Objective: To establish a method for the determination of trehalose in monoclonal antibodies by high performance liquid chromatography-charged aerosol detector (HPLC-CAD). Methods: The chromatographic column was HyperREZ XP Carbohydrate Ca2+ (300 mm×7.7 mm, 8 μm), the mobile phase was ultra pure water, the flow rate was 0.8 mL·min-1, the column temperature was 60 ℃, the detection time was 15 min, the injection amount was 10 μL, the CAD atomizing chamber temperature was 35 ℃, and the filter was 5.0. Results: The recoveries of validation solutions at 50%, 80%, 100%, 120% and 150% concentrations were in the range of 90.8%-105.5%. The RSDs of peak areas were all below 10%. The RSD of measured value of validation solution at 100% concentration level was 2.7% after 6 times injection. Three batches of samples were prepared, each batch was prepared three times in parallel, and the mean values of measured results were 1.054, 1.041 and 0.995 mg·mL-1, with RSDs of 3.4%, 3.1% and 6.1%, respectively. Conclusion: This method is simple, rapid and sensitive, and can be used to accurately determine the trehalose content in monoclonal antibodies.
YU Xiao-juan, YU Chuan-fei, WANG Lan
. Development of an HPLC-CAD method to determine trehalose in therapeutic monoclonal antibody*[J]. Chinese Journal of Pharmaceutical Analysis, 2021
, 41(10)
: 1729
-1733
.
DOI: 10.16155/j.0254-1793.2021.10.09
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