Hydrolysis

Hydrolysis is typically achieved by acid conditions. A standard procedure is hydrolysis with 6 M hydrochloric acid (24 hours, 110°C). These standard procedure is a compromise between time requirement and temperature. Sensitive amino acids (especially tryptophane and cysteine) will be partially destroyed. Gas phase hydrolysis and addition of other acids (e.g. propionic acid, TFA, methansulfonic acid) can be used to shorten the hydrolysis time and improve the yield of sensitive amino acids.

Table 1.

Catalog No.	Product Name
77249	Heptafluorobutyric acid
84429	Hydrochloric acid Solution, 6 M in water
45173	Endoproteinase Arg-C
45168	Endorpoteinase Asp-N
45169	Endoproteinase recombinant
45172	Endoproteinase Glu-C
45175	Endorpoteinase Lys-C
45167	Endoproteinase Pro C
27207	Chymotrypsin

Separation, detection and quantification

Hydrolysed samples(amino acids)are derivatized for sensitive detection,separated by HPLC. Whereas post-column derivatization was typical earlier, pre-column derivatization has gained importance and can be achieved by a broader range of derivatization reagents. The use of internal and external standards is crucial. A selection of well suited products is listed below(see Table 2).

Table 2.

Catalog No.	Product Name
88559	2,2'-Thiodiethanol
12715	Benzo[f]ninhydrin Monohydrate
21795	Carbazole-9-carbonyl chloride
23182	(+)-1-(9-Fluorenyl)ethyl chloroformate solution
23183	(-)-1-(9-Fluorenyl)ethyl chloroformate solution
23184	9-Fluorenylmethyl chloroformate
39065	4-Dimethylaminoazobenzene-4'-isothiocyanate
39068	4-Dimethylaminoazobenzene-4'-sulfonyl chloride
39220	5-(Dimethylamino)-1-naphthalene sulfonylchloride
63707	2-Mercaptoethane sulfonic acid solution
42095	N-a-2,4-Dinitro-5-fluorophenyl)-L-alaninamide
42100	N-a-2,4-Dinitro-5-fluorophenyl)-D-valinamide
42102	N-a-2,4-Dinitro-5-fluorophenyl)-L-valinamide
47614	Fluorescamine
72490	Ninhydrin
78779	Phenyl isothiocyanate
78781	Phenyl isothiocyanate
79760	Phthaldialdehyde
86729	2,3,4,6-Tetra-O-benzoyl-b-D-glucopyranosyl--isothiocyanate
92822	2,3,4,6-Tetra-O-benzoyl-b-D-glucopyranosyl isothiocyanat solution 1 M in water
89485	Titanium(III) chloride solution
87363	Tetrahydrofuran
90337	Triethylamine
33484	1,8-Diazafluoren-9-one
09418	Amino acids Standard Solution, 0.01 mM each
09421	Amino acids Standard Solution, 0.05 mM each
09423	Amino acids Standard Solution, 0.1mM each
09425	Amino acids Standard Solution, 0.5 mM each
09428	Amino acids Standard Solution, 1 mM each
15411	Boc-Cys-OH
58689	N-Isobutyryl-D-cysteine
58698	N-Isobutyryl-L-cysteine