For the evaluation of the efficiency of in vitro systems and for standardization of commercial products the reliable fast analytical procedures are required. These are usually based on HPLC analysis. Regular HPLC separations are usually time and solvent consuming. The new achievements in analytical equipment allows to apply much faster technique UPLC for plant phytochemical analysis. The technology is quite new (developed in 2004) and in this respect there is no literature available. It was applied for separation of phenolics in the extracts of four plant species: basil (Ocimium basilicum), dandelion (Taraxacum officinale), soybean (Glycine max), mint (Mentha piperica) considered as a source of nutraceuticals researched under the project NUTRASNACK (E.C. F.P.6 contract No FOOD-CT-2005-023044). The Acquinity Ultra Performance Liquid Chromatograph (Waters) consisting of Binary Solvent Manager, Sample Manager, PDA detector and Empower Pro 2.0 software was used. The analyses were performed on an UPLC BEH C18 column (1.7mm, 50mm ´ 2.1mm) utilizing a gradient elution profile and a mobile phase consisting of 0,1% acetic acid in water and 40% AcN. The column was maintained at 50oC and at a flow rate was kept constant at 0.35 mL/min. The separation profiles obtained for four analysed species were of similar quality as the profiles obtained with HPLC. However, optimization of the separation conditions (water-acetonitrile gradient shape, column temperature) in UPLC allowed us to reduce separation time down to 5 min (basil, dandelion) and 6 min (mint and soybean); regular HPLC separation time was 50 min. The developed method simplified the analytical protocol and shortened the time of analysis just to few minutes. This an important achievement when big number of samples e.g. in vitro culture efficiency evaluation is necessary.
The ultra-performance liquid chromatography (UPLC) analysis of phenolics in four plant species
PISTELLI, LUISA;BERTOLI, ALESSANDRA;
2007-01-01
Abstract
For the evaluation of the efficiency of in vitro systems and for standardization of commercial products the reliable fast analytical procedures are required. These are usually based on HPLC analysis. Regular HPLC separations are usually time and solvent consuming. The new achievements in analytical equipment allows to apply much faster technique UPLC for plant phytochemical analysis. The technology is quite new (developed in 2004) and in this respect there is no literature available. It was applied for separation of phenolics in the extracts of four plant species: basil (Ocimium basilicum), dandelion (Taraxacum officinale), soybean (Glycine max), mint (Mentha piperica) considered as a source of nutraceuticals researched under the project NUTRASNACK (E.C. F.P.6 contract No FOOD-CT-2005-023044). The Acquinity Ultra Performance Liquid Chromatograph (Waters) consisting of Binary Solvent Manager, Sample Manager, PDA detector and Empower Pro 2.0 software was used. The analyses were performed on an UPLC BEH C18 column (1.7mm, 50mm ´ 2.1mm) utilizing a gradient elution profile and a mobile phase consisting of 0,1% acetic acid in water and 40% AcN. The column was maintained at 50oC and at a flow rate was kept constant at 0.35 mL/min. The separation profiles obtained for four analysed species were of similar quality as the profiles obtained with HPLC. However, optimization of the separation conditions (water-acetonitrile gradient shape, column temperature) in UPLC allowed us to reduce separation time down to 5 min (basil, dandelion) and 6 min (mint and soybean); regular HPLC separation time was 50 min. The developed method simplified the analytical protocol and shortened the time of analysis just to few minutes. This an important achievement when big number of samples e.g. in vitro culture efficiency evaluation is necessary.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
