Some amino acids, serine, threonine, tryptophan, have double derivatives after derivatized by MTBSTFA (with GC MS), To get a good standard curve and quantification result, how to calculate peak area?
This has nothing to do with peak area. Integrate your peaks normally. This is all about good quantitative analysis, which means that you have to carry your amino acid standards (your calibration curve) through the derivatization process. You make your curve from underivatized amino acids and then carry the entire batch (curve, blanks, spikes, blank spikes and samples) through the derivatization procedure and then through the GC-MS analysis.
This is a really hard way to do amino acid analysis. Have you looked at CE or LC-MS?
Dear Mark, Sorry, I don't understandard"make your curve from underivatized amino acids and then carry the entire batch (curve, blanks, spikes, blank spikes and samples) through the derivatization procedure and then through the GC-MS analysis. "Is this the derivatization process? And I want to know how to quantification double derivatives, can you give some suggestions about this?
Dear Wahab, thank you for your answer. I have tried many times to miniminze the double derivatives since it cannot be avoided totally. Now I am afriad it would influence my quantification result. To make standard curve, I am not sure the peak area should contain 2 peaks or make 2 standard curve for each peak. For sum of peak area, the R2 is not good.
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