Elzanfaly Eman Saad, Amer Enas Abdel Hakim, Galal Sara Abdel Basset, Zaazaa Hala Elsayed
Analytical Chemistry Department, Faculty of Pharmacy, Cairo University, Kasr El- Aini Street, Cairo, Egypt.
National Organization for Drug Control and Research (NODCAR), 51 Wezerat El- Zeraa Street, Agouza, Giza, Egypt.
Luminescence. 2019 Feb;34(1):64-69. doi: 10.1002/bio.3577. Epub 2018 Dec 11.
A spectrofluorimetric method for the determination of eptifibatide is presented based on its native fluorescence. The type of solvent and the wavelength of maximum excitation and emission were carefully selected to optimize the experimental conditions. Under the specified experimental conditions, the linearities obtained between the emission intensity and the corresponding concentrations of eptifibatide were in the range 0.1-2.5 μg/ml for the calibration curve constructed for direct determination of eptifibatide in dosage form and 0.05-2.2 μg/ml for the calibration curve constructed in spiked human plasma with a good correlation coefficient (r > 0.99). The lower limit of quantification for the calibration curve constructed in human plasma was 0.05 μg/ml. Recovery results for eptifibatide in spiked plasma samples and in dosage form, represented as mean ± % RSD, were 95.17 ± 1.94 and 100.29 ± 1.33 respectively. The suggested procedures were validated according to the International Conference on Harmonization (ICH) guidelines for the direct determination of eptifibatide in its pure form and dosage form and United States Food and Drug Administration (US FDA) Guidance for Industry, Bioanalytical Method Validation for the assay of eptifibatide in human plasma.
提出了一种基于依替巴肽自身荧光来测定其含量的荧光分光光度法。仔细选择了溶剂类型以及最大激发波长和发射波长，以优化实验条件。在规定的实验条件下，对于直接测定剂型中依替巴肽所构建的校准曲线，发射强度与依替巴肽相应浓度之间的线性范围为0.1 - 2.5μg/ml；对于在加标人血浆中构建的校准曲线，线性范围为0.05 - 2.2μg/ml，相关系数良好（r > 0.99）。在人血浆中构建的校准曲线的定量下限为0.05μg/ml。加标血浆样品和剂型中依替巴肽的回收率结果，以平均值±%相对标准偏差表示，分别为95.17 ± 1.94和100.29 ± 1.33。根据国际协调会议（ICH）关于直接测定纯品和剂型中依替巴肽的指南以及美国食品药品监督管理局（US FDA）关于人血浆中依替巴肽测定的生物分析方法验证的行业指南，对所建议的方法进行了验证。
