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Off‐line two‐dimensional liquid chromatography separation for the quality control of saponins samples from Quillaja Saponaria
Author(s) -
Lecas Lucile,
Nuccio Sylvie,
Vaumas René,
Faure Karine
Publication year - 2021
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.202100115
Subject(s) - chromatography , chemistry , hydrophilic interaction chromatography , high performance liquid chromatography , reversed phase chromatography , column chromatography , chromatography column
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 nonconform samples. To fulfill quality control constraints, off‐line configuration using basic instrumentation was promoted. Hence, reversed‐phase liquid chromatography × reversed‐phase liquid chromatography and hydrophilic interaction chromatography × reversed‐phase liquid chromatography methods with ultraviolet and single‐quadrupole mass spectrometry detection were kinetically optimized. The reversed‐phase liquid chromatography × reversed‐phase liquid chromatography method used a pH switch between dimensions to maximize orthogonality. Despite diagonalization, it led to a high peak capacity of 831 in 2 h. On the other hand, the combination of hydrophilic interaction chromatography and reversed‐phase liquid chromatography 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 liquid chromatography × reversed‐phase liquid chromatography contour plot highlighted the loss of fatty acid chains, while the hydrophilic interaction chromatography × reversed‐phase liquid chromatography method was found useful to evidence enzymatic loss of sugar moieties.