Development of a Method for Quantitative Determination of Carbohydrate Stabilizers in Biologics Based on HPLC with Refractometric Detection

A method for quantitative determination of carbohydrate stabilizers (sorbitol, mannitol, glucose, sucrose, lactose, maltose, and trehalose) in biologics was developed. The analysis used HPLC with refractometric detection and a mobile phase of 75% MeCN in H₂O. The developed method was specific and linear in the range 0.5 to 2 ng/mL. The coefficients of determination of the calibration curves were 0.99996 for sorbitol, 0.99998 for mannitol, 0.99971 for glucose, 0.99996 for sucrose, 0.99994 for lactose, 9.99966 for maltose monohydrate, and 0.99996 for trehalose dihydrate. The limits of quantitation (mg/mL) were 0.01 for sorbitol, 0.03 for mannitol, 0.05 for glucose, 0.025 for sucrose, 0.050 for lactose, 0.05 for maltose monohydrate, and 0.25 for trehalose dihydrate. The average accuracy covered the range (100 ± 5)%.