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Assay for β-ureidopropionase by high-performance liquid chromatography

✍ Scribed by Günther Waldmann; Beate Podschun

Publisher
Elsevier Science
Year
1990
Tongue
English
Weight
376 KB
Volume
188
Category
Article
ISSN
0003-2697

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✦ Synopsis

A sensitive assay for beta-ureidopropionase based on derivatization of the reaction product beta-alanine with phenylisothiocyanate has been developed. Purification of the resulting phenylthiocarbamoyl-beta-alanine is achieved on a LiChrospher 100 C18 reversed-phase high-performance liquid chromatography column using an isocratic elution system. Phenylthiocarbamoyl-beta-alanine is detected by its absorbance at 245 nm and quantitated by automatic peak integration referring to a calibration curve. This technique offers a high degree of sensitivity as beta-alanine quantities in the picomole range can be identified. N-Carbamoyl-beta-alanine, the natural substrate of beta-ureidopropionase, does not interfere with the described assay system. The enzymatic reaction is linear for an incubation time of 45 min with enzyme concentrations of 3.2 micrograms/ml.

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