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HILIC/ESI-MS separation of acidic and other lipid classes using hydride column

Autoři: Cífková Eva | Hájek Roman | Lísa Miroslav | Holčapek Michal

Rok: 2016

Druh publikace: ostatní - přednáška nebo poster

Strana od-do: nestránkováno

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Jazyk	Název	Abstrakt	Klíčová slova
eng	HILIC/ESI-MS separation of acidic and other lipid classes using hydride column	Lipids are considered as multifunctional molecules having many important roles in the cell biology. Their comprehensive analysis is a challenging task due to the extreme complexity of individual lipid classes varying in the structure, attached functional groups, fatty acyl chains, polarity, dissociation and ionization behavior. Our previously published hydrophilic interaction liquid chromatography (HILIC) coupled with mass spectrometry (MS) method did not provide satisfactory results for acidic lipid classes (phosphatidic acid, lysophosphatidic acid, phosphatidylserine and lysophosphatidylserine) due to very broad tailing peaks under MS compatible conditions. For this purpose, the main parameters have been reoptimized with the emphasis on peak shapes of acidic lipid classes, such asthe type of stationary phases used in HILIC, pH of the mobile phase, the type and concentration of mobile phase additives. In comparison with other lipid classes, acidic lipid classes have more dissociation forms depending on the pH value, which can cause very broad and tailing chromatographic peaks and reduce the sensitivity. On the basis of the theoretical dissociation equilibria and experimental results, the pH value of mobile phase and the type of stationary phase have the crucial influence on peak shapes of these acidic lipid classes. The best results were obtained on new hydride type column using the mobile phase containing acetonitrile and 40 mmol/L of aqueous ammonium formate and pH 4 adjusted by formic acid. Hydride column has free silanol groups Si-OH on the surface of common silica columns replaced by hydride Si-H groups, which eliminate a strong association with polar analytes and can result in the best separation of acidic lipid classes on this type of column.	HILIC, ESI-MS; acidic lipid; hydride column

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