4/22/2018 Can you calculate percentage composition of fatty acids from...

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Question Asked 4 years ago
Stephen Mackay
4.53 · University of Alabama at Birmingham
Can you calculate percentage composition of fatty acids from a sample without
a calibrated external standard?
I ran GCMS samples of microalgae in the past to calculate FAME composition. I
ran it with 2 internal samples C13:0 before extraction and C19:0 post extraction.
The data I obtained only gave me peak areas. (The only standard available was
the library for peak identi cation). As I was in a visiting lab and had to run the
samples alone as the external standards were not available to me. Is it possible
to at least calculate relative composition from this data based solely on peak
values, normalized by the internal standards? I was told I could but now I'm not
con dent it can be done accurately.

Microalgae Biodiesel Microalgae Molecular Biological Techniques Microbiology

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Popular Answers (1)

Yannis Karamanos 4 years ago
Université d'Artois

Ideally one should run a standard FAME mixture together with the two internal
standards (e.g. C13:0 and C19:0) and from that calculate a conversion coef cient
for each individual FAME (that is the ratio area of a given FAME to the area of the
internal standard (you can plot it for each internal standard). Then you should inject
the "unknown" sample without the internal standards to be sure that no C13:0 or
C19:0 occurs in the sample naturally. After this control you can use the internal
standards without any concern. Now, using the conversion coef cients you will
obtain accurate calculations for the relative composition.

If you do not have the possibility to use a standard FAME mixture you should be
aware that differences could occur in the response (area) of individual fatty acids.

3 Recommendations
Question followers (13) See all
All Answers (7)
Yannis Karamanos 4 years ago
Université d'Artois

Ideally one should run a standard FAME mixture together with the two internal Similar Questions
standards (e.g. C13:0 and C19:0) and from that calculate a conversion coef cient
for each individual FAME (that is the ratio area of a given FAME to the area of the How do you calculate Fatty acid methyl esters
internal standard (you can plot it for each internal standard). Then you should inject percentage from a GC graph?
the "unknown" sample without the internal standards to be sure that no C13:0 or Give detailed explanation about calculation of FAME %.
C19:0 occurs in the sample naturally. After this control you can use the internal 5 answers added
standards without any concern. Now, using the conversion coef cients you will
obtain accurate calculations for the relative composition.
Can anyone suggest how to calculate the concentration
If you do not have the possibility to use a standard FAME mixture you should be of the compound through peak area and RT using
aware that differences could occur in the response (area) of individual fatty acids. GCMS for FAME?
We are working on Microalgae biodiesel. For  FAME analysis
 GC MS was carried out without standard run and Peak area
3 Recommendations
and RT is given in data. So...
Jai Ghosh 4 years ago
10 answers added
Added an answer

The peak areas will tell you the content of each of the FAME which if you add up How to express/calculate fatty acids represented as
should give you % composition. g/100g fatty acids (relative proportion) in mole percent
(mol%) ?
Luis M. Rodríguez-Alcalá 4 years ago Analysis of fatty acid methyl esters (FAME) has been done by
Universidade Católica Portuguesa GC. How do we calculate FAME in mole percent using the MW
of the fatty acids?
Hi Stephen, 5 answers added

Yannis has answered you very nicely however I would like to add some comments:
First of all you should run a FAME mix to calculate response factors (Yannis´ What is the most reliable method to do quantitative
conversion coef cient). With this you calibrate your MS response. This will give you analysis of fatty acid methyl esters (FAMEs) in
a factor to correct your areas in order the MS is giving different signals to each fatty ecological studies?
acid (and it does!). Only with this correction you may express your results as "area The final output is the profile of all FAs in the analysed
percent". Ok, them you check if your samples has your C13 and your C19. If not or sample. So should I choose normalization, internal or external
the percent are low (<0.5%) you can use your standards. You can also use the FAME standardization?
mix to run calibration curves for every fatty acid and it is ok, but you need to make 20 answers added
several runs with different concentrations. So, with the response factors to correct
the peaks areas and the internal standards you can calculate accurately your
How can I calculate the concentration from a gc/ms
contents in concentration (i.e. mg FA/g algae DM) which is for me the right way to
chromatogram which is having retention time and peak
do this.
area only?
How can I know the area of ISI?  Was used an internal
Let´s imagine you have not a FAME mix standard. Well, I imagine that in the lab may
standard which has c20-c 40 alkane standard. How could I
have a sh or high omega 3 oil. If you have a certi ed composition you can used in
found the area of my IS as it is...
the same way that the FAME mix but you will have to prepare the FAME.

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4/22/2018 Can you calculate percentage composition of fatty acids from...
Good luck and keep us updated!. 24 answers added

Stephen Mackay 4 years ago How can I calculate % of gc-yield or conversion? is

University of Alabama at Birmingham there any use of internal standard?
please inform in details as i am a beginner in this field. we
Thanks for the responses. I did not mention that we had used a fame and bame have gc-FID facility 
library as to at least identify the respective peaks. I understood the calculation as
you have described above. Unfortunately I had run the experiments in a previous lab 9 answers added
and was not able to run standards and wont be able to run them again. Was
wondering if there was a way to express the data without those standards. How can I calculate the yield of the products from a GC
chromatogram which is having retention time and peak
But as I understand from your responses that is not possible to use the raw peak area only?
data in anyway. I had hoped i could do it like Jai had suggested but was not sure if Thanks in advance for your replies.
that is accurate.
15 answers added
Thanks again for the clear responses.
How to calculate FAME conversion yield from GC peak
Luis M. Rodríguez-Alcalá 4 years ago area after enzymatic transesteri cation?
Universidade Católica Portuguesa I converted salmon oil into FAME using lipase. I used BSFTA
and tetrahydrofuran to derivatize the sample and injected in
Hi Stephen, GC for separation and...
5 answers added
Jai is right but the results would not be accurate.

Elena Koudouna 4 years ago Is there a conversion factor for the calculation of fatty
Doshisha University acid content of microalgae?
I am doing a research in microalgae and I did analysis for
Hi Stephen, lipid composition and fatty acid profile. To calculate the fatty
acid content (% DW),...
Like the people mentioned above ideally you should use calibrated external 3 answers added
standards otherwise you will only get the relative percentage concentration of fatty
acids
How can I calculate the conversion and the yield of
biodiesel from the GC graph?
Md. Washim Akram a month ago
How can I calculate the conversion and the yield of biodiesel
Independent Researcher
from the Gas Chromotography graph?

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