Amino acid part analysis.

Ask: I have a protein want to do
amino acids
-part analysis, want to use 6MHCl treatment into a single amino acid after using high-efficiency liquid like
chromatography
to do, please ask how to do, thank you?
: This question is relatively large, ha ha, have done a little before, and the landlord to discuss it.
First of all, amino acid composition analysis workload is not small, and requires the operator to have a higher level of operation, if you just want to test it, it is better to spend some money sent out to measure, do their own sometimes in vain ah. Liquid chromatography can be done, but the testing agency has a dedicated amino acid analyzer (although the core is also an HPLC).
if the landlord insists on doing it himself, then I say something I knew before, that is, a rough one, because it's been more than two years. I hope I can help you. The experimental process of amino acid composition analysis (chromatography) is divided into three steps: hydrolysing, derivative and chromatography.
first of all, hydrolyzing, mainly acid and enzyme method, acid method will have a destructive effect on certain amino acids, and enzyme method, although mild, but sometimes hydrolyzed is not complete. Or the use of acid method more. The acid method can be added with 2% phenol as a protective agent to prevent the Met and Cys from being oxidized;
right, Cys is best to protect, otherwise quantitative not allowed, the method of protection can be alkylated, the specific process can check the literature. The whole process is best vacuumed and filled with nitrogen.
diffraction
bio-chemical
methods are many, with many OPA (with fluorescent detector) and PTC (with 254nm ultraviolet detection), both are derived before the column, that is, before hpLC derivative. The owner of this building can check the literature, I do not remember the specific operation. Basically those testing agencies are much more useful for these two derivatives.
color spectrum is simple, generally is a phase A sodium acetate, B phase methanol or acetylene. The two methods come out in a different order, and you can find a relevant document using a method to know the order. Then the integral analysis is on the line. However, the amino acid composition analysis error is not small, 5 to 10%, or even greater.
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