Miles Michael V, Tang Peter H, Miles Lili, Steele Paul E, Moye Mark J, Horn Paul S
Department of Pediatrics and Division of Pediatric Neurology, Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH 45229-3039, USA.
Biomed Chromatogr. 2008 Dec;22(12):1403-8. doi: 10.1002/bmc.1072.
Previous studies have indicated that analysis of coenzyme Q10 (CoQ10) in platelets may be clinically useful. The study objectives are to describe, validate and provide application of an HPLC-EC method for platelet CoQ10 analysis. This method analyzes oxidized (ubiquinone-10) and reduced (ubiquinol-10) forms of CoQ10 using two separate injections with the electrochemical analytical cell set at neutral and oxidizing potentials. Results showed that chromatograms were free of interfering peaks. Calibration curves were constructed over a concentration range 116-2317 nmol/L (r(2) = 0.99). The extraction recovery was >95%. The within-run precision CV% was < or =4.2%, and the day-to-day precision was < or =9.9%. Platelets were isolated by differential centrifugation, and frozen at -70 degrees C until analysis. The application of the method was used to compare accumulation of CoQ10 in platelets vs plasma in eight adult volunteers during a 28 day supplementation period (5 mg/kg/day of ubiquinol-10). Mean platelet total CoQ10 was 164 pmol/10(9) cells, and ubiquinol-10:total CoQ10 ratio was 0.56. During supplementation platelet CoQ10 levels were more consistent and predictable than plasma CoQ10 levels. The results indicate that this validated method for platelet ubiquinol-10 and ubiquinone-10 analysis is acceptable for use in the clinical laboratory, and that platelet CoQ10 may have important advantages over plasma during CoQ10 supplementation.
以往研究表明，分析血小板中的辅酶Q10（CoQ10）可能具有临床应用价值。本研究的目的是描述、验证并提供一种用于分析血小板CoQ10的高效液相色谱-电化学法（HPLC-EC）及其应用。该方法使用电化学分析池在中性和氧化电位下进行两次单独进样，分析CoQ10的氧化形式（泛醌-10）和还原形式（泛醇-10）。结果显示色谱图无干扰峰。在116 - 2317 nmol/L的浓度范围内构建校准曲线（r(2) = 0.99）。提取回收率>95%。批内精密度CV%≤4.2%，日间精密度≤9.9%。通过差速离心分离血小板，并在-70℃冷冻直至分析。该方法用于比较8名成年志愿者在28天补充期（5 mg/kg/天的泛醇-10）内血小板与血浆中CoQ10的积累情况。血小板中CoQ10的平均总量为164 pmol/10(9)个细胞，泛醇-10与总CoQ10的比率为0.56。在补充期间，血小板CoQ10水平比血浆CoQ10水平更一致且更可预测。结果表明，这种经过验证的用于分析血小板泛醇-10和泛醌-10的方法可用于临床实验室，并且在CoQ10补充期间，血小板CoQ10可能比血浆具有重要优势。
