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采用柱后串联库仑法和紫外检测的液相色谱法测定生物样品中的辅酶Q10、α-生育酚和胆固醇。

Determination of coenzyme Q10, alpha-tocopherol and cholesterol in biological samples by coupled-column liquid chromatography with coulometric and ultraviolet detection.

作者信息

Edlund P O

机构信息

Bioanalytical Section, Research and Development, ACO Läkemedel AB, Solna, Sweden.

出版信息

J Chromatogr. 1988 Mar 4;425(1):87-97. doi: 10.1016/0378-4347(88)80009-4.

DOI:10.1016/0378-4347(88)80009-4
PMID:3360880
Abstract

Coenzyme (Co) Q10, Co Q10H2, alpha-tocopherol and cholesterol were dissociated from lipoproteins in plasma by treatment with 1-propanol. The supernatant obtained was injected directly for determination of Co Q10 and Co Q10H2. Precolumn reduction with borohydride was used for determination of total Co Q10 simultaneously with alpha-tocopherol and cholesterol. Total Co Q10 in freeze-dried myocardial biopsies was determined after extraction with 1-propanol and oxidation of Co Q10H2 with ferric chloride. The chromatographic system comprised two reversed-phase columns and a three-electrode coulometric detector and a UV detector coupled in series. A pre-fractionation on the first column protected the coulometric detector from contamination and reduced the time for analysis by eliminating strongly retained solutes. The coulometric electrodes were operated in the oxidation-reduction-oxidation mode, and the last electrode was used for detection of alpha-tocopherol, Co Q10 and Co Q10H2, while cholesterol was detected by UV at 215 nm. The fast isolation procedure made it possible to determine the reduced and oxidized forms of Co Q10 in plasma. Quantitative recoveries were obtained for all the analytes studied and normal levels were determined with a coefficient of variation of 2-3%.

摘要

通过用1-丙醇处理,从血浆脂蛋白中分离出辅酶(Co)Q10、Co Q10H2、α-生育酚和胆固醇。将获得的上清液直接进样以测定Co Q10和Co Q10H2。用硼氢化物进行柱前还原,同时测定总Co Q10以及α-生育酚和胆固醇。冻干心肌活检组织中的总Co Q10在用1-丙醇提取并将Co Q10H2用氯化铁氧化后进行测定。色谱系统由两根反相柱以及一个三电极库仑检测器和一个串联的紫外检测器组成。在第一根柱上进行预分离可保护库仑检测器不受污染,并通过去除强保留溶质来减少分析时间。库仑电极以氧化-还原-氧化模式运行,最后一个电极用于检测α-生育酚、Co Q10和Co Q10H2,而胆固醇则在215nm处通过紫外检测。这种快速分离程序使得能够测定血浆中Co Q10的还原形式和氧化形式。对所有研究的分析物均获得了定量回收率,测定的正常水平变异系数为2%-3%。

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