Sample Preparation for Fatty Acid Composition Analysis_Fatty Acid Methy Ester (FAME) Preparation-GC

[@RFofficial6067]

That is an excellent explanation. I don't have a single doubt about FAME preparation. thanks a lot

[@MicroChemsExperiments]

Good to know. Thanks for staying with us.

[@pushtibhai3857]

Hi, Rushikesh Funde, did you find the fatty acid composition after analyzing in GC? TIA.

[@bijaychhetriconceptofzoolo1886]

Which article you are following to explain the GC process?

[@abigailabu9299]

Well done guys, this is very clear. How do I get the path 2 of this analysis and the reference please

[@akankshasharma3755]

Good n successful method. .. I have done this with flaxseed oil...

[@MicroChemsExperiments]

Well done

[@nnakweJanefrances1414]

hi, would please tell me about your result, thank you.

[@nnakweJanefrances1414]

I am working on manual extraction FAMEs with Rapeseed oil and sunflower oil. Would you please share your knowledge with me about what you did the flaxseed oil? thank you.

[@loveforyou201008]

Thanks for excellent work. May I ask some information? what's the column in GC instrument? I'm planing to use CP-TAP-CB for triglycerides (CP7483). Is it posible to use this protocol? what is the set up condition of GC? Thank you so much

[@alperen1062]

Thanks a lot. How long can I store and reuse methanolic koh and methanolic hcl solutions in a cold, dark place?

[@MicroChemsExperiments]

You can store for 1 month

[@MuhammadIbrahim-ty9bd]

Thanks for all lab testing method.your lab testing channel is best.Dear, please perform Euric acid in fat.

[@MicroChemsExperiments]

Noted. Shortly

[@satishagrawal2700]

Nice

[@itizwhatitiz9560]

Experiment seems to have worked with Flaxseed Oil sample. Good video!

[@jualitafaradhaachnasya9463]

what if the sample is extracted culture media, is it still necessary to do saponification? if it is, what is the max temp? thanks for this amazing vid!

[@chamarakumara9222]

Please can you give 2nd part of this video.. thank you

[@MicroChemsExperiments]

Sure. Soon

[@HishaniPrashar]

it's 2 nd part is uploaded

[@sushmithaj4099]

Thanks for the wonderful explanation. Sir how to calculate the percentage of the components if the calibration curve is not provided. Is there a way out for that as I need to calculate the percentages of fatty acids in rice bran oil. I do not have a calibration curve so can u help me out in this issue. Thank you

[@MicroChemsExperiments]

We will upload video soon about the calculation. Stay with us

[@sushmithaj4099]

@@MicroChemsExperiments It will be really helpful please upload soon possible

[@MicroChemsExperiments]

@@sushmithaj4099 Okay

[@crazyash1000]

Very well explained! Thanks! But i have a doubt regarding aoac method using methanol and h2so4. Here methanol and h2so4 are added with oil and heated at 80 *C for 2hrs. Then hexane is added and it is seperated. Kindly let me know if this is also acceptable method? Thanks

[@MicroChemsExperiments]

We added HCl as a catalyst, not H2SO4. This is laboratory validated method with slight change of original AOAC method. You can use this method for fatty acid analysis.

[@01abhishekanand58]

Can u share any journal article, which refer for this procedure of fame

[@MrLongKR]

Hi. Which method is used for this experiment?

[@MrLongKR]

hi. is the method only designed for oils and fats or its also possible to use this with different samples eg. sausages, meat?

[@MicroChemsExperiments]

This method is only applicable for oils and fat. For other sample, extract oil first and do the same as done in this video using the extracted oil

[@SarkarMuheedinHama]

Thanks for such a clear explanation, But I have a question Can we use polyethylene glycol 400 instead ?? Appreciate your efforts

[@MicroChemsExperiments]

No, you can't

[@md.riadulhoqehimal616]

Great and thanks a lot.. Can you plz share the second part of the video? Eagerly waiting for it...❤️❤️

[@MicroChemsExperiments]

Will upload soon

[@md.riadulhoqehimal616]

@@MicroChemsExperiments Hey, I would be really greatfull if i have linkined/Facebook/what's app profile of you..

[@che5764]

for pure biodiesel, do we need to do these steps since biodiesel is already a methy ester?

