Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, Riyadh, Kingdom of Saudi Arabia.
Department of Analytical Chemistry Faculty of Pharmacy, Cairo University, Cairo, Egypt.
PLoS One. 2020 Dec 3;15(12):e0239918. doi: 10.1371/journal.pone.0239918. eCollection 2020.
This study describes, for the first time, the experimental and computational investigations for evaluation of kolliphor RH 40 as a fluorescence enhancer surfactant in development of a spectrofluorimetric method for determination of lapatinib (LAP), a tyrosine kinase-inhibitor drug approved for targeted therapy of breast cancer. The investigations involved the ability of kolliphor RH 40 to form micelles with LAP and its enhancing effect on the weak native fluorescence of LAP at 420 nm after its excitation at 292 nm. Different variables were experimentally investigated: types of organized media, diluting solvent, buffer type and its pH value. The optimum values of the most influencing variables on the interaction of kolliphor RH 40 with LAP were refined by the computational response surface methodology (RSM). Under the optimized conditions, it was found that kolliphor RH 40 forms micelles with LAP, and its fluorescence enhancing ability was higher than other surfactants tested by ~ 10-folds. This micellar-enhanced effect of kolliphor RH 40 was employed in the development of a new sensitive spectrofluorimetric method for the accurate determination of LAP. The method was validated according to the guidelines of the International Conference on Harmonization (ICH) for validation of analytical procedures. The relative fluorescence intensity (RFI) was in excellent linear relationship (correlation coefficient was 0.998) with the LAP concentrations in the range of 50-1000 ng/mL. The method limit of detection (LOD) was 27.31 ng/mL and its accuracy was ≥ 99.82%. The method was successfully applied to the determination of LAP in its pharmaceutical tablets, tablets dissolution testing and content uniformity. The method application was extended to the determination of LAP in urine samples with an accuracy of 99.82 ± 3.45%. The method is considered as an eco-friendly green approach and more efficient alternative method to the existing analytical methodologies for determination of LAP.
本研究首次描述了实验和计算研究，以评估 kolliphor RH 40 作为荧光增强表面活性剂在开发用于测定拉帕替尼（LAP）的分光荧光法中的应用，LAP 是一种批准用于乳腺癌靶向治疗的酪氨酸激酶抑制剂药物。研究涉及 kolliphor RH 40 与 LAP 形成胶束的能力及其在 292nm 激发后在 420nm 处增强 LAP 弱天然荧光的增强效果。实验研究了不同的变量：组织介质的类型、稀释溶剂、缓冲液类型及其 pH 值。通过计算响应面法（RSM）对影响 kolliphor RH 40 与 LAP 相互作用的最主要变量的最佳值进行了优化。在最佳条件下，发现 kolliphor RH 40 与 LAP 形成胶束，其荧光增强能力比其他测试的表面活性剂高约 10 倍。这种 kolliphor RH 40 的胶束增强效应被用于开发一种新的灵敏分光荧光法，用于准确测定 LAP。该方法根据国际协调会议（ICH）分析程序验证指南进行了验证。相对荧光强度（RFI）与 50-1000ng/mL 范围内的 LAP 浓度呈极好的线性关系（相关系数为 0.998）。该方法的检测限（LOD）为 27.31ng/mL，其准确度≥99.82%。该方法成功应用于其药物片剂中 LAP 的测定、片剂溶出度试验和含量均匀度。该方法的应用扩展到尿液样品中 LAP 的测定，准确度为 99.82±3.45%。该方法被认为是一种环保的绿色方法，是现有分析方法的更有效替代方法，用于测定 LAP。
