How would you detect lipids with GC? 8
Which FAME standards and internal standards would be best for calibration? It's my understanding that I would run different concentrations of FAME standards + 2 internal standards with GC. Then I can correlate the area under the chromatograph with concentration in a calibration curve. For each sample I run after, I spike it with the internal standards at a known concentration. Is this correct?
Fri December 03 2010 06:33:39 AM by Green123
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For calibration of GC: When you have FAME standard mixture for example from sigma-aldrich, The weight percentage of each component is indicated. Each ampule contains 10 mg/mL of the FAME reference standard mix in methylene chloride (LINK). You have to run GC injecting that FAME mixture. From the resulting chromatogram you can detect the retention time of each fatty acid and compare the percent of each one with that of Sigma.
To detect the concentration of new samples: you have to add internal standard (ex. 17:0 or 19:0...) in a certain weight (ex. 8 Microgram). It is important to be sure that your sample doesn't contain this standard. From the peak area of fatty acids, you can calculate the weight of each one comparing with the standard's peak area.
Best Wishes,
Abomohra
Good dr abdelfatah but there are some suggestions from me when you detect lipid by GC or HPLC(the best) some troubleshoting will you face retention time from standard and sample will be shifting from 0.1-0.5 min but allowed from 0.1-0.3 min and actually ocurred so we solving this problem by adding standard to sample and calculate the difference between them and it's the best method as we know there are many peaks appear in our chromatogrames and also each time inject standard as some systems have data library And dr if you mean GC- mass spectra I am with you.
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