Karnes H T, Riley C M, Curry S H, Schulman S G
J Chromatogr. 1985 Mar 22;338(2):377-88. doi: 10.1016/0378-4347(85)80108-0.
A high-performance liquid chromatographic method is described for the analysis of bentiromide metabolites in urine. The procedure involves no more than direct injection of the diluted urine sample, obviating the need for an extraction step or an internal standard. A mu Bondapak C18 column is used with a mobile phase of 0.01 M tetrabutylammonium chloride (pH 7.4)--methanol (9:1). A flow-rate of 1.4 ml/min, detection at 254 nm and column temperature of 40 degrees C are employed. These conditions were achieved by investigating the effects of mobile phase pH, and concentrations and types of organic modifiers, buffers and ion-pairing agents on the resolution of the metabolites. The analysis time is 18 min per sample and the coefficient of variation on replicate assays is less than 10% for most concentrations studied. Analytical recoveries were between 95 and 100% throughout the appropriate concentration ranges and no interferences were obtained with the exception of p-acetamidobenzoyl glucuronide which could be eliminated by treatment of the samples with beta-glucuronidase. Concentration profiles of the metabolites were studied in normal subjects, and the method was found to be potentially useful for clinical situations in which the existing bentiromide test leads to ambiguous results because of small bowel and hepatic dysfunctions.
描述了一种用于分析尿液中苯替洛米代谢物的高效液相色谱法。该方法只需直接进样稀释后的尿液样品，无需萃取步骤或内标。使用μ Bondapak C18柱，流动相为0.01 M 四丁基氯化铵（pH 7.4）-甲醇（9:1）。流速为1.4 ml/min，检测波长为254 nm，柱温为40℃。通过研究流动相pH、有机改性剂、缓冲液和离子对试剂的浓度及类型对代谢物分离度的影响来确定这些条件。每个样品的分析时间为18分钟，在所研究的大多数浓度下，重复测定的变异系数小于10%。在适当的浓度范围内，分析回收率在95%至100%之间，除对乙酰氨基苯甲酰葡萄糖醛酸苷外无干扰，该物质可通过用β -葡萄糖醛酸苷酶处理样品来消除。在正常受试者中研究了代谢物的浓度分布，发现该方法对于因小肠和肝功能障碍导致现有苯替洛米试验结果不明确的临床情况可能有用。
