Published:Journal of Chromatographic Science,
ISSN 0021-9665Volume
41, Number 5, May/June 2003, pp 234-240
Online Extraction and Determination of Octylglucoside by Reversed-Phase
High-Performance Liquid Chromatography with Evaporative Light-Scattering Detection
John V. Amari[1], Zhirui Lian[2], Trish Lowden[2], Uditha deAlwis[2],
and Philip Levesque[2]
[1]EMD Pharmaceuticals, Inc., 125 Hartwell Avenue, Lexington, MA 02124 and
[2]Shire Biologics, Inc., 30 Bearfoot Road, Northborough, MA 01531
A reversed-phase high-performance liquid chromatography method using evaporative
light-scattering detection is developed for the determination of residual octylglucoside
(OG) levels after a detergent exchange step for in-process samples of a vaccine
antigen. The reversed-phase column not only provides separation of the OG but
also functions as an extraction column to remove the vaccine antigen from the
sample, thereby eliminating off-line sample manipulations. In addition to column
selection, the mobile phase is optimized to enhance extraction and separation.
The vaccine antigen is irreversibly bound to the column, allowing nonprotein
components to interact with the column for separation and elution. The assay
is linear over the range of 0.00050–0.050% OG. Precision tested at 0.0010%
and 0.0050% OG is 2.9% and 7.2% relative standard deviation, respectively.
The limits of quantitation and detection are determined to be 0.00050 and 0.000125%
OG, respectively. Accuracy is determined to be 103 and 98%, based on spike
recoveries of 0.0010% and 0.0050% OG, respectively.
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