INSULIN DEGRADATION INVIVO - A HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC ANALYSIS

被引:8
作者
BENZI, L [1 ]
CECCHETTI, P [1 ]
CICCARONE, AM [1 ]
DICIANNI, G [1 ]
IOZZI, LC [1 ]
CARICATO, F [1 ]
NAVALESI, R [1 ]
机构
[1] CNR,IST FISIOL CLIN,I-56100 PISA,ITALY
来源
JOURNAL OF CHROMATOGRAPHY-BIOMEDICAL APPLICATIONS | 1990年 / 534卷
关键词
D O I
10.1016/S0378-4347(00)82146-5
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
The metabolism of insulin in vivo was investigated using an isocratic reversed-phase high-performance liquid chromatographic (RP-HPLC) method. After intravenous injection of A14-[I-125]insulin into normals, eight labelled insulin derivatives were found in plasma (peaks 1-8). Two of them (peaks 1 and 7) showed an elution pattern identical with those of reference [I-125]monoiodotyrosine and intact A14-[I-125]insulin, respectively. Of the other six peaks, five (2-6) eluted before and one (peak 8) after insulin. This pattern was highly reproducible in terms of capacity factors and peak heights. Radioactivity separated by RP-HPLC was further characterized for its trichloroacetic acid precipitability and immunoprecipitability. Fractions corresponding to peaks 4-6 and 8, which showed an immunoprecipitability higher than 50%, were pooled in order to obtain sufficient radioactivity and were found to be insulin separated by Sephadex G-50 chromatography, containing in its structure, after sulphitolysis, intact A-chain and to be partially rebindable to monocyte insulin receptors. These data demonstrate that in blood, products of insulin metabolism circulate which retain a part of the immunological and biological properties of the hormone. These products are clearly separated from one another and from intact insulin by RP-HPLC, suggesting that the appropriate use of this technique may allow a further and more accurate qualitative and quantitative characterization of in vivo insulin metabolism in physiological and pathological conditions.
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页码:37 / 46
页数:10
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