Optimizing Mobile Phase Composition, its Flow Rate and Column Temperature in HPLC Using an Experimental Design Assisted with A Simplex Method

1993 ◽  
Vol 16 (16) ◽  
pp. 3457-3470 ◽  
Author(s):  
Y. Guillaume ◽  
C. Guinchard
Keyword(s):  
Phase Composition ◽  
Experimental Design ◽  
Flow Rate ◽  
Simplex Method ◽  
Mobile Phase ◽  
Mobile Phase Composition ◽  
Column Temperature

scholarly journals A SIMPLE AND FAST RP-UHPLC-PDA METHOD FOR DETERMINATION OF ISRADIPINE FROM POLYMERIC NANOCAPSULES

2020 ◽  
pp. 225-229
Author(s):  
Fernanda Malaquias Barbosa ◽  
Guilherme dos Anjos Camargo ◽  
Amanda Martinez Lyra ◽  
Jessica Mendes Nadal ◽  
Paulo Vitor Farago
Keyword(s):  
High Performance Liquid Chromatography ◽  
Phase Composition ◽  
Flow Rate ◽  
Mobile Phase ◽  
High Performance ◽  
Experimental Basis ◽  
Mobile Phase Composition ◽  
Polymeric Systems ◽  
Polymeric Nanocapsules

A simple and fast analytical method of ultra-high performance liquid chromatography (UHPLC) was developed and validated in order to assay isradipine in poly(ε-caprolactone) (PCL)/polyethylene glycol (PEG) nanocapsules. Experiments were performed by UHPLC on a C18 chromatographic column at 25°C using a mobile phase composed by methanol and water (85:15 v/v) with a flow rate of 0.5 mL.min−1 and UV detection at 327nm for achieving a total run time of 1.5 min. The UHPLC method was validated according to the guidelines set on The International Conference on Harmonisation. It proved to be selective, linear (r = 0.99962), precise (RSD < 4.1%), and accurate (recovery rates between 95.24 and 96.53%) at the range from 10 to 40 µg.mL−1. The performance was robust when slight changes in the flow rate, wavelength of detection, and mobile phase composition were tested. It was successfully applied to quantify isradipine from nanoparticulate polymeric systems, showing high loading efficiency rates, greater than 98.55%. These results provided an experimental basis to use this method for quantifying isradipine with reliable results, besides being very fast, easy to perform and cheaper.

scholarly journals Revising Reverse-Phase Chromatographic Behavior for Efficient Differentiation of Both Positional and Geometrical Isomers of Dicaffeoylquinic Acids

2018 ◽  
Vol 2018 ◽  
pp. 1-11 ◽  
Author(s):  
Keabetswe Masike ◽  
Ian Dubery ◽  
Paul Steenkamp ◽  
Elize Smit ◽  
Edwin Madala
Keyword(s):  
Phase Composition ◽  
Uv Irradiation ◽  
Mobile Phase ◽  
Elution Order ◽  
Elution Profile ◽  
Plant Metabolites ◽  
Mobile Phase Composition ◽  
Reverse Phase ◽  
Geometrical Isomers ◽  
Column Temperature

Dicaffeoylquinic acids (diCQAs) are plant metabolites and undergo trans-cis-isomerization when exposed to UV irradiation. As such, diCQAs exist in both trans- and cis-configurations and amplify the already complex plant metabolome. However, analytical differentiation of these geometrical isomers using mass spectrometry (MS) approaches has proven to be extremely challenging. Exploring the chromatographic space to develop possible conditions that would aid in differentially separating and determining the elution order of these isomers is therefore imperative. In this study, simple chromatographic parameters, such as column chemistry (phenyl versus alkyl), mobile phase composition (methanol or acetonitrile), and column temperature, were investigated to aid in the separation of diCQA geometrical isomers. The high-performance liquid chromatography photodiode array (HPLC-PDA) chromatograms revealed four isomers post UV irradiation of diCQA authentic standards. The elution profile/order was seen to vary on different reverse-phase column chemistries (phenyl versus alkyl) using different mobile phase composition. Here, the elution profile/order on the phenyl-derived column matrices (with methanol as the mobile phase composition) was observed to be relatively reproducible as compared to the alkyl (C18) columns. Chromatographic resolution of diCQA geometrical isomers can be enhanced with an increase in column temperature. Lastly, the study highlights that chromatographic elution order/profile cannot be relied upon to fathom the complexity of isomeric plant metabolites.

Development and validation of high-performance liquid chromatography method for the simultaneous monitoring of pantoprazole sodium sesquihydrate and domperidone maleate in plasma and its application to pharmacokinetic study

2020 ◽  
Vol 32 (3) ◽  
pp. 157-165
Author(s):  
Ghulam Abbas ◽  
Malik Saadullah ◽  
Akhtar Rasul ◽  
Shahid Shah ◽  
Sajid Mehmood Khan ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Phase Composition ◽  
Flow Rate ◽  
Retention Time ◽  
Mobile Phase ◽  
High Performance ◽  
Mobile Phase Composition ◽  
Chromatography Method ◽  
Surface Design

A sensitive, inexpensive high-performance liquid chromatography–ultraviolet detection (HPLC–UV) method has been developed and validated for the simultaneous monitoring of pantoprazole sodium sesquihydrate (PSS) and domperidone maleate (DM) in rabbit plasma on a C18 column with UV detection at 285 nm. Box–Behnken design was used with 3 independent variables, namely, flow rate (X1), mobile phase composition (X2), and phosphate buffer pH (X3), which were used to design mathematical models. Response surface design was applied to optimize the dependent variables, i.e., retention time (Y1 and Y2) and percentage recoveries (Y3 and Y4) of PSS and DM. The method was sensitive and reproducible over 1.562 to 25 μg/mL. The effect of the quadratic outcome of flow rate, mobile phase composition, and buffer pH on retention time (p ˂ 0.001) and percentage recoveries of PSS and DM (p = 0.0016) were significant. The regression values obtained from analytical curve of PSS and DM were 0.999 and 0.9994, respectively. The percentage recoveries of PSS and DM were ranged from 94.5 to 100.41% and 94.77 to 100.31%, respectively. The developed method was applied for studying the pharmacokinetics of PSS and DM. The Cmax of test and reference formulations were 48.06 ± 0.347 μg/mL and 46.31 ± 0.398 μg/mL for PSS, and 15.11 ± 1.608 μg/mL and 12.06 ± 1.234 μg/mL for DM, respectively.

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