Anadolu University, Faculty of Pharmacy, Department of Analytical Chemistry, 26470 Eskisehir, Türkiye.
Anadolu University, Faculty of Pharmacy, Central Analysis Laboratory, 26470 Eskişehir, Türkiye.
J AOAC Int. 2023 Sep 1;106(5):1145-1153. doi: 10.1093/jaoacint/qsad074.
Flibanserin (FLB) was first synthesized as an antidepressant drug; however, due to its enhancing effects on sexual activity, it was approved for treatment of hypoactive sexual desire disorder in women in 2015.
The aim of this study was to develop a new and fully validated HPLC method for analysis of FLB in pharmaceutical formulations besides its degradation products, and identification of possible formation mechanisms by using HPLC-DAD-ESI-IT-TOF-MSn.
The HPLC separation was achieved in a Supelco Ascentis® Express series phenyl hexyl column (100 × 4.6 mm, ID 2.7 µm). The mobile phase was acetonitrile-ammonium acetate solution (50:50, v/v, 10 mM, pH 5.4) mixture, which was pumped at the rate of 0.5 mL/min. Chromatography, detection, and structural identification was performed by using a LCMS-IT-TOF instrument (Shimadzu, Japan).
1-(2-(4-(3-hydroxy-5-(trifluoromethyl)phenyl)piperazine-1-yl)ethyl)-1,3-dihydro-2H-benzo[d]imidazol-2-one is proposed as a novel degradation product, with a mass of 407.1695 and a formula of C20H21F3N4O2 with a margin of error about 0.001 ppm. The developed method is applicable with 98% accuracy within the 2.5-50.0 µg/mL range. The LOD and LOQ were about 500 ng/mL and 1.50 µg/mL, respectively. The transferability and variation between laboratories were tested by inter-laboratory comparison and evaluated with one-way analysis of variance.
A novel FLB degradation product, which was produced under oxidative forced degradation conditions was observed and identified for the first time; in addition, the formation kinetics of the degradation product besides decomposition of FLB was studied. Furthermore, an inter-laboratory comparison was carried out, and application of the proposed method on a pseudo Addyi® (Sprout Pharmaceuticals, Inc.) sample was tested using both instrument configurations.
A novel stability-indicating assay method was developed and fully validated according to the International Council on Harmonization (Q2) R1 for the analysis of FLB in the pharmaceutical preparations. A new degradation product was identified in the oxidative forced degradation condition and characterized using HPLC-DAD-ESI-IT-TOF-MS3. Moreover, the possible mechanism and the formation kinetic of the degradation product were revealed. In addition, the developed method was transferred to another LC-PDA instrument for inter-laboratory comparison. Finally, the current method was applied to a pseudo formulation of Addy in both instruments, and ANOVA was applied for evaluation.
氟班色林（FLB）最初被合成作为一种抗抑郁药物; 然而，由于其增强性行为的作用，它于 2015 年被批准用于治疗女性性欲低下障碍。
本研究的目的是开发一种新的和完全验证的 HPLC 方法，用于分析药物制剂中的 FLB 及其降解产物，并通过使用 HPLC-DAD-ESI-IT-TOF-MSn 来鉴定可能的形成机制。
HPLC 分离在 Supelco Ascentis®Express 系列苯基己基柱（100×4.6mm，ID 2.7µm）上进行。流动相为乙腈-乙酸铵溶液（50：50，v/v，10mM，pH5.4）混合物，以 0.5mL/min 的速度泵入。通过 LCMS-IT-TOF 仪器（日本岛津公司）进行色谱、检测和结构鉴定。
提出了一种新的降解产物 1-（2-（4-（3-羟基-5-（三氟甲基）苯基）哌嗪-1-基）乙基）-1,3-二氢-2H-苯并[d]咪唑-2-酮，其质量为 407.1695，分子式为 C20H21F3N4O2，误差幅度约为 0.001ppm。开发的方法在 2.5-50.0µg/mL 范围内具有 98%的准确度。LOD 和 LOQ 分别约为 500ng/mL 和 1.50µg/mL。通过实验室间比较和单因素方差分析对转移性能和实验室间变异进行了测试。
首次观察到并鉴定了在氧化强制降解条件下产生的新型 FLB 降解产物; 此外，还研究了降解产物的形成动力学以及 FLB 的分解。此外，进行了实验室间比较，并使用两种仪器配置对伪 Addyi®（Sprout Pharmaceuticals，Inc.）样品进行了所提议方法的应用测试。
根据国际协调会议（Q2）R1 开发并全面验证了一种新型的稳定性指示测定方法，用于药物制剂中 FLB 的分析。在氧化强制降解条件下鉴定出一种新的降解产物，并使用 HPLC-DAD-ESI-IT-TOF-MS3 进行了表征。此外，揭示了降解产物的可能机制和形成动力学。此外，将开发的方法转移到另一台 LC-PDA 仪器进行实验室间比较。最后，在两种仪器上应用当前方法对 Addyi 的伪制剂进行了应用，并应用 ANOVA 进行了评估。
