一种测定大鼠血浆中生育酚的高效液相色谱新方法

S. Ghodbane, A. Lahbib, S. Ennaceur, M. Sakly, M. R. Driss
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引用次数: 3

摘要

目的:简化高效液相色谱法测定大鼠血浆中维生素E的样品处理方法,评价镉处理对血浆中维生素E含量的影响。方法:建立了用正己烷-二氯甲烷直接萃取法测定维生素E的高效液相色谱法。干燥的残渣在甲醇中再溶解。该方法采用Supelco Discovery®C18色谱柱,甲醇/水(95:5,v/v)为流动相。开发后,用荧光检测器对该方法进行了验证。为了研究外源性镉对血浆维生素E浓度的影响,采用体重150±20 g的雄性Wistar大鼠,分别口服15和30 mg/L镉(以CdCl 2形式)和维生素E (10 mg/Kg日粮)48 h和10 d。结果:该方法使样品处理更容易,特别是在处理大量样品时。事实证明,它具有选择性、线性性、准确性和精确性。镉中毒后血浆维生素E浓度显著下降。30 mg/L给药组在给药10 d后达到最低水平。结论:建立了一种快速反相高效液相色谱法测定血浆中维生素E含量的方法。该方法可用于常规检测,节省了溶剂和色谱时间。有趣的是,α -生育酚可能通过清除自由基在预防镉暴露引起的氧化应激中发挥重要作用。
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A new HPLC method for the determination of tocopherol in rat plasma
Objective: The objective of the present work is the simplification of sample treatment for vitamin E measurements in rat plasma by high-performance liquid chromatography (HPLC), then the evaluation of the effect of cadmium treatment on the vitamin E level. Methods: A HPLC method to determine vitamin E was developed with direct extraction with n-hexane-dichloromethane. The dry residue was redissolved in methanol. The method employs a Supelco Discovery ® C18 column and methanol/water (95:5, v/v) as the mobile phase. After being developed, the method was validated with a fluorescence detector. To examine the effects of exogenous cadmium on the plasma vitamin E concentrations, male Wistar rats (weighing 150 ± 20 g) were exposed for 48 h and 10 days to oral intake of 15 and 30 mg/L cadmium (as CdCl 2 ) and vitamin E (10 mg/Kg of diet) simultaneously. Results: This method makes sample treatment easier, especially when working with a large number of samples. It has proved to be selective, linear, accurate and precise. Intoxication with cadmium was followed by a significant decrease in plasma vitamin E concentrations. The lowest level was obtained in the 30 mg/L dosing group after 10 days of cadmium administration. Conclusion: We describe a rapid reversed-phase (RP) HPLC procedure for the determination of vitamin E in plasma. This method might be useful in routine assessment because it saves on solvents and chromatographic time. Interestingly, α -tocopherol may play an important role in preventing oxidative stress induced by cadmium exposure by scavenging free radicals.
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