Quantitative analysis of ripasudil hydrochloride hydrate and its impurities by reversed-phase high-performance liquid chromatography after precolumn derivatization: Identification of four impurities
| Autor: | Lili Sun, Liang Zou, Qiaogen Zou, Wenkai Hui, Pingkai Ouyang, Hui Zhang |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
Chromatography
Hydrochloride 010401 analytical chemistry Filtration and Separation 01 natural sciences High-performance liquid chromatography 0104 chemical sciences Analytical Chemistry 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine chemistry Reagent 030221 ophthalmology & optometry Methanol Ripasudil Hydrate Triethylamine Quantitative analysis (chemistry) |
| Zdroj: | Journal of Separation Science. 39:3302-3310 |
| ISSN: | 1615-9306 |
| Popis: | We report the development and validation of a stability-indicating reversed-phase high-performance liquid chromatography method with precolumn derivatization for the separation and identification of the impurities of ripasudil hydrochloride hydrate, a novel protein kinase inhibitor. 2,3,4,6-Tetra-O-acetyl-β-d-glucopyranosyl isothiocyanate was chosen as the derivatizing reagent and triethylamine was added as catalyst. 200 μL sample solution (1 mg/mL), 600 μL derivatizing reagent (1 mg/mL), and 200 μL triethylamine solution (1%, v/v) were mixed and reacted at 40°C for 30 min. The separation was achieved on an Inertsil C18 ODS-3 (250 mm × 4.6 mm, 5 μm) column using mobile phases including 10 mmol monopotassium phosphate buffer (pH 3.0) and methanol in gradient mode. The column temperature was adjusted at 25°C and the flow rate at 1 mL/min. The detection was carried out at 220 nm. Different precolumn derivatization conditions as well as the high-performance liquid chromatography conditions were optimized. Ripasudil hydrochloride hydrate and its four impurities were detected and quantitated, among which two new compounds were characterized. The proposed method was validated and proven to be selective, accurate, and precise and suitable for the quantitative analysis of ripasudil hydrochloride hydrate. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|