Objective: To establish and optimize the method for determining the purity and potency of asparaginase(Escherichia coli) and asparaginase(Erwinia carotovora) and improve its quality standards. Methods: The separation effects of Xbridge protein BEH SEC column (300 mm×7.8 mm, 3.5 μm, 200Å) and TSK G3000swxl (300 mm×7.8 mm, 5 μm) were tested and the mobile phase was optimized for the purity method. The form of the dosage-effect relationship and the linear range of assay were examined, and meanwhile the feasibility of the slope ratio model and the standard curve model were evaluated. Results: Improved resolution between the impurity peak and the main peak were obtained using the Xbridge protein BEH SEC column. Besides, the assay results showed that, it was a linear response for both of the asparaginases in the dosage range of 2-30 U·mL-1 and the regression coefficient r could be greater than 0.995. Meanwhile,the standard curve measurement method was more accurate and simple, the sample using double dosage could effectively eliminate the experimental errors, and the measurement deviation could be less than ±2.0% between two dosages. Conclusion: Compared with the absolute method, the relative method has greatly improved the accuracy and reproducibility of potency determination. Better resolutions can be obtained by using the optimized purity determination method, which can better avoid the result error caused by the integration method.
WANG Yue, CHEN Xin-tong, LI Jing, FAN Hui-hong
. Study on the methods of purity and potency of asparaginase*[J]. Chinese Journal of Pharmaceutical Analysis, 2022
, 42(1)
: 156
-165
.
DOI: 10.16155/j.0254-1793.2022.01.18
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