High‐throughput capillary electrophoresis frontal analysis method for the study of drug interactions with human serum albumin at near‐physiological conditions

Martı́nez-Pla, J.J.; Martínez-Gómez, María Amparo; Martı́n-Biosca, Yolanda; Sagrado, S.; Villanueva-Camañas, R.M.; Medina-Hernández, M.J.

doi:10.1002/elps.200406049

Cited by 33 publications

(31 citation statements)

References 43 publications

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“…Even though CE-FA methods have been developed for a number of drugs and other ligands a reoccurring subject has been the lack of sensitivity of the UV detection employed [16,83,91,92] which makes it difficult to obtain a complete binding isotherm for high-affinity HSA ligands. Efforts have been made to develop rapid frontal analysis methods suitable for sceening of drug-HSA binding by using short end injection [118] and pressure-assisted CE-FA [92]. In addition to HSA, CE-FA methods for studying interactions with the other major plasmaproteins a 1 -acid glycoprotein [85,[119][120][121][122] and lipoproteins [84,[86][87][88]123] have been reported.…”

Section: Interactions Involving Plasma Proteins and Other Proteinsmentioning

confidence: 99%

Bioanalytical interaction studies executed by preincubation affinity capillary electrophoresis

Østergaard

Heegaard

2006

Electrophoresis

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The versatility of CE is beneficial for the study of many types of molecular interactions, because different experimental designs can be made to suit the characteristics of a particular interaction. A very versatile starting point is the preequilibration type of affinity CE that has been used extensively for characterizing biomolecular interactions in the last 15 years. We review this field here and include a comprehensive overview of the existing preincubation ACE modes including their advantages and limitations as well as the methodological developments and applications within the bioanalytical field.

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Section: Interactions Involving Plasma Proteins and Other Proteinsmentioning

confidence: 99%

Bioanalytical interaction studies executed by preincubation affinity capillary electrophoresis

Østergaard

Heegaard

2006

Electrophoresis

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“…In Table 1, the structure, bibliographic pK a values and octanol-water partition coefficients are shown. Using the methodology proposed in [16], the compound-HSA binding percentage (estimated at physiological conditions) and the affinity binding constants for HSA of the compounds studied were obtained (results in Table 1). As it can be observed, all the compounds considered in this study are basic and they present medium-high hydrophobicity.…”

Section: Multivariate Optimization Of Experimental Conditions For Commentioning

confidence: 99%

Multivariate optimization approach for chiral resolution of drugs using human serum albumin in affinity electrokinetic chromatography-partial filling technique

et al. 2005

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The enantiomeric resolution of chiral compounds using HSA by means of affinity EKC (AEKC)-partial filling technique is the result of a delicate balance between different experimental variables such as protein concentration, running pH (background electrophoretic buffer, protein and compound solutions) and protein solution plug length. In this paper multivariate optimization approaches for chiral separation of four basic drugs (alprenolol, oxprenolol, promethazine and propranolol) using HSA as chiral selector in AEKC-partial filling technique are studied. The experimental conditions to achieve maximum resolution are optimized using the Box-Behnken experimental design. Partial least squares and pareto charts are used to analyse the main effects on the resolution. The experimental resolutions observed for all compounds studied in optimum conditions agree with the estimated values based on response surface models. The results obtained show that the range of experimental conditions that provided enantioresolution narrows as hydrophobicity of analytes decreases. This fact can be explained by assuming that hydrophobicity controls the interaction of basic compounds with HSA.

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“…In FA, a relatively large sample plug is injected into the CE capillary to give rise to a flat plateau, the height of which is proportional to the concentration of analyte under equilibrium with albumin. The technique has been successfully developed to study the interaction between drug and albumin [20,21] and provides a promising procedure to investigate the interaction between bilirubin and albumin as well as to determine the free bilirubin concentration in a bilirubin-albumin mixture without disturbing the existing equilibrium. We have successfully coupled FA with CE to detect free bilirubin under undisturbed equilibrium condition and to monitor the interaction between bilirubin and albumin [22][23][24].…”

Section: Introductionmentioning

confidence: 99%

Microchip capillary electrophoresis for frontal analysis of free bilirubin and study of its interaction with human serum albumin

Nie

Fung

2008

Electrophoresis

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To meet the need for bedside monitoring of free bilirubin for neonates under critical conditions, a microfluidic chip was fabricated and tested for its coupling with CE/frontal analysis (FA) to determine free bilirubin and study of its binding interaction with HSA, which regulated its concentration in plasma. The poly(methyl methacrylate) (PMMA) multichannel chip was fabricated by CO2 laser ablation and bonded with a fused-silica separation capillary for CE/FA separation with UV detection. The chip was designed to allow a complete assay of four electrophoretic runs using preconditioned channels to speed up the determination of free bilirubin and to deliver quick results for bedside monitoring. Under optimized conditions, the linear working range for free bilirubin was from 10 to 200 micromol with RSDs from 2.1 to 5.0% for n=3, and the LOD at 9 micromol for S/N=3. From a binding study between bilirubin and HSA under FA condition, the second binding constant for bilirubin-HSA was determined as 1.07x10(5) L/mol and the number of binding sites per HSA as 3.46. The results enabled the calculation of free bilirubin for jaundiced infants based on the clinically significant level of total bilirubin, producing a range of 118.3-119.4 micromol/L. The developed method is shown to meet the clinical requirement with additional margin of protection to detect the early rising level of free bilirubin prior to jaundice condition. The low-cost microchip CE/FA device is shown to produce quick results with high potential to deliver a suitable bed-side monitoring method for bilirubin management in neonates.

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High‐throughput capillary electrophoresis frontal analysis method for the study of drug interactions with human serum albumin at near‐physiological conditions

Cited by 33 publications

References 43 publications

Bioanalytical interaction studies executed by preincubation affinity capillary electrophoresis

Bioanalytical interaction studies executed by preincubation affinity capillary electrophoresis

Multivariate optimization approach for chiral resolution of drugs using human serum albumin in affinity electrokinetic chromatography-partial filling technique

Microchip capillary electrophoresis for frontal analysis of free bilirubin and study of its interaction with human serum albumin

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