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为何应摒弃通过蛋白质沉淀和色谱法测定血清胱氨酸的方法。

Why the assay of serum cystine by protein precipitation and chromatography should be abandoned.

作者信息

Cooper J D, Turnell D C, Green B, Wright D J, Coombes E J

机构信息

Biochemistry Department, Coventry and Warwickshire Hospital, Wiltshire, UK.

出版信息

Ann Clin Biochem. 1988 Sep;25 ( Pt 5):577-82. doi: 10.1177/000456328802500516.

Abstract

The higher bias of serum cystine estimations by a HPLC method compared with those by ion exchange techniques is shown to be largely due to differences in the sample preparation procedures of the two techniques. The ion exchange methods utilised sulphosalicylic acid serum protein precipitation and post-column ninhydrin derivatisation of cystine, whilst the high pressure liquid chromatography technique employed automated dialysis for removal of proteins and pre-column ortho-phthalaldehyde derivatisation of cystine after its conversion to cysteine and then to S-carboxymethylcysteine. Examination of these procedures showed that whilst the high pressure liquid chromatographic method accurately estimates total serum cystine and cysteine, many factors affect the precision and accuracy of serum cystine estimations using the ion exchange techniques. In particular, serum protein precipitation techniques that are currently employed for the preparation of samples for cystine analysis by ion exchange chromatography should be abandoned.

摘要

与离子交换技术相比,高效液相色谱法测定血清胱氨酸时偏差更高,这在很大程度上归因于两种技术样品制备程序的差异。离子交换方法利用磺基水杨酸沉淀血清蛋白并对胱氨酸进行柱后茚三酮衍生化,而高压液相色谱技术采用自动透析去除蛋白质,并在胱氨酸转化为半胱氨酸后再转化为S - 羧甲基半胱氨酸后对其进行柱前邻苯二甲醛衍生化。对这些程序的检查表明,虽然高压液相色谱法能准确测定血清总胱氨酸和半胱氨酸,但许多因素会影响使用离子交换技术测定血清胱氨酸的精密度和准确性。特别是,目前用于通过离子交换色谱法分析胱氨酸的样品制备的血清蛋白沉淀技术应该被摒弃。

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