Ahmed Hytham M, Ebeid Wael B
Pharmaceutical Analysis Department, Faculty of Pharmacy, Damanhour University, Damanhour, Egypt.
SEDICO Pharmaceuticals, Merck & Co External Partner, 6th of October City, Cairo, Egypt.
Spectrochim Acta A Mol Biomol Spectrosc. 2015 May 15;143:12-9. doi: 10.1016/j.saa.2015.02.025. Epub 2015 Feb 14.
Complex samples analysis is a challenge in pharmaceutical and biopharmaceutical analysis. In this work, tobramycin (TOB) analysis in human urine samples and recombinant human erythropoietin (rhEPO) analysis in the presence of similar protein were selected as representative examples of such samples analysis. Assays of TOB in urine samples are difficult because of poor detectability. Therefore laser induced fluorescence detector (LIF) was combined with a separation technique, micellar electrokinetic chromatography (MEKC), to determine TOB through derivatization with fluorescein isothiocyanate (FITC). Borate was used as background electrolyte (BGE) with negative-charged mixed micelles as additive. The method was successively applied to urine samples. The LOD and LOQ for Tobramycin in urine were 90 and 200ng/ml respectively and recovery was >98% (n=5). All urine samples were analyzed by direct injection without sample pre-treatment. Another use of hyphenated analytical technique, capillary zone electrophoresis (CZE) connected to ultraviolet (UV) detector was also used for sensitive analysis of rhEPO at low levels (2000IU) in the presence of large amount of human serum albumin (HSA). Analysis of rhEPO was achieved by the use of the electrokinetic injection (EI) with discontinuous buffers. Phosphate buffer was used as BGE with metal ions as additive. The proposed method can be used for the estimation of large number of quality control rhEPO samples in a short period.
复杂样品分析是药物和生物制药分析中的一项挑战。在本研究中，选择人尿液样品中的妥布霉素（TOB）分析以及在存在相似蛋白质情况下的重组人促红细胞生成素（rhEPO）分析作为此类样品分析的代表性实例。尿液样品中TOB的检测由于可检测性差而颇具难度。因此，将激光诱导荧光检测器（LIF）与分离技术胶束电动色谱（MEKC）相结合，通过异硫氰酸荧光素（FITC）衍生化来测定TOB。以硼酸盐作为背景电解质（BGE），带负电荷的混合胶束作为添加剂。该方法相继应用于尿液样品。尿液中妥布霉素的检测限和定量限分别为90和200ng/ml，回收率>98%（n = 5）。所有尿液样品均不经样品预处理直接进样分析。联用分析技术的另一个应用，即连接紫外（UV）检测器的毛细管区带电泳（CZE），也用于在大量人血清白蛋白（HSA）存在的情况下对低水平（2000IU）的rhEPO进行灵敏分析。rhEPO的分析通过使用带有不连续缓冲液的电动进样（EI）来实现。以磷酸盐缓冲液作为BGE，金属离子作为添加剂。所提出的方法可用于在短时间内对大量rhEPO质量控制样品进行评估。
