DEVA Holding A.S., R&D Center, Karaağaç Mh. Fatih Blv. No: 26, Kapaklı 59510, Tekirdağ, Türkiye; Marmara University Institute of Health Science, İstanbul, Türkiye.
DEVA Holding A.S., R&D Center, Karaağaç Mh. Fatih Blv. No: 26, Kapaklı 59510, Tekirdağ, Türkiye.
J Pharm Biomed Anal. 2024 Jan 5;237:115761. doi: 10.1016/j.jpba.2023.115761. Epub 2023 Oct 2.
Ricobendazole hydrochloride is an active ingredient of a veterinary antiparasitic drug. The aim of this study was to investigate the degradation of ricobendazole hydrochloride under stress and stability testing conditions, for which we developed and validated the first stability indicating, specific, precise, accurate, and robust assay and related substances UPLC methods. The Acquity UPLC BEH C column was used for the related substances and assay analyses of ricobendazole hydrochloride, and the analyses were performed at 25 °C sample and 30 °C column temperatures with a 2 µL injection volume. In both methods, a mixture of water and methanol (60:40, v/v) was used as the diluent, mobile phase A was a phosphate buffer (50 mM potassium dihydrogen phosphate solution, pH 3.2 ± 0.05, adjusted with 10% o-phosphoric acid), and mobile phase B was a mixture of mobile phase A and acetonitrile (50:50, v/v). For the analysis of related substances, a gradient elution system was used at a flow rate of 0.4 mL/min for 35 min with a detection wavelength of 220 nm, while for the assay analysis; a gradient elution system was used at a flow rate of 0.3 mL/min for 15 min with a detection wavelength of 290 nm. The calibration curves showed excellent linearity with high R-squared (R) values for each compound, ricobendazole (0.9998, 0.249 - 3.740 µg/mL), ricobendazole amine (0.9998, 0.255 - 3.819 µg/mL), albendazole (0.9998, 0.255 - 7.646 µg/mL), and albendazole sulfone (1.0000, 0.251 - 15.090 µg/mL) indicating a strong correlation between the concentrations of the compounds and their respective peak areas in the UPLC analysis. The method showed excellent accuracy with relative standard deviation values of less than 2.5%. The stress and photostability studies showed that ricobendazole hydrochloride was insensitive to daylight and UV radiation and showed significant degradation at elevated temperature (85 °C, 9 days) and under all hydrolysis and oxidation conditions. The major impurity was ricobendazole amine under thermal and hydrolysis conditions, while albendazole sulfone was the major oxidative impurity.
盐酸甲苯咪唑是一种兽医驱虫药的有效成分。本研究旨在考察盐酸甲苯咪唑在各种应激和稳定性试验条件下的降解情况，为此我们开发并验证了首个专属性强、精确、准确、灵敏且稳健的含量测定和有关物质的 UPLC 方法。使用 Acquity UPLC BEH C 柱对盐酸甲苯咪唑的有关物质和含量测定进行分析，在 25°C 进样和 30°C 柱温下，以 2µL 进样量进行分析。在这两种方法中，均使用水和甲醇（60:40，v/v）混合溶液作为稀释剂，流动相 A 为 50mM 磷酸二氢钾溶液（pH3.2±0.05，用 10% 磷酸调节），流动相 B 为流动相 A 和乙腈（50:50，v/v）的混合物。有关物质分析采用梯度洗脱系统，流速为 0.4mL/min，洗脱时间为 35min，检测波长为 220nm；而含量测定分析采用梯度洗脱系统，流速为 0.3mL/min，洗脱时间为 15min，检测波长为 290nm。校准曲线显示各化合物均具有极好的线性关系，相关系数（R）值均很高，甲苯咪唑（0.9998，249-3740μg/mL）、甲苯咪唑胺（0.9998，255-3819μg/mL）、阿苯达唑（0.9998，255-7646μg/mL）和阿苯达唑砜（1.0000，251-15090μg/mL）。这表明化合物浓度与其在 UPLC 分析中的相应峰面积之间存在很强的相关性。该方法的精密度非常好，相对标准偏差值均小于 2.5%。稳定性研究表明，盐酸甲苯咪唑对日光和紫外线不敏感，在高温（85°C，9 天）和所有水解及氧化条件下均有明显降解。在热和水解条件下，主要杂质为甲苯咪唑胺，而氧化条件下的主要杂质为阿苯达唑砜。
