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

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Күн бұрын

Fatty Acid Composition (Fatty Acid Profile) Analysis is a commonly used technique in lipid research and oil or fat test which is very important to check the quality of an oil or fat sample. Analysis of fatty acid composition is usually carried out by gas chromatography (GC). Sample preparation is very crucial for the Fatty Acid Composition analysis. Free fatty acids, present in oils or fats, cannot be determined by the Gas Chromatography unless these acids are converted into Methyl Esters (Fatty Acid Methyl Ester). This video represents a detailed procedure of the sample preparation (or preparation of Fatty Acid Methyl Ester) for the analysis of Fatty Acid Composition of an oil or fat sample by acid-catalyzed esterification. __________________________________________________________________________________________
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Пікірлер: 138

@loveforyou201008 10 ай бұрын

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

@itizwhatitiz9560 Ай бұрын

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

@rushikeshfunde7531 Жыл бұрын

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

@MicroChemsExperiments Жыл бұрын

Good to know. Thanks for staying with us.

@pushtibhai3857 Жыл бұрын

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

@jualitafaradhaachnasya9463 Жыл бұрын

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!

@che5764 7 ай бұрын

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

@abigailabu9299 Ай бұрын

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

@MuhammadIbrahim-ty9bd 2 жыл бұрын

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

@MicroChemsExperiments 2 жыл бұрын

Noted. Shortly

@akankshasharma3755 2 жыл бұрын

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

@MicroChemsExperiments Жыл бұрын

Well done

@alperen1062 10 ай бұрын

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

@MicroChemsExperiments 10 ай бұрын

You can store for 1 month

@bijaychhetriconceptofzoolo1886 2 ай бұрын

Which article you are following to explain the GC process?

@md.riadulhoqehimal616 Жыл бұрын

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

@MicroChemsExperiments Жыл бұрын

Will upload soon

@md.riadulhoqehimal616 Жыл бұрын

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

@sushmithaj4099 2 жыл бұрын

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 2 жыл бұрын

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

@sushmithaj4099 2 жыл бұрын

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

@MicroChemsExperiments 2 жыл бұрын

@@sushmithaj4099 Okay

@crazyash1000 Жыл бұрын

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 Жыл бұрын

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.

@MrLongKR 8 ай бұрын

Hi. Which method is used for this experiment?

@rohitrajput8241 2 жыл бұрын

Please learn the further stem Sample inducting in GLC Instrument

@MicroChemsExperiments 2 жыл бұрын

@MrLongKR 5 ай бұрын

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

@MicroChemsExperiments 5 ай бұрын

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 Жыл бұрын

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

@MicroChemsExperiments Жыл бұрын

No, you can't

@deepasamnani7465 5 ай бұрын

I want a validation procedure for this method.

@nivedhitharamesh5936 2 жыл бұрын

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

@MicroChemsExperiments 2 жыл бұрын

Use micropipette or auto-sampler

@pushtibhai3857 Жыл бұрын

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 Жыл бұрын

Set condenser properly with cold water

@epalanisamiepalanisami6863 Жыл бұрын

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 Жыл бұрын

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

@epalanisamiepalanisami6863 Жыл бұрын

Thank you very much

@MicroChemsExperiments Жыл бұрын

@@epalanisamiepalanisami6863 You are welcome

@Eazycharts 2 ай бұрын

Why we add methanolic HCl?

@julietorsinime Жыл бұрын

thank you good video, injection in GC system analysis

@MicroChemsExperiments Жыл бұрын

Will upload soon

@shahriarislam1733 10 ай бұрын

which method is this?

@chamarakumara9222 2 жыл бұрын

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

@MicroChemsExperiments 2 жыл бұрын

Sure. Soon

@HishaniPrashar 10 ай бұрын

it's 2 nd part is uploaded

@rajanpatel3228 2 жыл бұрын

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

@MicroChemsExperiments 2 жыл бұрын

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

@kembarasembang497 2 жыл бұрын

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

@MicroChemsExperiments 2 жыл бұрын

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

@kembarasembang497 2 жыл бұрын

@@MicroChemsExperiments thank you so much sir. 😍

@ovaneschakoyan6584 2 жыл бұрын

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

@MicroChemsExperiments 2 жыл бұрын

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.

@ak-yh1jx Жыл бұрын

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

@MicroChemsExperiments Жыл бұрын

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

@epalanisamiepalanisami6863 Жыл бұрын

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

@MicroChemsExperiments Жыл бұрын

Methanolic BF3

@epalanisamiepalanisami6863 Жыл бұрын

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

@firdauskhan6154 2 жыл бұрын

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

@MicroChemsExperiments 2 жыл бұрын

Analyse as fast as possible (within 24h)

@godagamagds3820 Жыл бұрын

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

@MicroChemsExperiments Жыл бұрын

No. Methanol will not be appropriate.

