Abdel-Aziz Heba, Tolba M M, El-Enany N, Aly F A, Fathy M E
Department of Pharmaceutical Analytical Chemistry, Faculty of Pharmacy, Mansoura University, 35516 Mansoura, Egypt.
R Soc Open Sci. 2021 Aug 4;8(8):210329. doi: 10.1098/rsos.210329. eCollection 2021 Aug.
Using two green and sensitive spectrofluorimetric methods, we quantified two cephalosporins, cefepime (CFM) and cefazolin (CFZ), in raw and pharmaceutical formulations. The first method is based on the reaction between CFM and fluorescamine (borate buffer, pH 8), which yields a highly fluorescent product. After excitation at 384 nm, the fluorescent product emits light at 484 nm. At concentration ranges from 12.0 to 120.0 ng ml, the relative fluorescence intensity/concentration curve was linear with a limit of quantification (LOQ) of 2.46 ng ml. The second method relied on measuring the CFZ quenching action on acriflavine fluorescence through formation of an ion-associate complex using Britton-Robinson buffer at pH 8. We measured acriflavine fluorescence at 505 nm after excitation at 265 nm. The decrease in acriflavine fluorescence intensity was CFZ concentration-dependent. Using this method, we quantified CFZ in concentrations ranging from 1 to 10 µg ml with a LOQ of 0.48 µg ml. We studied and optimized the factors influencing reaction product formation. Moreover, we adapted our methods to the investigation of the mentioned drugs in raw and pharmaceutical formulations with greatly satisfying results. We statistically validated our methods according to International Council on Harmonisation Guidelines. The obtained results were consistent with those obtained with the official high-performance liquid chromatography methods.
我们使用两种绿色且灵敏的荧光光谱法,对原料药和药物制剂中的两种头孢菌素,即头孢吡肟(CFM)和头孢唑林(CFZ)进行了定量分析。第一种方法基于CFM与荧光胺(硼酸盐缓冲液,pH 8)之间的反应,该反应产生一种高荧光产物。在384 nm激发后,该荧光产物在484 nm处发光。在12.0至120.0 ng/ml的浓度范围内,相对荧光强度/浓度曲线呈线性,定量限(LOQ)为2.46 ng/ml。第二种方法依靠在pH 8的 Britton-Robinson缓冲液中通过形成离子缔合物络合物来测量CFZ对吖啶黄荧光的猝灭作用。在265 nm激发后,我们在505 nm处测量吖啶黄荧光。吖啶黄荧光强度的降低与CFZ浓度相关。使用该方法,我们对浓度范围为1至10 µg/ml的CFZ进行了定量,定量限为0.48 µg/ml。我们研究并优化了影响反应产物形成的因素。此外,我们将我们的方法应用于原料药和药物制剂中上述药物的研究,结果非常令人满意。我们根据国际协调理事会指南对我们的方法进行了统计学验证。所得结果与官方高效液相色谱法所得结果一致。