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unige:91132 
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The importance of system band broadening in modern size exclusion chromatography |
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| Authors | ||
| Published in | Journal of Pharmaceutical and Biomedical Analysis. 2017, vol. 135, p. 50-60 | |
| Abstract | In the last few years, highly efficient UHP-SEC columns packed with sub–3 m particles were commercial-ized by several providers. Besides the particle size reduction, the dimensions of modern SEC stationaryphases (150 × 4.6 mm) was also modified compared to regular SEC columns (300 × 6 or 300 × 8 mm).Because the analytes are excluded from the pores in SEC, the retention factors are very low, ranging from−1 < k < 0, resulting in very small column band variance. Therefore, the contribution of the system itselfto peak variance can become significant under UHP-SEC conditions.The goal of this study was to evaluate the loss of efficiency observed with three different instruments(regular HPLC, non-optimized UHPLC and fully optimized UHPLC) offering different system variances. Itappears that the new 150 × 4.6 mm, sub–3 m SEC columns cannot be employed on a regular HPLCinstrument, since the efficiency loss was equal to 60–85%, when analyzing mAb sample. OptimizedUHPLC systems having very low extra-column volumes (typically Vec< 10 L) have therefore to be usedto properly operate these columns. Due to the instrument contribution to band broadening, the appar-ent efficiency of SEC columns packed with sub–2 m particles can indeed be hampered when usinginappropriate system. Considering the extra-column band broadening contribution of current UHPLCinstruments, a further decrease of SEC column dimension is therefore not desired. | |
| Keywords | Size exclusion chromatography — Kinetic efficiency — Extra-column band broadening — Aggregates — Monoclonal antibodies | |
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Article (Published version) (1.8 MB) - Limited access to UNIGE Other version: http://www.sciencedirect.com/science/article/pii/S073170851631069X |
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| Structures | ||
| Research group | Sciences analytiques | |
| Citation (ISO format) | GOYON, Alexandre, GUILLARME, Davy, FEKETE, Szabolcs. The importance of system band broadening in modern size exclusion chromatography. In: Journal of Pharmaceutical and Biomedical Analysis, 2017, vol. 135, p. 50-60. https://archive-ouverte.unige.ch/unige:91132 |
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