Determination of binded and free formaldehyde in cross-linked sodium hyaluronate injection by high performance liquid chromatography with pre-column derivatization / 中国药学杂志

ABSTRACT

OBJECTIVE: To develop a high performance liquid chromatography method with pre-column derivatization for the determination of binded and free formaldehyde in cross linked sodium hyaluronate injection. METHODS: After the samples were hydrolyzed by heating with 1.0 mol · L-1 hydrochloric acid, binded formaldehyde was released. By contrast, the concentration of free formaldehyde was measured without the hydrolysis step. Formaldehyde was derived with 2, 4-dinitrophenylhydrazine (DNPH) and then determined by HPLC. The analysis was carried out on a C18 column(4.6 mm × 250 mm, 5 μm). The mobile phase was 65% acetonitrile, the flow rate was 1.0 mL · min-1, and the wavelength was set at 365 nm. RESULTS: This method had a linear range of 0.1-50 μg · mL-1 (y =76 023x +7 069, r=1.0000). The limit of detection(LOD) and the limit of quantification(LOQ) were 0.001 μg · mL-1 and 0.005 μg · mL-1, respectively. The average recoveries of the binded and free formaldehyde were 102.9% and 105.5%, and the relative standard deviations (RSDs, n=18) were 1.3% and 3.0%, respectively. CONCLUSION: This method has a wide linear range and good accuracy, which is suitable for the determination of both binded and free formaldehyde contents in cross linked sodium hyaluronate injection.