Betaine is an amino acid widely present in animalsand plants, such as beetroots, where polar betaine is accumulated in response to water stresses. Betaine takes part in the protection of cells, proteins and enzymes from environmental stress and participates in the methionine cycle in human liver and kidneys. The most important feature is however betaine biological relevance in reducing cancer development. Despite these biological effects of betaine, just few analytical methods for its extraction from highly accumulating beetrootsare by far reported; these approaches are mainly based on solid phase extraction (SPE) that however does not provide quantitative recoveries. The aim of this work was the development of a new, quick and effective procedure for the isolation and determination of betaine from two different varieties of Beta Vulgaris (Red and Gold). For the first time an accelerated solvent extraction (ASE) with methanol was coupled with solid phase extraction on silica cartridge to isolate betaine from interfering species coextracted. Additionally, the performance of a micro extraction by packed sorbent (MEPS) after the ASE extraction was also studied. A modified QuEChERS procedure was finally tested in order to join the extraction and clean-up stages in a unique analytical protocol. Qualitative and quantitative analyses were performed with the use of LC-MS/MS (negative ESI, MRM mode) after separation by hydrophilic interaction liquid chromatography (HILIC). The mass spectrometry parameters were optimized by means of a Central Composite Design. Recoveries of betaine from beetroots were about 97% (RSD<6%) with the use of ASE coupled to SPE. The total content of betaine in extracts of various part of plants (juice, peel, root) have been determined and compared.
Determination of biological active compounds in food matrices: comparison of sample pretreatment procedures for the extraction of betaine from Chenopodiacee roots and determination by HPLC-MS/MS
RIVOIRA, LUCA;BRUZZONITI, Maria Concetta
2016-01-01
Abstract
Betaine is an amino acid widely present in animalsand plants, such as beetroots, where polar betaine is accumulated in response to water stresses. Betaine takes part in the protection of cells, proteins and enzymes from environmental stress and participates in the methionine cycle in human liver and kidneys. The most important feature is however betaine biological relevance in reducing cancer development. Despite these biological effects of betaine, just few analytical methods for its extraction from highly accumulating beetrootsare by far reported; these approaches are mainly based on solid phase extraction (SPE) that however does not provide quantitative recoveries. The aim of this work was the development of a new, quick and effective procedure for the isolation and determination of betaine from two different varieties of Beta Vulgaris (Red and Gold). For the first time an accelerated solvent extraction (ASE) with methanol was coupled with solid phase extraction on silica cartridge to isolate betaine from interfering species coextracted. Additionally, the performance of a micro extraction by packed sorbent (MEPS) after the ASE extraction was also studied. A modified QuEChERS procedure was finally tested in order to join the extraction and clean-up stages in a unique analytical protocol. Qualitative and quantitative analyses were performed with the use of LC-MS/MS (negative ESI, MRM mode) after separation by hydrophilic interaction liquid chromatography (HILIC). The mass spectrometry parameters were optimized by means of a Central Composite Design. Recoveries of betaine from beetroots were about 97% (RSD<6%) with the use of ASE coupled to SPE. The total content of betaine in extracts of various part of plants (juice, peel, root) have been determined and compared.
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