Abstract
This paper describes the development and validation a of reversed‐phase high performance liquid chromatography (RP‐HPLC) method for the assay of hydrolysed gelatine (Gelita‐Sol P, trade name). Key chromatographic parameters were investigated, including short and long alkyl chains of stationary phases (C4 and C18), column temperatures (30–60°C), and additives of ion‐pairing reagents (trifluoroacetic acid and heptafluorobutyric acid) in the mobile phase. Analytical validation parameters such as specificity and selectivity, linearity, accuracy, precision, limit of detection, limit of quantitation, robustness, and system suitability, were evaluated. The calibration curve for hydrolysed gelatine was linear (r 2=0.997) from 20–200% range of the analytical concentration of 50 mg/mL. The precision of this method calculated as the relative standard deviation (R.S.D.) was 1.22% (n=6). The R.S.D. for the intermediate precision study was 1.77, and recovery of the hydrolysed gelatine ranged between 97.08 and 97.76%. The limits of detection and quantitation were determined to be 5.0 and 10.0 mg/mL, respectively.
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