Modelling retention in liquid chromatography as a function of solvent composition and pH of the mobile phase

Bergés, Rosa; Sanz‐Nebot, Victoria; Barbosa, J.

doi:10.1016/s0021-9673(99)00915-2

Cited by 88 publications

(87 citation statements)

References 37 publications

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“…Due to the low content of methanol in the mobile phase, it was assumed that addition of MeOH to the buffer does not change pH of the hydroorganic mixture substantially. [31][32] From Figure 1 it can be seen that retention of uric acid, guanosine and adenine is markedly affected by pH. At pH 5.5 peaks are quite symmetrical and narrow but the chromatographic resolution (R) was poor, and at pH 4.0 it improves substantially and all bases were separated on the base line (R > 3.6).…”

Section: Optimization Of Chromatographic Conditions For Separationmentioning

confidence: 99%

Optimization of High Performance Liquid Chromatography Method for Simultaneous Determination of Some Purine and Pyrimidine Bases

Markelj

Župančič

Pihlar

2016

ACSi

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Nine purine and pyrimidine bases were separated and determined simultaneously using reversed phase (RP) high performance liquid chromatography (HPLC) in some food samples and biological fluids. Chromatographic behavior of these ionizable compounds highly depends on the interactions with the solvent as confirmed experimentally and by calculation of distribution of this species as a function of pH. Chromatograms show the optimal separation of five purine (uric acid, hypoxanthine, xanthine, adenine, and guanosine), and four pyrimidine (cytosine, uracil, cytidine and tymine) bases at pH around four. Accordingly, acetate buffer was selected due to high buffer capacity in this region. By variation of pH, concentration of buffer and volume ratio between buffer and methanol, we found that a mixture of 50 mM acetate buffer of pH 4.0 ± 0.1 with 3% of methanol ensures reproducibility, complete separation in less than 15 minutes and compatibility with UV and MS detection. Developed screening method was validated and applied for the analysis of complex clinical and beverage samples.

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Section: Optimization Of Chromatographic Conditions For Separationmentioning

confidence: 99%

Optimization of High Performance Liquid Chromatography Method for Simultaneous Determination of Some Purine and Pyrimidine Bases

Markelj

Župančič

Pihlar

2016

ACSi

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“…There are several approaches to predict the retention behavior of an ionisable solute as a function of chromatographic conditions. Because of the ionization properties of these types of solutes, the factors generally selected to optimize the chromatographic separation are the pH of the mobile phase and the percentage of the organic solvent of the eluent [3][4][5][6][7][8][9]. The effect of these parameters on the chromatographic behavior is somewhat more complex since the variation of organic modifier and analyze these compounds.…”

Section: Introductionmentioning

confidence: 99%

Identification of Penicillinase-Resistant Penicillins in Human Urine by RPLC Method

Üstün¹,

Demıralay²,

Nane³

2016

Eurasian J Anal Chem

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In this study, the combined effect of the organic modifier content and mobile phase pH has been used to predict the chromatographic behavior of methicillin, oxacillin, nafcillin, cloxacilin and dicloxacillin. The optimum separation condition of these drugs in acetonitrilewater binary mixture was determined from the mobile phase pH dependence of their retention factor. A Synergy Fusion-RP C18 analytical column (150 mm x 4.6 mm I.D., 4 µm) was preferred to carry out the developed method. Chromatographic measurements were done at 30• C with an eluent flow rate of 1 mL min -1 . In order to validate the optimized conditions, these drugs were applied successfully in spiked samples of human urine. Valsartan was chosen as an internal standard. The described method was linear over a concentration range of 2-32 µg mL -1 and 3-48 µg mL -1 for the assay of methicillin, nafcillin, dicloxacillin and oxacillin, cloxacillin respectively. Good linear relationships over the investigated concentration ranges were observed with values of correlation coefficient higher than 0.999 for all of the drugs. The intra-day and inter-day precisions of this method were evaluated with RSD (%) values less than 0,561 and 1,622%, respectively. Optimized assay method was validated according to United States Pharmacopeia (USP) guidelines to confirm linearity, accuracy and precision.

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“…There are several approaches to predict the retention behavior of an ionisable solute as a function of chromatographic conditions. Because of the ionization properties of these types of solutes, the factors generally selected to optimize the chromatographic separation are the pH of the mobile phase and the percentage of the organic solvent of the eluent [4][5][6][7][8][9][10][11][12][13][14]. The effect of these parameters on the chromatographic behavior is somewhat more complex since the variation of organic modifier concentration of the mobile phase induces a variation of the degree of ionization as well.…”

Section: Introductionmentioning

confidence: 99%

“…Equations that relate the retention of a weak acid compound in an LC column with the pH of the mobile phase consider that the retention factor is weighted average of the retention factor of the neutral and ionic forms of the compound according to the molar fractions of these forms in the eluent. A general model using activity coefficient for the influence on ionization on retention of weak acids is indicated below [5,7,10]:…”

Section: Introductionmentioning

confidence: 99%

The Combined Effect of the Organic Modifier Content and pH of the Mobile Phase on the Chromatographic Behavior of Some Arylpropionic and Arylacetic Acids to Optimize Their Liquid Chromatographic Determinations

et al. 2012

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The equation that express the combined effect of the organic modifier content and mobile phase pH has been used to predict the chromatographic behavior of flurbiprofen, ibuprofen, diclofenic acid, ketoprofen, fenoprofen, naproxen, bezafibric acid and clofibric acid. Correlations between the retention factors of the solutes and pH of the mobile phase were determined from a set of 40 chromatograms for each compound at four different levels of acetonitrile composition (40, 45, 50 and 55 %, v/v), and at ten pH values in each acetonitrile-water mixtures. In this way, a complete description of the retention behavior of each solute in the space defined by the pH and organic modifier percentages variables was obtained. The optimized chromatographic condition was applied to quantitative determination of flurbiprofen, ibuprofen, and diclofenic acid in pharmaceutical dosage forms.

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Modelling retention in liquid chromatography as a function of solvent composition and pH of the mobile phase

Cited by 88 publications

References 37 publications

Optimization of High Performance Liquid Chromatography Method for Simultaneous Determination of Some Purine and Pyrimidine Bases

Optimization of High Performance Liquid Chromatography Method for Simultaneous Determination of Some Purine and Pyrimidine Bases

Identification of Penicillinase-Resistant Penicillins in Human Urine by RPLC Method

The Combined Effect of the Organic Modifier Content and pH of the Mobile Phase on the Chromatographic Behavior of Some Arylpropionic and Arylacetic Acids to Optimize Their Liquid Chromatographic Determinations

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