Cham B E, Bochner F, Imhoff D M, Johns D, Rowland M
Clin Chem. 1980 Jan;26(1):111-4.
We have developed a specific and sensitive method for the determination of salicylic acid, salicyluric acid, and gentisic acid in urine. Any proteins present are precipitated with methyl cyanide. After centrifugation, an aliquot of the supernate is directly injected into an octadecyl silane reversed-phase chromatographic column, then eluted with a mixture of water, butanol, acetic acid, and sodium sulfate, and quantitated at 313 nm by ultraviolet detection according to peak-height ratios (with internal standard, o-methoxybenzoic acid) or peak heights (no internal standard). The method allows estimates within 25 min. Sensitivity was 0.2 mg/L for gentisic acid, and 0.5 mg/L for both salicyluric and salicylic acid (20-micro L injection volume); response was linear with concentration to at least 2.000 g/L for salicylic acid and metabolites. Analytical recovery of salicylic acid and metabolites from urine is complete. Intra-assay precision (coefficient of variation) is 5.52% at 7.5 mg/L for salicylic acid, 5.01% at 9.33 mg/L for salicyluric acid, and 3.07% at 7.96 mg/L for gentisic acid. Interassay precision is 7.32% at 7.51 mg/L for salicylic acid, 5.52% at 8.58 mg/L for salicyluric acid, and 3.97% at 8.32 mg/L for gentisic acid. We saw no significant interference in urine from patients being treated with various drugs other than aspirin.
我们已开发出一种用于测定尿液中水杨酸、水杨尿酸和龙胆酸的特异且灵敏的方法。存在的任何蛋白质都用乙腈沉淀。离心后，取一份上清液直接注入十八烷基硅烷反相色谱柱，然后用水、丁醇、乙酸和硫酸钠的混合物洗脱，并根据峰高比（以内标邻甲氧基苯甲酸）或峰高（无内标）在313nm处通过紫外检测进行定量。该方法可在25分钟内完成测定。龙胆酸的灵敏度为0.2mg/L，水杨尿酸和水杨酸的灵敏度均为0.5mg/L（进样体积20微升）；水杨酸及其代谢物的响应与浓度呈线性关系，至少可达2.000g/L。从尿液中回收水杨酸及其代谢物的分析回收率是完全的。水杨酸在7.5mg/L时的批内精密度（变异系数）为5.52%，水杨尿酸在9.33mg/L时为5.01%，龙胆酸在7.96mg/L时为3.07%。批间精密度方面，水杨酸在7.51mg/L时为7.32%，水杨尿酸在8.58mg/L时为5.52%，龙胆酸在8.32mg/L时为3.97%。我们发现，除阿司匹林外，接受各种药物治疗的患者尿液中没有明显干扰。
