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首页> 外文期刊>Analytical chemistry >Determination of Selenomethionine and Selenocysteine in Human Serum Using Speciated Isotope Dilution-Capillary HPLC-Inductively Coupled Plasma Collision Cell Mass Spectrometry
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Determination of Selenomethionine and Selenocysteine in Human Serum Using Speciated Isotope Dilution-Capillary HPLC-Inductively Coupled Plasma Collision Cell Mass Spectrometry

机译：同位素稀释-毛细管液相色谱-电感耦合等离子体碰撞细胞质谱法测定人血清中的硒代蛋氨酸和硒代半胱氨酸

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摘要

A method for the accurate determination of selenoamino acids in human serum by HPLC-ICPMS was developed using the speC_(16)s-specific isotope dilution analysis principle.A serum sample was enzymatically digested with a mixture of lipase and protease after derivatization of the selenocysteine residues with iodoacetamide.The selenoamino acid fraction was isolated by size exclusion LC followed by the separation of selenomethionine and the carboxymediylated selenocysteine by capillary HPLC.The isotope-specific determination of ~(77)Se and ~(80)Se was achieved on-line by ICP collision cell MS allowing the removal of polyatomic interferences.Quantification was carried out by isotope dilution using a ~(77)Se-labeled selenomethionine spike and the determination of the ~(77)Se/~(80)Se ratio in the cHPLC selenomethionine peak.The accurately determined selenomethionine was used as an internal standard for the selenocysteine determination from the same chromatogram.The modification of the previously developed cHPLC-ICPMS interface allowed the reduction of the absolute detection limits twice(down to the 75-fg level),which resulted in the lowest ever reported procedural detection limits(below 0.5 ng g~(-1)for a 450-mg serum sample).The precision was less than 5% RSD.The method was validated by the mass balance of selenium(amino acid incorporated vs total).
机译：建立了一种基于speC_（16）s特异性同位素稀释分析原理的高效液相色谱-ICPMS测定人血清中硒氨基酸的方法，将血清中的样品经脂肪酶和蛋白酶的混合物酶促消化后，将硒代半胱氨酸衍生化用碘色谱法分离硒氨基酸部分，然后通过毛细管HPLC分离硒代蛋氨酸和羧甲基化硒代半胱氨酸，通过同位素在线测定〜（77）Se和〜（80）Se通过ICP碰撞池MS消除多原子干扰。使用〜（77）Se标记的硒代蛋氨酸尖峰通过同位素稀释进行定量，并在cHPLC中测定〜（77）Se /〜（80）Se比硒代蛋氨酸峰。准确测定的硒代蛋氨酸被用作同一色谱图中硒代半胱氨酸测定的内标。先进的cHPLC-ICPMS接口使绝对检测限降低了两倍（降低至75-fg水平），从而实现了有史以来最低的程序检测限（450 mg低于0.5 ng g〜（-1））精密度低于RSD的5％。该方法通过硒的质量平衡（掺入的氨基酸与总量的硒）进行验证。

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