A rapid method for the extraction and determination of vitamin E metabolites in human urine.
A rapid method for the extraction and determination of vitamin E metabolites in human urine.
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一种快速提取并测定人尿液中维生素E代谢物的方法。
DOI：
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发表时间：
2000
影响因子：
6.5
通讯作者：
Regina Brigelius
中科院分区：
文献类型：
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作者：
John K. Lodge;M. Traber;A. Elsner;Regina Brigelius
建立了人尿中维生素E代谢物γ-和α-羧乙基羟色曼(γ-和α-CEHC)的直接提取和常规分析方法。可以使用相对较小的样品体积(5毫升)，在酶解结合形式和酸化之后，用二乙醚提取代谢物。将α-和伽马-CEHC的回收率与作为内标的曲洛克斯的回收率进行比较，并将其添加到未补充维生素E的志愿者的24小时尿液中。对不同的溶剂条件进行了初步试验，酸化和乙醚提取的回收率最高。经加入和从尿液中提取后，曲洛克斯、α-和γ-CEHC的回收率相似，因此曲洛克斯作为内标是可行的。用气相色谱和高效液相色谱电化学检测(ECD)对样品进行了分析。与GC或290 nm紫外检测的高效液相色谱相比，高效液相色谱-电子捕获检测器具有更高的选择性和灵敏度。高效液相-电子捕获检测器的检出限为10fmol，线性范围为40~200fmol(r(2)>；0.98)，对未添加补充剂的受试者24小时尿液检测效果最佳。样本间变异通常为2-5%。这种更高的灵敏度和选择性意味着即使在没有补充维生素E的受试者中也可以分析维生素E代谢物。也可以测量代谢物的非结合形式。通常，这些元素约占α-和伽马-CEHC总量的10%。该方法可用于尿液中维生素E代谢物的常规测定。所描述的新的提取和检测方法比以前的方法相对快速、省力且更具成本效益。
A method for the direct extraction and routine analysis of the vitamin E metabolites gamma- and alpha-carboxyethyl hydroxychroman (gamma- and alpha-CEHC) from human urine has been developed. A relatively small sample volume (5 ml) can be used and, after enzymatic hydrolysis of the conjugated forms and acidification, the metabolites are extracted with diethyl ether. Recovery of alpha- and gamma-CEHC was compared to that of trolox, used as an internal standard, added to 24-h urine collections from vitamin E-unsupplemented volunteers. Various solvent conditions were initially tested; acidification and ether extraction gave the highest recovery. It was found that after addition and extraction from urine, trolox, alpha- and gamma-CEHC are recovered to a similar extent, hence trolox is viable as an internal standard. The samples were analyzed by both GC and HPLC with electrochemical detection (ECD). HPLC-ECD was found to give higher selectivity and higher sensitivity compared to GC or HPLC with UV detection at 290 nm. The HPLC-ECD detection limit was 10 fmol, linearity (r(2) > 0.98) was achieved in the range of 40 to 200 fmol, which was found to be optimal for 24-h urines from unsupplemented subjects. Inter-sample variability was typically 2-5%. This greater sensitivity and selectivity means that vitamin E metabolites can be analyzed even in unsupplemented subjects. It is also possible to measure unconjugated forms of the metabolites. Typically these were found to represent approximately 10% of the total alpha- and gamma-CEHC. This method can be used routinely for the determination of vitamin E metabolites in urine. The new extraction and detection methods described are relatively quick, less laborious, and more cost-effective than previously available methods.
DOI：
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发表时间：
1997-08
期刊：
The Journal of pharmacology and experimental therapeutics
影响因子：
--
作者：
E. D. Murray;W. Wechter;D. Kantoci;W. Wang;T. Pham;D. Quiggle;K. Gibson;D. Leipold;B. Anner
通讯作者：
E. D. Murray;W. Wechter;D. Kantoci;W. Wang;T. Pham;D. Quiggle;K. Gibson;D. Leipold;B. Anner
同时测定组织生育酚、生育三烯酚、泛醇和泛醌。
影响因子：
6.5
作者：
Podda,M;Weber,C;Traber,MG;Packer,L
通讯作者：
Packer,L