@godagamagds3820 Жыл бұрын

@@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 Жыл бұрын

@@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.

@mariamshakeel468 Жыл бұрын

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

@MicroChemsExperiments Жыл бұрын

Try to find from journals

@MrLongKR Жыл бұрын

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

@MicroChemsExperiments Жыл бұрын

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

@shafiqahnabila5769 Жыл бұрын

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

@zeinasul Жыл бұрын

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

@MicroChemsExperiments Жыл бұрын

Not possible. You need to use GC

@23darshana Жыл бұрын

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

@MicroChemsExperiments Жыл бұрын

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

@23darshana Жыл бұрын

@@MicroChemsExperiments ok. Thanks.

@23darshana Жыл бұрын

can i get any reference to this method if possible?

@MicroChemsExperiments Жыл бұрын

@@23darshana no. Its a lab validated method

@akankshasharma3755 2 жыл бұрын

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 Жыл бұрын

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

@pushtibhai3857 Жыл бұрын

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

@MicroChemsExperiments Жыл бұрын

Concentration can be known by analyzing in GC

@dipalimalvekar2472 2 жыл бұрын

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

@MicroChemsExperiments 2 жыл бұрын

Same procedure

@izzatishrrrr 9 ай бұрын

what is the use of anhydrous sodium sulphate?

@MicroChemsExperiments 9 ай бұрын

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

@yahitajam7944 2 жыл бұрын

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

@MicroChemsExperiments 2 жыл бұрын

Why would you inject raw oil?

@yahitajam7944 2 жыл бұрын

@@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 2 жыл бұрын

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

@mirfazlerabbi9011 Жыл бұрын

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

@MicroChemsExperiments Жыл бұрын

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

@mirfazlerabbi9011 Жыл бұрын

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

@md.riadulhoqehimal616 Жыл бұрын

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

@MicroChemsExperiments Жыл бұрын

We will make video later

@epalanisamiepalanisami6863 2 жыл бұрын

Can we used sulphuric or nitric acid otherwise hcl

@MicroChemsExperiments 2 жыл бұрын

No. you must use HCl

@epalanisamiepalanisami6863 2 жыл бұрын

Thankyou very much . your experiments more useful

@DurayStuffCrafts 2 жыл бұрын

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

@MicroChemsExperiments 2 жыл бұрын

To absorb water/moisture from the extract

@vikashganvit3548 2 жыл бұрын

Hello ma'am Which colume your use for analysis ?

@MicroChemsExperiments 2 жыл бұрын

TG Polar

@vikashganvit3548 2 жыл бұрын

@@MicroChemsExperiments thank u ma'am

@MicroChemsExperiments 2 жыл бұрын

@@vikashganvit3548 you are welcome

@vikashganvit3548 2 жыл бұрын

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

@MicroChemsExperiments 2 жыл бұрын

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

@ovaneschakoyan7861 3 жыл бұрын

extraction time is it not too much?

@MicroChemsExperiments 3 жыл бұрын

Extraction should be faster

@dhruvamulani3084 8 ай бұрын

Can you share sample prepration reference name?

@MicroChemsExperiments 8 ай бұрын

It is a Laboratory developed in-house method

@tallapellyvikas6097 9 ай бұрын

Can you cite the reference of this derivatization procedure

@MicroChemsExperiments 9 ай бұрын

It is a inhouse laboratory developed method

@tallapellyvikas6097 8 ай бұрын

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

@aswinkumar8016 2 жыл бұрын

Part 2 ????

@MicroChemsExperiments 2 жыл бұрын

Please wait for the part-2

@aswinkumar8016 2 жыл бұрын

@@MicroChemsExperiments okay...

@afeeqafnani9690 Жыл бұрын

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

@MicroChemsExperiments Жыл бұрын

Soon

@MuhammadIbrahim-ty9bd 2 жыл бұрын

Part.2

@MicroChemsExperiments 2 жыл бұрын

Will upload soon

@aswinkumar8016 3 жыл бұрын

Derivatization

@MicroChemsExperiments 3 жыл бұрын

Yes. You are right

@aswinkumar8016 3 жыл бұрын

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

@MicroChemsExperiments 3 жыл бұрын

@@aswinkumar8016 Ok. We will make video later.

@aswinkumar8016 3 жыл бұрын

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

@MicroChemsExperiments 3 жыл бұрын

@@aswinkumar8016You are most welcome

@user-ci8sg4mz5k 11 ай бұрын

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

@MicroChemsExperiments 11 ай бұрын

Yes. You can