College of Chemistry and Environmental Science, Hebei University, Key Laboratory of Analytical Science and Technology of Hebei Province, Baoding 071002, China.
Drug Test Anal. 2009 Feb;1(2):87-92. doi: 10.1002/dta.22.
The content of free proline (Pro) in body fluids is a biological parameter for patients with renal insufficiency and chronic uraemia. Fleroxacin (FLX) must be used cautiously because of adverse reactions. Its dosage must be adjusted according to the degree of renal insufficiency. In a clinical setting the simultaneous determination of Pro and FLX in body fluids is necessary for the rational utilization of FLX. A capillary electrophoresis (CE) method based on electrochemiluminescence (ECL) detection with Ru(bpy)(3) (2+) was developed for the simultaneous determination of Pro and FLX in human urine. Parameters related to separation and detection were investigated and optimized. The most favourable resolution and high sensitivity were obtained using a 15 mM phosphate buffer at pH 9.6 with the detection potential at 1.15 V. Under optimized conditions, the standard curves were linear in the range of 0.1-80 microg/mL for Pro and 0.1-100 microg/mL for FLX. Detection limits(3sigma) of 0.3 ng/mL for FLX and 0.02 microg/mL for Pro were obtained. Relative standard derivations (RSDs) of the ECL intensity and the migration time were 3.2% and 0.9% for 4 microg/mL Pro and 3.7% and 1.2% for 4 microg/mL FLX, respectively. The recoveries were in the range of 94.8-99.6% for Pro with RSDs of 2.8%-3.6% and 94.7-97.8% for FLX with RSDs of 2.9%-3.7%. The proposed method was successfully applied to determine the amounts of Pro and FLX in human urine.
体液中游离脯氨酸（Pro）的含量是肾功能不全和慢性尿毒症患者的生物学参数。由于不良反应，氟罗沙星（FLX）必须谨慎使用。其剂量必须根据肾功能不全的程度进行调整。在临床环境中，同时测定体液中的 Pro 和 FLX 对于 FLX 的合理利用是必要的。建立了一种基于电化学发光（ECL）检测的毛细管电泳（CE）法，用于同时测定人尿中的 Pro 和 FLX。研究并优化了与分离和检测相关的参数。在 pH 值为 9.6 的 15 mM 磷酸盐缓冲液中，检测电位为 1.15 V 时，获得了最佳的分辨率和高灵敏度。在优化条件下，Pro 的标准曲线在 0.1-80 µg/mL 范围内线性，FLX 的标准曲线在 0.1-100 µg/mL 范围内线性。FLX 的检测限（3sigma）为 0.3 ng/mL，Pro 的检测限（3sigma）为 0.02 µg/mL。对于 4 µg/mL Pro，ECL 强度和迁移时间的相对标准偏差（RSD）分别为 3.2%和 0.9%，对于 4 µg/mL FLX，RSD 分别为 3.7%和 1.2%。Pro 的回收率在 94.8-99.6%之间，RSD 为 2.8%-3.6%，FLX 的回收率在 94.7-97.8%之间，RSD 为 2.9%-3.7%。该方法成功应用于测定人尿中 Pro 和 FLX 的含量。
