Institute of Health Sciences, Universidade Feevale, Novo Hamburgo, Brazil.
Clin Biochem. 2013 Sep;46(13-14):1276-80. doi: 10.1016/j.clinbiochem.2013.07.004. Epub 2013 Jul 15.
The purpose of this study was to develop and validate a HPLC-DAD method for determination of creatinine and hippuric acid in dried urine spots (DUS), evaluating its clinical applicability.
Sample preparation was based on a simple one step extraction with water. Analysis was performed in a reversed phase column Hypersil Gold C18 (150 × 4.6 mm, 3 μm) with isocratic elution. Mobile phase was a mixture of triethylammonium phosphate buffer 5 mM, pH 3.3 and acetonitrile (90:10, v/v).
Total analytical run time was 6 min. Precision assays presented CV% lower than 11.8%. Accuracy was 96.0-103%. The lower limit of quantitation was 50 mg L(-1). Mean extraction yields were 99% for CR and 79.3% for HA. Analytes were stable in DUS up to 11 days at 40 °C. The method was tested in 49 paired DUS and urine samples, with acceptable agreement. HA-CR concentration ratios in DUS were in the range of 0.06 to 2.74, being in average 100.7% from those values found in urine.
DUS samples are a useful alternative for biological monitoring of toluene occupational exposure, especially in Developing Countries where sample logistics could be an important concern.
本研究旨在开发和验证一种 HPLC-DAD 方法，用于测定干尿斑（DUS）中的肌酐和马尿酸，并评估其临床适用性。
样品制备基于简单的一步水提取。分析在 Hypersil Gold C18 反相柱（150×4.6mm，3μm）上进行等度洗脱。流动相为 5mM 三乙胺磷酸盐缓冲液，pH3.3 和乙腈（90:10，v/v）的混合物。
总分析运行时间为 6 分钟。精密度试验的 CV%低于 11.8%。准确度为 96.0-103%。定量下限为 50mgL(-1)。CR 的平均提取回收率为 99%，HA 的平均提取回收率为 79.3%。在 40°C 下，DUS 中分析物稳定达 11 天。该方法在 49 对 DUS 和尿液样品中进行了测试，具有可接受的一致性。DUS 中 HA-CR 浓度比在 0.06 至 2.74 之间，平均为尿液中值的 100.7%。
DUS 样本是甲苯职业暴露生物监测的一种有用替代方法，特别是在样品物流可能是一个重要关注点的发展中国家。
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