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A simple steam distillation method of estimating β-hydroxy-α-amino acids

✍ Scribed by D.R. Keeney; J.M. Bremner

Publisher
Elsevier Science
Year
1967
Tongue
English
Weight
684 KB
Volume
18
Category
Article
ISSN
0003-2697

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✦ Synopsis

Nicolet and Shinn (1, 2) showed that P-hydroxy-a-amino acids react with periodate in alkaline medium according to the following equation: RCH(OH)CH(NH&R' + Na101 = RCHO + R'CHO + NI& + NaIOs This reaction has formed the basis for a number of methods of estimating serine, threonine, and other amino acids containing -CH(OH) CH (NH,)-groups. Methods involving determination of the ammonia liberated by this reaction have the attraction that ammonia can be determined by relatively simple procedures, and several methods in which the ammonia released by periodate is determined by aeration or microdiffusion techniques have been described (3-11). Van Slyke, Hiller, and MacFadyen

(4) made a detailed study of the conditions required for quantitative determination of hydroxyamino acids in protein hydrolyzates from the ammonia liberated by alkaline periodate and developed a procedure involving treatment of the hydrolyzate sample with glycine, sodium hydroxide, periodic acid, and potassium carbonate solution, and subsequent determination of the ammonia liberated by aeration of the treated sample at room temperature for 30 min. Since this procedure recovers ammonium present in the sample before addition of periodate, an additional analysis involving determination of the ammonia liberated by aeration of a duplicate sample with potassium carbonate solution is required to allow for this ammonium.

The periodate-ammonia method of determining p-hydroxy-a-amino acids described here developed from an attempt to simplify the method of Van Slyke, Hiller, and MacFadyen by use of steam distillation instead of aeration for estimation of the ammonia released by periodate. It involves steam distillation with phosphate-borate buffer (pH 11.2) for 4 min before and after treatments with periodic acid and sodium arsenite,

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