Some solutions to obtain very efficient separations in isocratic and gradient modes using small particles size and ultra-high pressure

Guillarme, Davy; Grata, Elia; Glauser, Gaëtan; Wolfender, Jean‐Luc; Veuthey, Jean‐Luc; Rudaz, Serge

doi:10.1016/j.chroma.2009.02.032

Cited by 66 publications

(28 citation statements)

References 56 publications

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“…3C and B, which was equal to 20%, was quite limited in proportion to the increase in analysis time (4-fold) and to the generated backpressure. This observation is in good agreement with recent UHPLC studies [39][40][41] that show the longest column does not necessarily provide an important increase in gradient performance as the latter depends on both isocratic efficiency and the column dead time, which is elevated with longer columns operating at low flow rate.…”

Section: Ultra-fast and Highly Efficient Separations Of Catechinssupporting

confidence: 80%

High throughput qualitative analysis of polyphenols in tea samples by ultra-high pressure liquid chromatography coupled to UV and mass spectrometry detectors

Guillarme

Casetta

Bicchi

et al. 2010

Journal of Chromatography A

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a b s t r a c tThe analysis of polyphenols in tea extracts is important due to their potential health benefits. Therefore, efficient and high throughput analytical methods have been developed for the separation of seven predominant polyphenols, also known as catechin derivatives, present in tea extracts. Columns packed with sub-2-m particles operating at elevated pressure (UHPLC strategy) were selected to improve chromatographic performance. The potential of UHPLC-UV was demonstrated with baseline resolution of all standard catechins in only 30 s using a 50-mm column packed with 1.7-m particles. When dealing with real samples such as tea extracts, however, longer columns of up to 150 mm in length were employed to enhance the separation of catechin derivatives and other constituents within the tea samples while maintaining an acceptable analysis time. Two strategies based on 2-D experiments were proposed to clearly identify catechins. Firstly, a liquid-liquid extraction procedure was added prior to the UHPLC-UV analysis to decrease the complexity of the sample. Secondly, UHPLC was coupled to ESI-MS/MS to attain sufficient sensitivity and selectivity between catechin derivatives and other constituents of tea extract. These two strategies were found extremely promising as a clear discrimination of catechins from the matrix could be attained.

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Section: Ultra-fast and Highly Efficient Separations Of Catechinssupporting

confidence: 80%

High throughput qualitative analysis of polyphenols in tea samples by ultra-high pressure liquid chromatography coupled to UV and mass spectrometry detectors

Guillarme

Casetta

Bicchi

et al. 2010

Journal of Chromatography A

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“…To this end, various techniques have been introduced, including the use of sub-2 μm particles, high temperature, monolithic columns and superfi cially porous stationary phases (Carr et al, 2009;Zhang et al, 2009;Poppe, 1997;Plumb et al, 2007;McNeff et al, 2007;Fekete et al, 2010;Guillarme et al, 2009). The term UHPLC, which stands for ultra high-performance liquid chromatography, may be used to indicate any one of these techniques.…”

Section: Chromatographymentioning

confidence: 99%

Systematic LC‐MS/MS bioanalytical method development that incorporates plasma phospholipids risk avoidance, usage of incurred sample and well thought‐out chromatography

Jemal

Ouyang

Xia

2009

Biomedical Chromatography

184

124

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This treatise summarizes the underlying principle and the road map for systematic LC-MS/MS bioanalytical method development. The three themes that have recently emerged as central to building quality during method development-phospholipids, incurred sample and sound chromatographic considerations-are the main focus of this article. In order to reduce the bioanalytical risk associated with plasma phospholipids, a dual approach involving extraction and chromatography is recommended. The use of incurred sample during method development is essential to avoid interference arising from drug-related components. This is different from the current practice of incurred sample reanalysis, which tests reproducibility during method application. LC column/mobile phase optimization is needed to achieve appropriate peak shape, sensitivity and the separation of the analyte from interfering metabolites and phospholipids. Related to sound chromatographic considerations, we lay out facts and myths related to UPLC, vis-à-vis HPLC. Incorporation of quality during method development avoids the costly experience of finding out by chance about the invalidity of a method after it has been used in support of a number of pivotal clinical and non-clinical studies. To this end, we put forth an outline of a protocol for a systematic LC-MS/MS bioanalytical method development.

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“…Second, small particles can be used to achieve high resolution by using longer columns operated at flow rates close to the optimum. Literature provides examples of the use of UHPLC with sub-2 lm columns at high temperature to decrease the analysis time [9][10][11] or to enhance the efficiency [12]. Coupling several analytical columns together is a simple way to achieve high efficiency in HPLC, but the challenge is how to implement this approach using commercially available instrumentation.…”

Section: Introductionmentioning

confidence: 99%

“…Monolithic material has significant drawbacks, such as the limited resistance of silica-based monoliths to elevated pressure, temperature and pH. In addition, a limited number of column chemistries and dimensions are available [12].…”

Section: Introductionmentioning

confidence: 99%

High-Efficiency Liquid Chromatography Using Sub-2 μm Columns at Elevated Temperature for the Analysis of Sulfonamides in Wastewater

Shaaban

Górecki

2011

Chromatographia

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Efficiency in HPLC can be enhanced by increasing the column length and/or decreasing the particle size. The use of high temperature in HPLC has emerged as a valuable tool to overcome the increase in column backpressure when using small packing particles, as it allows for reduction in mobile phase viscosity. In this study, high plate count was obtained by coupling sub-2 lm columns at elevated temperature to reduce the viscosity of the mobile phase, thus reducing the column backpressure. At 80°C, up to three columns of 15 cm 9 4.6 mm I.D. packed with 1.8 lm particles could be coupled generating *84,000 theoretical plates for the last eluting compound. The number of theoretical plates was increased on average by a factor of *3.6 when three columns were coupled at 80°C compared with one column at 30°C. The relationships between separation efficiency and column length were examined using Van Deemter plots constructed at 30°C and 80°C for different column lengths. The advantages of using coupled columns in combination with elevated temperature for the environmental analysis were illustrated using test mixtures comprised of eight sulfonamides separated on one column at 30°C and three coupled columns at 80°C by isocratic elution. Sample clean up was carried out by employing solid-phase extraction (SPE) using Oasis HLB cartridges. The method developed was validated based on parameters such as linearity, precision, accuracy, detection and quantification limits. Recoveries generally ranged from 71.7 to 99% (with the exception of sulfanilamide), with standard deviations not higher than 4.7%. The detection limits of the method ranged from 0.6-2 lg L -1 , while limits of quantification were in the range 2-6.7 lg L -1 with UV detection.

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Some solutions to obtain very efficient separations in isocratic and gradient modes using small particles size and ultra-high pressure

Cited by 66 publications

References 56 publications

High throughput qualitative analysis of polyphenols in tea samples by ultra-high pressure liquid chromatography coupled to UV and mass spectrometry detectors

High throughput qualitative analysis of polyphenols in tea samples by ultra-high pressure liquid chromatography coupled to UV and mass spectrometry detectors

Systematic LC‐MS/MS bioanalytical method development that incorporates plasma phospholipids risk avoidance, usage of incurred sample and well thought‐out chromatography

High-Efficiency Liquid Chromatography Using Sub-2 μm Columns at Elevated Temperature for the Analysis of Sulfonamides in Wastewater

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