Practical HPLC Method Development

Snyder, Lloyd R.; Kirkland, J. J.; Glajch, Joseph L.

doi:10.1002/9781118592014

Cited by 1,358 publications

(1,337 citation statements)

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“…Lijima et al also used this method to isolate an antifungal peptide(AFP) from the hemolymph of a Sarcophaga peregrine (flesh fly) larva (24). It should be noted that, in general, separation of proteins by RP-HPLC would normally denature proteins, but small proteins (< 20,000 Da) can often be separated without significant degradation (27). In this study, the fractions obtained with SPE containing the antifungal activity was further purified by RP-HPLC.…”

Section: Discussionmentioning

confidence: 99%

Purification and Molecular Identification of an Antifungal Peptide from the Hemolymph of Musca domestica (housefly)

Fu¹,

Wu²,

Guo³

2009

Cell Mol Immunol

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Section: Discussionmentioning

confidence: 99%

Purification and Molecular Identification of an Antifungal Peptide from the Hemolymph of Musca domestica (housefly)

Fu¹,

Wu²,

Guo³

2009

Cell Mol Immunol

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“…Separation is, in turn, defined by resolution, which is inversely proportional to peak width. 11 To estimate the potential impact of loading on resolution using only a single component, we measured peak width versus load and then plotted the ratio Width min /Width load versus sample load (where Width min is the half-height peak width at lowest load and Width load is the half-height peak width at the measured load). This plot approximates the anticipated falloff 14 in resolution of this compound from a similarly behaving compound as a function of load.…”

Section: Evaluation Of System Loadingmentioning

confidence: 99%

Evaluation of a New Preparative Supercritical Fluid Chromatography System for Compound Library Purification: The TharSFC SFC-MS Prep-100 System

Ebinger

Weller

Kiplinger³

et al. 2011

JALA: Journal of the Association for Laboratory Automation

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Preparative HPLC-MS is often the method of choice for purification of small amounts (<100 mg) of diverse new molecules, such as compound libraries for drug discovery. The method is robust, well proven, and widely applicable. In contrast, preparative supercritical fluid chromatography coupled with mass spectrometry (SFC-MS) has seen only slow acceptance for the same application—despite some potential scientific and economic advantages. One of the reasons for slow adoption of SFC-MS is the lack of well-proven, robust, and commercially available instrumentation. In early 2009, TharSFC (a Waters Company, Pittsburgh, PA) introduced a new fully integrated system for preparative SFC-MS: The SFC-MS Prep-100. We report herein an objective evaluation of the SFC-MS Prep-100, including tests for pump and autosampler performance, sample recovery, sample carryover, fraction triggering, detector/fraction collector synchronization, and overall robustness. Our results suggest that the SFC-MS Prep-100 represents a significant advance over previous generation instrumentation.

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“…Taking into account the baseline resolution (R s >1.5 for bands of similar size [21], only LIC8 column separated 11 PPs to baseline (R s values are in Table 5). Other columns also separated 11 PPs: LC18 and SF (R s = 1.5) for PCP and 24DMP, respectively; SM12 (R s = 1.4) for PCP, and HYP (R s = 1.3) for 4NP, exhibiting the remainder compounds R s A1.…”

Section: Stationary Phase Effectmentioning

confidence: 99%

“…The LC columns widely used are Spherisorb ODS-2 [2, 14 -17] and Hypersil Green [11, 18 -20]. These packings, which are mainly based on Type-A alkyl silica, are less pure and more acidic silica, resulting in an increased silanol ionization as a function of pH, and presumably increased hydrogen-bond acidity [21]. Little attention has been paid to Type-B alkyl silica columns (highly purified, less acidic silica) for PPs separation [22].…”

Section: Introductionmentioning

confidence: 99%

Comparison of the performance of different reversed‐phase columns for liquid chromatography separation of 11 pollutant phenols

Castro

Santos-Montes

Izquierdo-Hornillos³

et al. 2007

J of Separation Science

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A systematic optimization of the HPLC separation of a mixture containing 11 pollutant phenols (PPs) using a Hypersil ODS (250 mm64.6 mm, 5 lm) column and UV-DAD detection has been carried out. The binary mobile phases used were obtained by mixing 50 mM phosphate (pH = 3.0) and methanol, ACN, or THF as organic modifiers. After selecting ACN as an organic modifier, the effects of pH and temperature on PPs separation were studied. A mobile phase of 50 mM acetate (pH = 5.0) -ACN (60 : 40 v/v) at 508C allowed the separation of 11 phenols but not to baseline in 17 min. To improve the performance of this separation, the following RP columns were tested: Luna C18 (2), Purospher C18, Synergi C12, Synergi Fusion C18, Gemini C18, Luna Cyano, Lichrospher C8, and Envirosep-PP (polymeric). In all the cases, the performance (analysis time, retention, selectivity, resolution, asymmetry factors, and efficiency) was evaluated. A further reoptimization of the mobile phase was carried out for all the columns by studying the ACN content and pH, with the aim of improving the above-mentioned separations and selecting the most suitable one for PPs analysis.

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Practical HPLC Method Development

Cited by 1,358 publications

References 0 publications

Purification and Molecular Identification of an Antifungal Peptide from the Hemolymph of Musca domestica (housefly)

Purification and Molecular Identification of an Antifungal Peptide from the Hemolymph of Musca domestica (housefly)

Evaluation of a New Preparative Supercritical Fluid Chromatography System for Compound Library Purification: The TharSFC SFC-MS Prep-100 System

Comparison of the performance of different reversed‐phase columns for liquid chromatography separation of 11 pollutant phenols

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