Clinical Pharmacy, College of Pharmacy, King Saud University, P.O.Box 2457, Riyadh 11451, Saudi Arabia. b
Talanta. 2010 Nov 15;83(1):110-6. doi: 10.1016/j.talanta.2010.08.047. Epub 2010 Sep 22.
A highly selective, sensitive and rapid high performance liquid chromatographic method has been developed and validated to quantify gemifloxacin in human plasma. The gemifloxacin and internal standard (ciprofloxacin) were extracted by ultrafiltration technique followed by injection into chromatographic system. Chromatographic separation was achieved on a reversed phase C(18) column with a mobile phase of acetonitrile:0.1% trifluoroacetic acid (20:80, v/v) using isocratic elution (at flow rate 1 mL min(-1)). The analytes were detected at 269 and 393 nm for excitation and emission, respectively. The assay exhibited a linear range of 25-5000 ng mL(-1) for gemifloxacin in human plasma. The lower limit of detection was 10 ng mL(-1). The method was statistically validated for linearity, accuracy, precision and selectivity following FDA guidelines. The intra- and inter-assay coefficients of variation did not exceed 7.6% deviation of the nominal concentration. The recovery of gemifloxacin from plasma was greater than 97.0%. Stability of gemifloxacin in plasma was excellent with no evidence of degradation during sample processing (auto-sampler) and at least 3 months storage in a freezer at -70 °C. This validation method is applied for clinical study of the gemifloxacin in human volunteers.
一种高选择性、高灵敏度和快速的高效液相色谱法已经被开发和验证，用于定量人血浆中的加替沙星。加替沙星和内标（环丙沙星）通过超滤技术提取，然后注入色谱系统。色谱分离在反相 C(18)柱上进行，流动相为乙腈：0.1%三氟乙酸（20:80，v/v），采用等度洗脱（流速为 1 mL min(-1)）。分析物在 269nm 和 393nm 处分别进行激发和发射检测。该测定法在人血浆中加替沙星的线性范围为 25-5000ng mL(-1)。检测限为 10ng mL(-1)。该方法按照 FDA 指南进行了线性、准确性、精密度和选择性的统计验证。日内和日间变异系数不超过名义浓度的 7.6%偏差。加替沙星从血浆中的回收率大于 97.0%。加替沙星在血浆中的稳定性非常好，在样品处理（自动进样器）过程中和至少在-70°C 的冰箱中储存 3 个月内没有降解的证据。该验证方法应用于人体志愿者的加替沙星临床研究。
