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Separation of somatropin charge variants by multiple‐injection CZE with P olybrene/chondroitin sulfate A double‐coated capillaries
Author(s) -
Amini Ahmad
Publication year - 2013
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201300327
Subject(s) - chemistry , chondroitin sulfate , chromatography , capillary electrophoresis , electrophoresis , capillary action , coating , dermatan sulfate , analytical chemistry (journal) , glycosaminoglycan , materials science , biochemistry , organic chemistry , composite material
The performance of dynamic double‐coated fused‐silica capillaries with P olybrene and chondroitin sulfate A has been compared with uncoated fused‐silica capillaries for the determination of recombinant human growth factor (somatropin) charge variants. The separations were carried out under the same electrophoretic conditions as described in the E uropean P harmacopoeia, i.e. at pH 6.0 and 30°C. The coating significantly reduced the interactions between the proteins and the surface of the fused‐silica capillary. The first five separations performed in a new bare fused‐silica capillary were discarded because of very poor separation performance as a result of protein–surface interactions. There was an approximate twofold increase in the interday migration time precision (% RSD ≤ 6.5%) in the double‐coated capillaries. The method was successfully transferred to a multiple CZE mode where two samples were analyzed in a single electrophoretic run. The average purity of somatropin certified reference standard was 98.0% (% RSD ≤ 0.3%) determined by using uncoated and coated capillaries.