Determination of Four Related Substances in Tegoprazan by HPLC

ABSTRACT In this study, a main component self‐comparison method with relative correction factors by high‐performance liquid chromatography (HPLC) was developed and validated for detecting or quantifying the four impurities in tegoprazan. The chromatographic separation was performed on a CAPCELL CORE C 18 (4.6 mm × 75 mm, 2.7 µm) column in the gradient mode by using 0.01 mol/L ammonium dihydrogen phosphate solution (the pH value adjusted to 6.5 with ammonia solution) and acetonitrile as the eluent. The flow rate was 0.7 mL/min, the detection wavelength was set at 218 nm, and the column temperature was maintained at 45°C. Under the optimized conditions, related substances and tegoprazan were completely separated. The linear ranges for the four related substances were impurity A 0.1071–10.71 µg/mL ( r = 1.000), impurity B 0.1095–10.95 µg/mL ( r = 1.000), impurity C 0.0961–9.61 µg/mL ( r = 0.9999), and impurity D 0.1152–11.52 µg/mL ( r = 0.9999). The average recoveries were 96.47%–97.17%, 93.51%–95.60%, 93.90%–95.95%, and 93.39%–95.40%, respectively, with RSDs all less than 2.0%. With reference to tegoprazan (retention time = 13.8 min), the relative retention times for impurities A, B, C, and D were around 0.22, 0.73, 0.86, and 1.86, respectively, and the relative correction factors were 0.78, 1.17, 3.16, and 1.19, respectively. The proposed method was applied to assess different commercially available tegoprazan samples, and any of the contents of impurities A, B, C, and D could not be more than 0.78 mg/g, which was consistent with the standard curve method. This method was proven to be accurate, reliable, highly sensitive, and suitable for quality control of four related substances in tegoprazan.