caa a22 4500 477126723 CHVBK 20180405111652.0 cr unu---uuuuu 170330e19971201xx s 000 0 eng 10.1023/A:1012171511285 doi (NATIONALLICENCE)springer-10.1023/A:1012171511285 Analysis of Drug/Plasma Protein Interactions by Means of Asymmetrical Flow Field-Flow Fractionation [Elektronische Daten] [Maya Madörin, Peter van Hoogevest, Rolf Hilfiker, Birgit Langwost, Gerhard Kresbach, Markus Ehrat, Hans Leuenberger] Purpose. The applicability of Asymmetrical Flow Field-Flow Fractionation (Asymmetrical Flow FFF) as an alternative tool to examine the distribution of a lipophilic drug (N-Benzoyl-staurosporine) within human plasma protein fractions was investigated with respect to high separation speed and loss of material on surfaces due to adsorption. Methods. Field-Flow Fractionation is defined as a group of pseudo-chromatographic separation methods, where compounds are separated under the influence of an externally applied force based on differences in their physicochemical properties. This method was used to separate human plasma in its protein fractions. The drug distribution in the fractions was investigated by monitoring the fractionated eluate for drug content by fluorescence spectroscopy. Results. Human plasma was separated into human serum albumin (HSA), high density lipoprotein (HDL), α2-macroglobulin and low density lipoprotein (LDL) fractions in less than ten minutes. Calibration of the system and identification of the individual fractions was performed using commercially available protein reference standards. The influence of membrane type and carrier solution composition on the absolute recovery of N-Benzoyl-staurosporine and fluorescein-isothio-cyanate-albumin (FITC-albumin) was found to be quite significant. Both factors were optimized during the course of the investigations. N-Benzoyl-staurosporine was found to be enriched in the fraction containing HSA. Conclusions. If experimental conditions are thoroughly selected and controlled to suppress drug and plasma protein adsorption at the separation membrane, Asymmetrical Flow FFF shows high recoveries and fast separation of human plasma proteins, and can be a reliable tool to characterize drug / plasma protein interactions. For analytical purposes it has the potential to rival established technologies like ultracentrifugation in terms of ease-of-use, precision, and separation time. Plenum Publishing Corporation, 1997 asymmetrical flow field-flow fractionation nationallicence FFF nationallicence plasma proteins nationallicence HAS nationallicence lipoproteins nationallicence N-Benzoyl-staurosporine nationallicence recovery nationallicence separation nationallicence Madörin Maya Pharmaceutical and Analytical Development, Novartis Pharma AG, CH-, 4002, Basel, Switzerland aut van Hoogevest Peter Pharmaceutical and Analytical Development, Novartis Pharma AG, CH-, 4002, Basel, Switzerland aut Hilfiker Rolf School of Pharmacy, University of Basel, CH-4051 Basel, Switzerland aut Langwost Birgit BioAnalytical Research, Novartis Pharma AG, CH-4002 Basel, Switzerland aut Kresbach Gerhard BioAnalytical Research, Novartis Pharma AG, CH-4002 Basel, Switzerland aut Ehrat Markus BioAnalytical Research, Novartis Pharma AG, CH-4002 Basel, Switzerland aut Leuenberger Hans School of Pharmacy, University of Basel, CH-o4051, Basel, Switzerland aut Pharmaceutical Research Kluwer Academic Publishers-Plenum Publishers 14/12(1997-12-01), 1706-1712 0724-8741 14:12<1706 1997 14 11095 https://doi.org/10.1023/A:1012171511285 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1012171511285 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Madörin Maya Pharmaceutical and Analytical Development, Novartis Pharma AG, CH-, 4002, Basel, Switzerland aut NATIONALLICENCE 700 1- van Hoogevest Peter Pharmaceutical and Analytical Development, Novartis Pharma AG, CH-, 4002, Basel, Switzerland aut NATIONALLICENCE 700 1- Hilfiker Rolf School of Pharmacy, University of Basel, CH-4051 Basel, Switzerland aut NATIONALLICENCE 700 1- Langwost Birgit BioAnalytical Research, Novartis Pharma AG, CH-4002 Basel, Switzerland aut NATIONALLICENCE 700 1- Kresbach Gerhard BioAnalytical Research, Novartis Pharma AG, CH-4002 Basel, Switzerland aut NATIONALLICENCE 700 1- Ehrat Markus BioAnalytical Research, Novartis Pharma AG, CH-4002 Basel, Switzerland aut NATIONALLICENCE 700 1- Leuenberger Hans School of Pharmacy, University of Basel, CH-o4051, Basel, Switzerland aut NATIONALLICENCE 773 0- Pharmaceutical Research Kluwer Academic Publishers-Plenum Publishers 14/12(1997-12-01), 1706-1712 0724-8741 14:12<1706 1997 14 11095 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer