Performance of different C18 columns in reversed-phase liquid chromatography with hydro-organic and micellar-organic mobile phases

Ruiz-Ángel, M.J.; Pous-Torres, S.; Carda‐Broch, Samuel; Garcı́a-Alvarez-Coque, M.C.

doi:10.1016/j.chroma.2014.04.011

Cited by 23 publications

(13 citation statements)

References 24 publications

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“…This result agrees with Equation (14). Finally, representing A × F 2 versus the flow rate ( Figure 5c), a parabola is also obtained, where the effect of high flow rates is highlighted.…”

Section: Optimal Flow Ratesupporting

confidence: 88%

“…On the other hand, this approach is based on the theoretical plate model, which assumes Gaussian peaks for sufficiently large efficiencies [10]. However, peaks are usually tailing [11][12][13][14], and in some cases, they are fronting [15]. Even in these cases, Equation (3) is applied, using numerical integration to obtain the retention times and variances.…”

Section: Introductionmentioning

confidence: 99%

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Prediction of Peak Shape and Characterization of Column Performance in Liquid Chromatography as a Function of Flow Rate

Baeza-Baeza

Ortiz-Bolsico

Garcı́a-Alvarez-Coque

2015

Chromatography

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Traditionally, column performance in liquid chromatography has been studied using information from the elution of probe compounds at different flow rates through van Deemter plots, which relate the column plate height to the linear mobile phase velocity. A more recent approach to characterize columns is the representation of the peak widths (or the right and left peak half-widths) for a set of compounds versus their retention times, which, for isocratic elution, give rise to almost linear plots. In previous work, these plots have been shown to facilitate the prediction of peak profiles (width and asymmetry) with optimization purposes. In this work, a detailed study on the dependence of the peak widths (or half-widths) on the flow rate is reported. A new approach to quantify the deterioration of column performance for slow and fast flow rates and to characterize chromatographic columns is proposed. The approach makes use of the width (or half-widths) for a set of compounds with similar interaction kinetics and does not require knowledge of the extra-column contributions to the total variance. The chromatographic data of two sets of compounds of different natures (sulfonamides and β-blockers), eluted from Spherisorb and Chromolith columns with acetonitrile-water mixtures, are used to illustrate the approach.

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“…This result agrees with Equation (14). Finally, representing A × F 2 versus the flow rate ( Figure 5c), a parabola is also obtained, where the effect of high flow rates is highlighted.…”

Section: Optimal Flow Ratesupporting

confidence: 88%

Section: Introductionmentioning

confidence: 99%

Prediction of Peak Shape and Characterization of Column Performance in Liquid Chromatography as a Function of Flow Rate

Baeza-Baeza

Ortiz-Bolsico

Garcı́a-Alvarez-Coque

2015

Chromatography

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“…Many types of columns have been used for separation and retention of PCs . The most widely used have been silica‐based C 18 columns.…”

Section: Resultsmentioning

confidence: 99%

Robust method for the analysis of phytochelatins in rice by high-performance liquid chromatography coupled with electrospray tandem mass spectrometry based on polymeric column materials

Bian

Zhou

et al. 2015

J. Sep. Science

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A sensitive and robust high-performance liquid chromatography coupled with electrospray tandem mass spectrometry method for the identification and quantification of glutathione and phytochelatins from rice was developed. Homogenized samples were extracted with water containing 100 mM dithiothreitol, and solid-phase extraction using polymer anion exchange resin was employed for sample purification. Chromatography was performed on a polymeric column with acetonitrile and water containing 0.1% formic acid as the mobile phase at the flow rate of 300 μL/min. The limit of quantitation was 6-100 nM. This assay showed excellent linearity for both glutathione and phytochelatins over physiological normal ranges, with correlation coefficients (r) > 0.9976. Recoveries for four biothiols were within the range of 76-118%, within relative standard deviations less than 15%. The intraday precision (n = 7) was 2.1-13.3%, and the interday precision over 15 days was 4.3-15.2%. The optimized method was applied to analyze tissue samples from rice grown using nutrient solutions with three different cadmium concentrations (0, 50, and 100 μM). With increasing cadmium concentrations, the content of phytochelatin 2 and phytochelatin 3 in rice roots increased, in contrast to most phytochelatins, and the content of glutathione in rice stems and roots decreased significantly.

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“…Genistein, daidzein, and glycitein can be separated by reversed-phase HPLC using the C18 column [11][12][38][39] due to the possibility of interaction between isoflavone aglycones as organic compounds toward the silica-based column [40]. Methanol was chosen as one of the mobile phase compositions since it was reported that aglycone isoflavone such as genistein was played an important role as either a hydrogen bond donor or acceptor to interact with methanol and resulted in extra stability for the whole system [41].…”

Section: Rsm Optimization Of Genistein Daidzein and Glycitein Separmentioning

confidence: 99%

Employing an R Software Package rsm for Optimizing of Genistein, Daidzein, and Glycitein Separation and Its Application for Soy Milk Analysis by HPLC Method

et al. 2020

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Soy milk, one of the soybean products, become more popular in recent years due to its benefit for human health. Biological activities of soybean products have been widely studied according to the presence of isoflavone aglycones content, including genistein, daidzein, and glycitein. Hence, it is important to develop an effective and efficient analytical method to provide guidance regarding appropriate isoflavone intake levels for soy milk. A reversed-phase high performance liquid chromatography (HPLC) method was developed and optimized in this study employed by R statistical software with the package of rsm. A C18 column was used for HPLC separation with the detection at 260 nm. Optimized condition for HPLC separation has been achieved with the mobile phase of methanol: water (63.26:36.74), a flow rate of 0.81 mL/min, and a column temperature of 45.31 °C. These conditions were applied in the HPLC system and successfully tested for the system suitability. Quantitative estimation was performed and resulted that the genistein, daidzein, and glycitein content in soy milk samples were 6.372, 6.273, and 2.853 µg/mL, respectively.

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Performance of different C18 columns in reversed-phase liquid chromatography with hydro-organic and micellar-organic mobile phases

Cited by 23 publications

References 24 publications

Prediction of Peak Shape and Characterization of Column Performance in Liquid Chromatography as a Function of Flow Rate

Prediction of Peak Shape and Characterization of Column Performance in Liquid Chromatography as a Function of Flow Rate

Robust method for the analysis of phytochelatins in rice by high-performance liquid chromatography coupled with electrospray tandem mass spectrometry based on polymeric column materials

Employing an R Software Package rsm for Optimizing of Genistein, Daidzein, and Glycitein Separation and Its Application for Soy Milk Analysis by HPLC Method

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