Manouilov K K, McGuire T R, Gwilt P R
UNMC/Eppley Cancer Center Core Pharmacokinetics Laboratory, Department of Pharmaceutical Sciences, College of Pharmacy, The University of Nebraska Medical Center, Omaha 69198-6025, USA.
J Chromatogr B Biomed Sci Appl. 1998 Apr 24;708(1-2):321-4. doi: 10.1016/s0378-4347(97)00634-8.
A high-performance liquid chromatography assay for hydroxyurea in human serum was developed based on a commercial colorimetric assay kit for urea (Sigma Diagnostics). Serum (0.5 ml), spiked with methylurea as an internal standard, was treated with 70% perchloric acid. Supernatant (0.2 ml) was combined with 0.7 ml of BUN acid reagent and 0.6 ml of BUN color reagent. The resulting colored reactant (100 microl) was analyzed on a 300 x 3.9 mm Bondclone 10 C18 column coupled with a UV-Vis detector, at 449 nm. The mobile phase was 13% acetonitrile in water. Retention times of colored derivatives of hydroxyurea and methylurea were 6.5 and 12.2 min, respectively. The log-log calibration curve was linear from 0.0065 to 1.31 mM. Average accuracy was 99.9+/-4.0% and the intra- and inter-day error of assay did not exceed 11%.
基于一种商业化的尿素比色检测试剂盒（Sigma诊断公司），开发了一种用于检测人血清中羟基脲的高效液相色谱分析法。向0.5 ml血清中加入甲基脲作为内标，然后用70%的高氯酸进行处理。取0.2 ml上清液与0.7 ml尿素氮酸性试剂和0.6 ml尿素氮显色试剂混合。将得到的有色反应物（100微升）在一根300×3.9 mm的Bondclone 10 C18柱上进行分析，该柱与紫外可见检测器联用，检测波长为449 nm。流动相为13%乙腈的水溶液。羟基脲和甲基脲有色衍生物的保留时间分别为6.5分钟和12.2分钟。对数-对数校准曲线在0.0065至1.31 mM范围内呈线性。平均准确度为99.9±4.0%，该分析方法的日内和日间误差均不超过11%。
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