[@firdauskhan6154]

Once we are done preparing it...for how long it can be stored for the further analysis on GC??? PLEASE REPLY

[@MicroChemsExperiments]

Analyse as fast as possible (within 24h)

[@rohitrajput8241]

Please learn the further stem Sample inducting in GLC Instrument

[@MicroChemsExperiments]

Ok

[@MrLongKR]

Hi. which reference did you use for this procedure? Thank you~

[@MicroChemsExperiments]

This method is taken from different journals and modified and then internally validated in our laboratory

[@shafiqahnabila5769]

Can you tell which reference journal that you reffered? Cause i need to write on my research

[@01abhishekanand58]

Yes please share the journal article

[@kembarasembang497]

Sir, how to calculate oil yield. I will do the extraction, acid catalyzed and base catalyzed derivatization

[@MicroChemsExperiments]

After derivatization, GC analysis is needed. We will make GC analysis video later. Stay with us

[@kembarasembang497]

@@MicroChemsExperiments thank you so much sir. 😍

[@nivedhitharamesh5936]

Sir ... how to take micron amount of Micro FAME mix (supelco) out of vials?

[@MicroChemsExperiments]

Use micropipette or auto-sampler

[@zeinasul]

Thank you for your video, please how can I get fatty acid composition in milk by hplc method.

[@MicroChemsExperiments]

Not possible. You need to use GC

[@pushtibhai3857]

Hi, I faced a liitle problem here. When I heat the oil and methanolic KOH mixture at 100°C in water bath, it dried out. No clear solution obtained. How can I fix this problem? Thanks in advance!

[@MicroChemsExperiments]

Set condenser properly with cold water

[@mariamshakeel468]

can you tell about the article from where you are following this protocol

[@MicroChemsExperiments]

Try to find from journals

[@epalanisamiepalanisami6863]

Ok .In Boron tri fluoride method which is better methonolic borontrifluoride or ethyl ether complex of boron tri fluoride ?

[@MicroChemsExperiments]

Methanolic BF3

[@epalanisamiepalanisami6863]

Thank u but Iam use ethyl ether complex of boron trifluoride any problem it?

[@epalanisamiepalanisami6863]

Thank you .In boron trifluoride method l use ehtyl ether complex of boron tri fluoride becase low cost .Any difference between methanlic complex of boron tri fluoride and ethyl ethet complex of boron tri fluoride?

[@MicroChemsExperiments]

Using methanolic boro trifluoride is mandatory. Because methyl group from methanol will facilitate the methylation to form fatty acid methyl esters

[@epalanisamiepalanisami6863]

Thank you very much

[@MicroChemsExperiments]

@@epalanisamiepalanisami6863 You are welcome

[@akankshasharma3755]

Sir now can u plz tell me ... how can I done system sustibility for this ... ?? And also about calibration mixture from above method..?? Reference of this particular method

[@MicroChemsExperiments]

We will upload video for your concern topic later. Stay with us

[@deepasamnani7465]

I want a validation procedure for this method.

[@ak-yh1jx]

Do you have a link for sodium sulfate anhydrous? What will happen if we do not use it ?

[@MicroChemsExperiments]

Sodium sulfate is needed to remove trace water from the extract but you can do the test without using it too

[@julietorsinime]

thank you good video, injection in GC system analysis

[@MicroChemsExperiments]

Will upload soon

[@rajanpatel3228]

hi, what if I heat the flask using heating mantle instead of water bath. Would it be any problem?

[@MicroChemsExperiments]

No major problem in result. You can also use heating mantle

[@godagamagds3820]

instead of n-hexane, can we use methanol? because the column which we use is DB-Wax @MicroChem's Experiments

[@MicroChemsExperiments]

No. Methanol will not be appropriate.

[@godagamagds3820]

@@MicroChemsExperiments i am planning to use DB-FFA column and it is high polar column and hexane is non polar so is it gonna be a problem for my FAME anaylse ?

[@grebulocities8225]

​@@godagamagds3820 Not at all - polar columns are not damaged by nonpolar solvents. The hexane solvent peak will appear very early and will not interfere with your FAME analytes in any way. The reason you need a nonpolar solvent is so that you can extract the (fairly nonpolar) FAMEs from the (very polar) glycerol, unreacted methanol, salts, soaps, and water that get produced when you perform the transesterification reaction in the first part of the video.

[@Eazycharts]

Why we add methanolic HCl?

[@23darshana]

Hi. Can you pls let me know why both base catalyst KOH and acid catalyst HCl are used in this method?

[@MicroChemsExperiments]

KOH is used for the saponification of oil. HCl is used to facilitate the methylation to make Fatty Acid Methyl Ester

[@23darshana]

@@MicroChemsExperiments ok. Thanks.

