Global Pharmaceutical Technical Support. , Boehringer Ingelheim Animal Health USA Inc. (BIAH) 631 US Route 1 South, North Brunswick, NJ United States.
J AOAC Int. 2024 Jul 4;107(4):600-607. doi: 10.1093/jaoacint/qsae027.
Fipronil is a commonly used pesticide in the agricultural and animal health industries for the protection of crops and control of fleas, ticks, and chewing lice. It is difficult to obtain reproducible retention time and relative retention time (RRT) for a common hydrolytic degradation product of fipronil with the current European Pharmacopeia (EP) monograph for assay and estimation of related substances of fipronil. This situation causes misidentification, mislabeling, and/or false out-of-specification results for this hydrolytic degradation product of fipronil in bulk commercial batches during batch release and/or in the stability samples during the shelf life of a released batch.
This study aimed to develop a reversed-phase ultra performance liquid chromatography (UPLC) method for assay and identification of fipronil including identification and estimation of its related substances in bulk drug substance batches of fipronil and provide consistent retention time of the hydrolytic degradation product.
Fipronil and its related substances were separated by gradient elution on a Halo C18 column (50 mm × 2.1 mm id, 2.0 µm particle size) maintained at 40°C with 0.1% H3PO4 in H2O as mobile phase-A and acetonitrile-methanol (50 + 50, v/v) as mobile phase-B. Fipronil and its related substances were detected and quantified at 280 nm with a quantitation limit of 0.05% of the target (analytical) concentration.
The UPLC method was able to separate all analytes of interest by gradient elution with a total run time of 7 min (approximately 40% faster than EP).
In this paper, we report the development and validation of a fast, stability-indicating reversed-phase UPLC method for assay and estimation of related substances of fipronil in stability samples and bulk batches of fipronil.
The new UPLC method is approximately 40% faster than the current Ph. Eur. monograph for fipronil assay and the new method provides reproducible retention of a common hydrolytic degradation product of fipronil.
氟虫腈是农业和动物保健行业中常用的杀虫剂，用于保护作物和控制跳蚤、蜱和虱子。目前欧洲药典（EP）的氟虫腈测定和有关物质评估专论中，对于氟虫腈的一种常见水解降解产物，难以获得重现的保留时间和相对保留时间（RRT）。这种情况会导致在批量放行时的商业批次大量药物中，以及在放行批次的保质期内的稳定性样品中，对这种氟虫腈水解降解产物进行错误识别、错误贴标和/或误判超出规格。
本研究旨在开发一种反相超高效液相色谱（UPLC）方法，用于测定氟虫腈及其有关物质，包括鉴定和评估氟虫腈原料药批次中的相关物质，并提供水解降解产物的一致保留时间。
在 Halo C18 柱（50mm×2.1mm id，2.0μm 粒径）上，以 0.1%H3PO4在 H2O 中作为流动相-A和乙腈-甲醇（50+50，v/v）作为流动相-B进行梯度洗脱，分离氟虫腈及其有关物质。采用 280nm 处的检测和定量，定量限为目标（分析）浓度的 0.05%。
该 UPLC 方法能够通过梯度洗脱分离所有感兴趣的分析物，总运行时间为 7 分钟（比 EP 快约 40%）。
本文报道了一种快速、稳定指示的反相 UPLC 方法的开发和验证，用于测定和评估氟虫腈在稳定性样品和氟虫腈原料药批量中的有关物质。
新的 UPLC 方法比目前的欧洲药典专论中氟虫腈测定法快约 40%，并且该新方法提供了氟虫腈常见水解降解产物的重现保留。
