背景:Flibanserin(FLB)最初是作为抗抑郁药合成的;然而,由于它对性活动的增强作用,该药物于2015年被批准用于女性性欲减退的治疗.
目的:本研究的目的是开发一种新的,经过充分验证的HPLC方法,用于分析药物制剂中除降解产物外的FLB,并通过使用HPLC-DAD-ESI-IT-TOF-MSn鉴定可能的形成机理。
方法:在SupelcoAscentis®Express系列苯基己基柱(100×4.6mm,ID2.7µm)。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的速率泵送。色谱,使用LCMS-IT-TOF仪器(Shimadzu,日本)。
结果: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-酮被提议作为新型降解产物,质量为407.1695,配方为C20H21F3N4O2,误差幅度约为0.001ppm。所开发的方法在2.5-50.0µg/mL范围内适用,准确度为98%。LOD和LOQ约为500ng/mL和1.50µg/mL,分别。通过实验室间比较测试实验室之间的可转移性和差异,并通过单向方差分析进行评估。
结论:一种新型的FLB降解产物,首次观察并鉴定了在氧化强制降解条件下产生的降解产物的形成动力学,此外还对FLB的分解进行了研究。此外,进行了实验室间比较,并使用两种仪器配置测试了拟议方法在伪Addyi®样品上的应用。
结论:根据ICH(Q2)R1开发并完全验证了一种新的稳定性指示测定方法,用于分析药物制剂中的FLB。在氧化强制降解条件下鉴定了新的降解产物,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理和形成动力学。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,将目前的方法应用于两种仪器中的Addy®的伪制剂,并应用ANOVA进行评估.
BACKGROUND: 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.
OBJECTIVE: 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.
METHODS: 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).
RESULTS: 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.
CONCLUSIONS: 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.
CONCLUSIONS: 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.