HPLC法测定大鼠血浆中乙醛的含量

目的: 建立检测大鼠血浆中乙醛(ACH)浓度的HPLC方法。 方法: 选择正丁醛-2,4-二硝基苯肼(DNPH)为内标,采用ZORBAX Eclipse XDB-C₁₈(250 mm×4.6 mm,5 μm)色谱柱,以甲醇-乙腈-水(25:35:40,v/v/v)为流动相,流速1.0 mL·min^-1,在紫外波长为360 nm处检测。血浆样品需经乙腈沉淀蛋白,离心后取上清与DNPH发生衍生化反应,正己烷-乙酸乙酯(5:1,v/v)萃取衍生化产物ACH-DNPH,进行高效液相色谱测定。 结果: 乙醛在0.05~10.00 μg·mL^-1浓度范围内线性关系良好(r=0.9999),检测限为0.01 μg·mL^-1,定量限为0.05 μg·mL^-1;高、中、低3个浓度的相对回收率为102.7%~109.0%;日内RSD为1.6%~4.3%,日间RSD为3.3%~6.3%。 结论: 本法可用于大鼠体内乙醇代谢动力学研究。

Objective: To establish a method for determination of acetaldehyde(ACH) in rat plasma by HPLC. Methods: With butyl aldehyde-2,4-dinitrophenylhydrazine(BUH-DNPH) as the internal standard,ACH-DNPH was detected by UV detector at the wavelength of 360 nm with the ZORBAX Eclipse XDB-C₁₈ (250 mm×4.6 mm,5 μm) column.The mobile phase was methanol-acetonitrile-water (25:35:40,v/v/v) and the flow rate was 1.0 mL·min^-1.The plasma samples were treated with acetonitrile to precipitate proteins.After centrifugation,the supernatant was reacted with DNPH reagent.The derived acetaldehyde was then isolated by n-hexane-acetidin(5:1, v/v) and quantitated by high performance liquid chromatography. Results: The assay linearity of ACH was confirmed over the range of 0.05-10.00 μg·mL^-1 (r=0.9999).The limit of detection was 0.01 μg·mL^-1 and the limit of quantitation was 0.05 μg·mL^-1.The relative recoveries at high,medium and low concentrations were within 102.7%-109.0%,and the RSD of intra-day and inter-day assays were 1.6%-4.3% and 3.3%-6.3%,respectively. Conclusion: The method is suitable for the pharmacokinetic research of alcohol in rats.

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