[@23darshana]

can i get any reference to this method if possible?

[@MicroChemsExperiments]

@@23darshana no. Its a lab validated method

[@shahriarislam1733]

which method is this?

[@mirfazlerabbi9011]

In this experiment, there is no need any type of standard?

[@MicroChemsExperiments]

Yes. Standard is needed during the analysis of sample in Gas Chromatograph

[@mirfazlerabbi9011]

@@MicroChemsExperiments which standard is needed? What is the name? And how the preparation procedure please?

[@pushtibhai3857]

Can you please tell me the final concentration of FAME in this method? Thanks in advance.

[@MicroChemsExperiments]

Concentration can be known by analyzing in GC

[@izzatishrrrr]

what is the use of anhydrous sodium sulphate?

[@MicroChemsExperiments]

It is used to absorb and remove trace water molecules from the extract

[@ovaneschakoyan6584]

So that way we methylated all fames c4-c24, that is the best option? Thank you in advance.

[@MicroChemsExperiments]

The best method of methylation is the one which is catalyzed by Boron Trifluoride. But in this video we use acid catalysis technique which is cheaper and simple.

[@dipalimalvekar2472]

How one can prepare animal tissue sample for FA analysis by GC

[@MicroChemsExperiments]

Same procedure

[@md.riadulhoqehimal616]

Could you plz share the chromatographic condition of this method?..

[@MicroChemsExperiments]

We will make video later

[@vikashganvit3548]

Hello ma'am Which colume your use for analysis ?

[@MicroChemsExperiments]

TG Polar

[@vikashganvit3548]

@@MicroChemsExperiments thank u ma'am

[@MicroChemsExperiments]

@@vikashganvit3548 you are welcome

[@vikashganvit3548]

@@MicroChemsExperiments the process will be same for different vegetable oil?

[@MicroChemsExperiments]

@@vikashganvit3548 Yes. Same for all kinds of fats and oils.

[@epalanisamiepalanisami6863]

Can we used sulphuric or nitric acid otherwise hcl

[@MicroChemsExperiments]

No. you must use HCl

[@epalanisamiepalanisami6863]

Thankyou very much . your experiments more useful

[@yahitajam7944]

How much gram of palm raw oil or litre of palm raw oil need to be injected in GC-MS machine?

[@MicroChemsExperiments]

Why would you inject raw oil?

[@yahitajam7944]

@@MicroChemsExperiments i want to analyze Fatty acid profile of palm raw oil. How do we prepare a sample for GC-MS analysis ? Can we directly inject raw oil in a GC-MS or we have to prepare ester before injecting into the GC-MS. *Sorry for my bad English*

[@anfelbellebcir9002]

@@yahitajam7944 i think you have to prepare before the injection (like the video)

[@dhruvamulani3084]

Can you share sample prepration reference name?

[@MicroChemsExperiments]

It is a Laboratory developed in-house method

[@DurayStuffCrafts]

Sodium sulphate anhydrous why we put this in syringe ???

[@MicroChemsExperiments]

To absorb water/moisture from the extract

[@ovaneschakoyan7861]

extraction time is it not too much?

[@MicroChemsExperiments]

Extraction should be faster

[@aswinkumar8016]

Part 2 ????

[@MicroChemsExperiments]

Please wait for the part-2

[@aswinkumar8016]

@@MicroChemsExperiments okay...

[@MuhammadIbrahim-ty9bd]

Part.2

[@MicroChemsExperiments]

Will upload soon

[@aswinkumar8016]

Derivatization

[@MicroChemsExperiments]

Yes. You are right

[@aswinkumar8016]

Pls show us the derivatization of Fatty acid methyl esters using BF3 or BCl3

[@MicroChemsExperiments]

@@aswinkumar8016 Ok. We will make video later.

[@aswinkumar8016]

Ok.. thank u... So much... For ur reply

[@MicroChemsExperiments]

@@aswinkumar8016You are most welcome

[@tallapellyvikas6097]

Can you cite the reference of this derivatization procedure

[@MicroChemsExperiments]

It is a inhouse laboratory developed method

[@tallapellyvikas6097]

Thank you so much it is very effective and easy..👍

[@TehseenKhalid-c2s]

Can we use this method for fatty acid analysis in semen?

[@MicroChemsExperiments]

Yes. You can

[@afeeqafnani9690]

it's already one year now, no part 2 yet

[@MicroChemsExperiments]

Soon