Robust In Vitro HPLC-DAD Method for Accurate Quantification of N-Carbamylglutamate in Drug Formulations

A comprehensive in vitro testing method has been developed and validated for accurately quantifying N-Carbamylglutamate (Carglumic acid) using a high-performance liquid chromatography instrument with a diode array detector. A method specifically developed for pharmaceutical quality control utilizes a reverse-phase C18 column for precise and reliable measurement of Carglumic acid. The chromatographic parameters were refined for peak resolution, employing a phosphate buffer and acetonitrile as the mobile phase, with detection setup at 205 nm. The method was validated under ICH Q2(R2) requirements. The approach exhibited a significant level of specificity, with no interference from excipients or other possible impurities. Linearity was achieved across the entire concentration range, yielding an R² value of 0.999. Precision (RSD ≤ 2) was demonstrated by intra- and inter-day variability being within acceptable limits. Recovery tests validated accuracy (98 – 102%). Determining the limit of detection (LOD, 0.1 ppm) and the limit of quantitation (LOQ, 0.5 ppm) was conducted. This in vitro test method was valid and effective for the routine quantification of Carglumic acidin pharmaceutical formulations, serving as a crucial tool for ensuring product quality and regulatory compliance. The methods' elevated sensitivity and endurance make it appropriate for bioanalytical applications, such as dissolution testing and stability analysis.