAN ♥️
Sir i had a powder form of sample... Can I follow same protocol of urs or some other method.
Sir, at wat temper set up v need to keep for 3 hrs
@@Dr.Rakesh.B 250 mg/ ml std stock solution and from this
10 ml/ 100 ml working standards solution
Sir, I planning my experiment tom... Sir, can u teach me... How to calculate concentration of glucose
In the table...
Very one says by c1 v1 formula.. But I tried but I unable
@@Dr.Rakesh.B thank u sir for ur needfull reply....
Sir can u tell how to calculate the things... Only calculate part..
Sir, if I take 0.1 ml glucose solution.. Then the concentration of glucose will be 100 mg ...
Is it right sir
Make the video on amylose and amylopectin
The calculations are difficult , I did it in different way , but my results are not right ,
Bro can you please Give the link of research paper. For PhD purpose 🟥
I have made a separate video how to access my papers for citations. Please watch that video from my channel.
great
You speak too fast Dude. Slowdown to allow us catch up
Please speak slowly
Very fast voice
Apologies. Editing was improper. Reduce the speed in RUclips settings while playing the video
Itna fast Kyu bol rahe🤔🤔