Capillary zone electrophoresis ultraviolet (CZE‐UV) has become increasingly popular for the charge heterogeneity determination of mAbs and vaccines. The ε‐aminocaproic acid (eACA) CZE‐UV method has been used as a rapid platform method. However, in the last years, several issues have been observed, for example, loss in electrophoretic resolution or baseline drifts. Evaluating the role of eACA on the reported issues, various laboratories were requested to provide their routinely used eACA CZE‐UV methods, and background electrolyte compositions. Although every laboratory claimed to use the He et al. eACA CZE‐UV method, most methods actually deviate from He's. Subsequently, a detailed interlaboratory study was designed wherein two commercially available mAbs (Waters’ Mass Check Standard mAb [pI 7] and NISTmAb [pI 9]) were provided to each laboratory, along with two detailed eACA CZE‐UV protocols for a short‐end, high‐speed, and a long‐end, high‐resolution method. Ten laboratories participated each using their own instruments, and commodities, showing excellence method performance (relative standard deviations [RSDs] of percent time‐corrected main peak areas from 0.2% to 1.9%, and RSDs of migration times from 0.7% to 1.8% [n = 50 per laboratory], analysis times in some cases as short as 2.5 min). This study clarified that eACA is not the main reason for the abovementioned variations.
A controlled-release formulation was developed using enteric-coated beads in a hard gelatin capsule shell. This formulation requires two-stage dissolution testing at low and neutral pH. Because of high toxicity and limited stability of the active pharmaceutical ingredient in aqueous solution at room temperature, it is preferable to limit the analyst's contact with the sample solutions during testing, and aliquots from the dissolution vessel need to be quickly filtered and refrigerated immediately after sampling. To meet these challenges, a semi-automated dissolution method was developed and validated. Dissolution samples are withdrawn and filtered into vials in the HPLC sample compartment where the temperature is controlled at 4 °C. A fast HPLC method was developed with a 3-min run per injection, enabling analysis of the samples within a very short time after collection, thereby decreasing the sample storage time and potential for degradation. An evaluation was also conducted for comparison of serial and parallel operation of two dissolution baths for the two-stage testing. Serial operation was chosen for this method and applied with a medium-exchange technique, which is safe, robust, easy to perform, compliant with USP, and gives enhanced productivity.
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