A procedure suitable for a selective high-performance liquid chromatographic (HPLC) analysis of the imino acid hydroxyproline in the presence of a large excess of amino acids is proposed. To deaminate the amino acids, the well-known reaction with nitrous acid is exploited. The N-nitroso derivatives of imino acids obtained are extracted in ethyl acetate, denitrosated by hydrobromic acid, and treated with dansyl-chloride. The final HPLC separations are carried out on a reversed-phase column in a rapid isocratic run. The use of the cis isomer of hydroxyproline as an internal standard allows good reproducibility. As an application of the described method, the hydroxyproline content in samples containing collagen and an excess of bovine serum albumine (up to 20:1) is determined.

A new procedura for the specific high-performance liquid chromatographic determination of hydroxyproline / P.A. Biondi, L.M. Chiesa, M.R. Storelli, P. Renon. - In: JOURNAL OF CHROMATOGRAPHIC SCIENCE. - ISSN 0021-9665. - 35:11(1997), pp. 509-512.

A new procedura for the specific high-performance liquid chromatographic determination of hydroxyproline

P.A. Biondi
Primo
;
L.M. Chiesa
Secondo
;
P. Renon
Ultimo
1997

Abstract

A procedure suitable for a selective high-performance liquid chromatographic (HPLC) analysis of the imino acid hydroxyproline in the presence of a large excess of amino acids is proposed. To deaminate the amino acids, the well-known reaction with nitrous acid is exploited. The N-nitroso derivatives of imino acids obtained are extracted in ethyl acetate, denitrosated by hydrobromic acid, and treated with dansyl-chloride. The final HPLC separations are carried out on a reversed-phase column in a rapid isocratic run. The use of the cis isomer of hydroxyproline as an internal standard allows good reproducibility. As an application of the described method, the hydroxyproline content in samples containing collagen and an excess of bovine serum albumine (up to 20:1) is determined.
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2434/58352
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