LAUSR

Quil-A is a purified extract of saponins with strong immunoadjuvant activity. While specific molecules have been identified and tested in clinical trials, Quil-A is mostly used as a totum of the Quillaja Saponaria bark extract. Quality control of the extract stability is usually based on the monitoring of specific saponins, whereas the comparison of samples with an initial chromatogram seems more appropriate. A reference fingerprint based on comprehensive two-dimensional liquid chromatography offers a rapid detection of non-conform samples. To fulfill quantity control constraints, off-line configuration using basic instrumentation was promoted. Hence, reversed-phase LC × reversed-phase LC and hydrophilic interaction chromatography × reversed-phase LC methods with UV and single-quadrupole MS detection were kinetically optimized. The reversed-phase LC × reversed-phase LC method used a pH switch between dimensions to maximize orthogonality. Despite diagonalization, it led to a high peak capacity of 831 in two hours. On the other hand, the combination of hydrophilic interaction chromatography and reversed-phase LC offered a larger orthogonality but a lower, yet satisfactory peak capacity of 673. The advantages of both methods were illustrated on degraded samples, where the reversed-phase LC × reversed-phase LC contour plot highlighted the loss of fatty-acid chains, while the hydrophilic interaction chromatography × reversed-phase LC method was found useful to evidence enzymatic loss of sugar moieties. This article is protected by copyright. All rights reserved.