Testing experimental designs in liquid chromatography (II): Influence of the design geometry on the prediction performance of retention models

2021 ◽  
pp. 462458
Author(s):  
A. Gisbert-Alonso ◽  
J.A. Navarro-Huerta ◽  
J.R. Torres-Lapasió ◽  
M.C. García-Alvarez-Coque
2017 ◽  
Vol 52 (2) ◽  
pp. 125-134
Author(s):  
Matías N Baldo ◽  
Emmanuel Angeli ◽  
Natalia C Gareis ◽  
Gabriel A Hunzicker ◽  
Marcelo C Murguía ◽  
...  

A relative bioavailability study (RBA) of two phenytoin (PHT) formulations was conducted in rabbits, in order to compare the results obtained from different matrices (plasma and blood from dried blood spot (DBS) sampling) and different experimental designs (classic and block). The method was developed by liquid chromatography tandem-mass spectrometry (LC-MS/MS) in plasma and blood samples. The different sample preparation techniques, plasma protein precipitation and DBS, were validated according to international requirements. The analytical method was validated with ranges 0.20–50.80 and 0.12–20.32 µg ml−1, r > 0.999 for plasma and blood, respectively. Accuracy and precision were within acceptance criteria for bioanalytical assay validation (< 15 for bias and CV% and < 20 for limit of quantification (LOQ)). PHT showed long-term stability, both for plasma and blood, and under refrigerated and room temperature conditions. Haematocrit values were measured during the validation process and RBA study. Finally, the pharmacokinetic parameters (Cmax, Tmax and AUC0–t) obtained from the RBA study were tested. Results were highly comparable for matrices and experimental designs. A matrix correlation higher than 0.975 and a ratio of (PHT blood) = 1.158 (PHT plasma) were obtained. The results obtained herein show that the use of classic experimental design and DBS sampling for animal pharmacokinetic studies should be encouraged as they could help to prevent the use of a large number of animals and also animal euthanasia. Finally, the combination of DBS sampling with LC-MS/MS technology showed to be an excellent tool not only for therapeutic drug monitoring but also for RBA studies.


2008 ◽  
Vol 10 (1) ◽  
pp. 52-56
Author(s):  
Wojciech Zapała

Modeling of the influence of the modifier concentration on the retention process in NP-HPLC The adsorption model for an accurate prediction of the analyte retention in the normal - phase liquid chromatography with a binary mobile phase has been proposed. This model was derived using a thermody-namically consistent modified competitive Langmuir isotherm. The performance of the proposed equation was compared with two retention models reported in literature. All models were verified for different NP-LC systems by use of three criteria: the sum of squared differences between the experimental and theoretical data, approximation of the standard deviation and the Fisher test.


Separations ◽  
2021 ◽  
Vol 8 (4) ◽  
pp. 54
Author(s):  
Ester Peris-García ◽  
María José Ruiz-Angel ◽  
Juan José Baeza-Baeza ◽  
María Celia García-Alvarez-Coque

Hydrophilic interaction liquid chromatography (HILIC) is able to separate from polar to highly polar solutes, using similar eluents to those in the reversed-phase mode (RPLC) and a polar stationary phase, where water is adsorbed onto its surface. It is widely accepted that multiple modes of interaction take place in the HILIC environment, which can be far more complex than the interactions in an RPLC column. The behaviour in HILIC should be adequately modelled to predict the retention with optimisation purposes and improve the understanding on retention mechanisms, as is the case for RPLC. In this work, the prediction performance of several retention models is studied for seven HILIC columns (underivatised silica, and silica containing diol, amino and sulfobetaine functional groups, together with three columns recently manufactured with neutral, anionic, and cationic character), using uracil and six polar nucleosides (adenosine, cytidine, guanosine, thymidine, uridine, and xanthosine) as probe compounds. The results in HILIC are compared with those that were offered by the elution of several polar sulphonamides and diuretics analysed with two C18 columns (Chromolith Speed ROD and Zorbax Eclipse XDB). It is shown that eight retention models, which only consider partitioning or both partitioning and adsorption, give similar good accuracy in predictions for both HILIC and RPLC columns. However, the study on the elution strength behaviour, at varying mobile phase composition, reveals similarities (or differences) between RPLC and HILIC columns of diverse nature. The particular behaviour for the HILIC and RPLC columns was also revealed when the retention, in both modes, was fitted to a model that describes the change in the elution strength with the modifier concentration.


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