血浆中萘丁美酮活性代谢物的反相高效液相色谱法测定

Determination of 6-methoxy 2-naphthylacetic Acid,a Major Metabolite of Nabumetone,in Human Plasma by HPLC

为研究萘丁美酮的相对生物利用度，建立了用反相高效液相色谱法测定血浆中萘丁美酮的活性代谢物６－甲氧基－２－萘乙酸（ＭＮＡ）浓度的方法。实验结果表明，以萘普生作内标，甲醇∶０．０２ｍ ｏｌ／Ｌ醋酸盐缓冲液（ｐＨ３．０）为７４∶２６ 作流动相，用ＹＷＧ－Ｃ１８ 柱有良好的分离效果；血浆样品在ｐＨ３．０醋酸盐缓冲液存在下，用二氯甲烷萃取，回收率高，在２７０ｎｍ 波长下检测杂质峰少。该法标准曲线在０．５～６４ｍ ｇ／Ｌ范围内有良好线性，最低检测浓度为０．０２ｍ ｇ／Ｌ，萃取回收率为８８％ ～９４％ ，方法回收率为９６％ ～１０２％ ，日内相对标准差小于３．５％ ，日间相对标准差小于５％ 。具有快速简便，灵敏准确等特点。

This paper reports a sensitive and rapid method for determining 6 methoxy 2 naphthylacetic acid(MNA), a major metabolite of nabumetone in human plasma using naproxen as the internal standard. High performance liquid chromatograph model 680(Waters, USA) with a variable wavelength UV detector and reversed phase YWG C 18 column(10μm, 250×4.6mm) was used. After the addition of acetate buffer(pH3.0), the plasma sample was extracted with methylene chloride. The mobile phase of methanol pH3.0, 0.02 mol/L acetate buffer(74∶26) was pumped at 1.0 ml/min through the column. The detector at 0.01AUFS was set at 270nm. The retention times for MNA and naproxen were 3.98min and 4.73 min respectively. Standard curve was linear in the concentration range of 0.5 to 64 mg/L. The detection limit in serum was 0 02 mg/L. Extraction recovery was 88% 94%; method recovery 96% 102%; withinday RSD less than 3.5%; inter day RSD less than 5%.