Miniaturisation of solid phase extraction method for determination of retinol, alpha- and gamma-tocopherol in human serum using new technologies

Krčmová, Lenka Kujovská; Solichová, Dagmar; Plíšek, Jiří; Kašparová, Markéta; Sobótka, L.; Solich, Petr

doi:10.1080/03067310903267299

Cited by 11 publications

(2 citation statements)

References 21 publications

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“…L is the column length. At 10% height of the pulse chromatogram peak, a is the front half‐width, b is the back half‐width [19]. The dimensionless Peclet number ( Pe ) demonstrating the overall axial dispersion of fabrics as one of the indicators for hydrodynamic behavior was calculated according to Ref.…”

Section: Methodsmentioning

confidence: 99%

Megaporous monolithic adsorbents for bioproduct recovery as prepared on the basis of nonwoven fabrics

et al. 2022

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Megaporous adsorbents were prepared based on nonwoven polyethylene terephthalate (PET) fabrics and functionalized by covalent modification with polyvinylamine (PVAm) or monotriazinyl-β-cyclodextrin-substituted polyvinylamine (PVAm-MCT-β-CD). Mechanical properties of the resulting fabrics were maintained, as judged by tensile strength tests and scanning electron microscopy. Exceptional porosity (≥82%) and preserved hydrodynamic characteristics (Pe ≥ 63) indicated excellent structural stability when packed. The performance of the constructed adsorbents was evaluated with high molecular weight (proteins) and low molecular weight (dyes) model compounds. The static binding capacity (SBC) for bovine serum albumin (BSA) was 79.7 ± 1.3 and 92.9 ± 8.2 mg/g for PVAm-modified and PVAm-MCT-β-CD-modified fabrics, respectively. The mentioned materials also adsorbed Orange II, an acidic dye (92.4 ± 2.6 and 101.9 ± 2.6 mg/g, respectively), indicating that the hydrophobicity was a prevailing binding mechanism operating at a pH close to isoelectric point. SBC for lysozyme and toluidine blue O (TBO, a basic dye) onto PVAm-MCT-β-CD functionalized PET was 52.7 ± 5.1 and 73.3 ± 0.6 mg/g, respectively. TBOs have also shown some affinity for PVAm functionalized PET, but this was most likely to be mediated by hydrophobicity. On the other hand, operating at a superficial velocity of 90 cm/h, dynamic binding capacity for BSA was 11.4 ± 3.5 and 2.5 ± 0.6 mg/g indicating the importance of possible aggregation mechanisms during protein binding at equilibrium. Thus, PET-based adsorbents require further functional improvement for chromatography applications. However, the

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Section: Methodsmentioning

confidence: 99%

Megaporous monolithic adsorbents for bioproduct recovery as prepared on the basis of nonwoven fabrics

et al. 2022

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“…This method usually involves extraction with an organic solvent, centrifugation for phase separation, organic layer evaporation, and residue reconstitution in the appropriate solvent. Since this procedure is relatively tedious and time consuming, other techniques such as simple protein precipitation , miniaturized SPE , and SPE with electrospun nanofibers have been used. The addition of antioxidants and saponification step are common when complex matrices are extracted.…”

Section: Introductionmentioning

confidence: 99%

Rapid determination of lipophilic vitamins in human serum by ultra‐high performance liquid chromatography using a fluorinated column and high‐throughput miniaturized liquid–liquid extraction

Červinková

Krcmova

Klabackova

et al. 2017

J of Separation Science

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A high-throughput miniaturized liquid-liquid extraction procedure followed by a simple ultra-high performance liquid chromatography method coupled with fluorescence detection for bioanalytical analysis of all tocopherol isomers and retinol in human serum has been developed and validated. In the extraction procedure, a synthetic internal standard tocol was used, which does not occur in the human body. The separation of structurally related vitamins was achieved using a new generation of pentafluorophenyl propyl core-shell stationary phase with elution using methanol and an aqueous solution of ammonium acetate. The fluorescence of retinol and tocopherol isomers was detected at λ = 325, 295 nm and λ = 480, 325 nm, respectively. The rapid baseline separation of all analytes was accomplished within 4.0 min. The sensitivity of method was demonstrated with lower limits of quantification: retinol 0.01 μM, α-tocopherol 0.38 μM, β-tocopherol 0.18 μM, γ-tocopherol 0.14 μM, and δ-tocopherol 0.01 μM. Possible application of this method in clinical practice was confirmed by the analysis of human serum samples from healthy volunteers. Finally, the simultaneous determination of retinol and all tocopherol isomers in human serum can enable the clarification of their role in metabolism and in diseases such as cancer.

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Comparison of a new high-resolution monolithic column with core-shell and fully porous columns for the analysis of retinol and α-tocopherol in human serum and breast milk by ultra-high-performance liquid chromatography†

et al. 2013

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The reduction of analysis time, cost, and improvement of separation efficiency are the main requirements in the development of high-throughput assay methods in bioanalysis. It can be achieved either by ultra-high-performance liquid chromatography (UHPLC) using stationary phases with small particles (<2 μm) at high back pressures or by using opposite direction--monolithic stationary phases with low back pressures. The application of new types of monolithic stationary phases for UHPLC is a novel idea combining these two different paths. The aim of this work was to test the recently introduced second-generation of monolithic column Chromolith® HighResolution for UHPLC analysis of liposoluble vitamins in comparison with core-shell and fully porous sub-2 μm columns with different particle sizes, column lengths, and shapes. The separation efficiency, peak shape, resolution, time of analysis, consumption of mobile phase, and lifetime of columns were calculated and compared. The main purpose of the study was to find a new, not only economical option of separation of liposoluble vitamins for routine practice.

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Miniaturisation of solid phase extraction method for determination of retinol, alpha- and gamma-tocopherol in human serum using new technologies

Cited by 11 publications

References 21 publications

Megaporous monolithic adsorbents for bioproduct recovery as prepared on the basis of nonwoven fabrics

Megaporous monolithic adsorbents for bioproduct recovery as prepared on the basis of nonwoven fabrics

Rapid determination of lipophilic vitamins in human serum by ultra‐high performance liquid chromatography using a fluorinated column and high‐throughput miniaturized liquid–liquid extraction

Comparison of a new high-resolution monolithic column with core-shell and fully porous columns for the analysis of retinol and α-tocopherol in human serum and breast milk by ultra-high-performance liquid chromatography†

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