Two high-performance liquid chromatography-diode array detection methods have been developed and validated for the simultaneous quantification of genistein (GNS) and all trans retinoic acid (ATRA) as a novel anticancer combination therapy in their co-formulated nanoparticles and in rat plasma. Separation was performed on C18 column (250 × 4.6 mm, 5 μm) using celecoxib as internal standard. A mobile phase containing acetonitrile and water adjusted to pH 3 using 1% trifluoroacetic acid was delivered in gradient elution modes with time programmed UV detection. For extraction of the drugs and the internal standard from rat plasma, liquid- liquid extraction was applied. The proposed methods were validated as per International Conference on Harmonisation (ICH) guidelines (in the range 0.1-10 μg/mL for analysis of GNS and ATRA in nanoparticles) or according to Food and Drug Administration (FDA) guidance on bioanalytical method validation (in the range 0.025-20 μg/mL for analysis of GNS and ATRA in rat plasma). Pharmacokinetic study in six rats was performed following intravenous (IV) administration of a single dose of 0.5 mg/Kg of GNS and ATRA. The drugs' concentrations were measured up to 24 hours, and different pharmacokinetic parameters were calculated. The obtained parameters were comparable with the reported values for IV administration of each drug alone in rats. This confirms the applicability of the proposed method in monitoring the levels of the two drugs in vivo following their coadministration and indicating that the two drugs could be coadministered as a promising novel combination therapy for the treatment of lung cancer without great alteration in their pharmacokinetic parameters compared with their individual IV administration.

中文翻译：

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灵敏廉价的HPLC测定纳米颗粒和大鼠血浆中新型抗癌药物的组合：药代动力学应用。

已经开发了两种高效液相色谱-二极管阵列检测方法，并验证了其同时配制的纳米颗粒和大鼠血浆中染料木黄酮（GNS）和所有反式维甲酸（ATRA）的同时定量作为一种新型的抗癌联合疗法。使用塞来昔布作为内标在C18色谱柱（250×4.6 mm，5μm）上进行分离。含有乙腈和水（已使用1％三氟乙酸调节至pH 3）的流动相以梯度洗脱模式提供，并带有程序编程的紫外线检测功能。为了从大鼠血浆中提取药物和内标，应用了液-液提取。所提出的方法已根据国际协调会议（ICH）指南进行了验证（范围为0。用于分析纳米颗粒中的GNS和ATRA的1-10μg/ mL）或根据食品和药物管理局（FDA）关于生物分析方法验证的指导（用于分析大鼠血浆中的GNS和ATRA的范围为0.025-20μg/ mL） 。在静脉（IV）给予0.5 mg / Kg的GNS和ATRA单剂量后，在六只大鼠中进行了药代动力学研究。在长达24小时的时间内测量药物的浓度，并计算出不同的药代动力学参数。所获得的参数与大鼠中每种药物单独静脉给药的报道值相当。