Mass overloading in the flow field-flow fractionation channel studied by the behaviour of the ultra-large wheat protein glutenin

Arfvidsson, Cecilia; Wahlund, Karl-Gustav

doi:10.1016/s0021-9673(03)01145-2

Cited by 39 publications

(30 citation statements)

References 34 publications

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“…The particles will not reach their correct equilibrium height over the accumulation wall and thus migrate at a larger distance from the accumulation wall and elute earlier than expected from the FFF theory. Similar behavior has previously been reported for other samples, e.g., ultra-large wheat protein glutenin [19], humic substances [20], polystyrene latex particles [25], and for CPMV [21]. For the neutral liposomes studied here, an optimum salt concentration appears to be in the magnitude of 5 -10 mM.…”

Section: Influence Of Ionic Strength Of the Carrier Solutionsupporting

confidence: 87%

“…The retention time is decreased with increase in sample load. Similar observations have been reported for wheat protein glutenin with sodium phosphate buffer as carrier [19], humic substances in distilled water [20], and ferritin and Cowpea mosaic virus (CPMV) in 0.02% NaN 3 Tris-HNO 3 buffer of low ionic strength (0.008 M) [21]. Obviously, with high sample loads some bigger particles elute earlier than expected.…”

Section: Influence Of Sample Loadsupporting

confidence: 80%

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Asymmetric flow field‐flow fractionation of liposomes: optimization of fractionation variables

Hupfeld

Ausbacher

Brandl

2009

J of Separation Science

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Original Paper Asymmetric flow field-flow fractionation of liposomes: optimization of fractionation variablesThe purpose of this study was to investigate the influence of ionic strength of the carrier liquid, cross flow rate, focus flow rate, and sample load on the retention behavior of liposomes in asymmetric flow field-flow fractionation (AF4). Two differently prepared samples of large unilamellar vesicles (LUV) were used. Experiments were performed varying the factors systematically and evaluating their effect on both retention behavior of the liposomes and on particle size as obtained from online coupled multi-angle light scattering (MALS) analysis. The results showed that the focus flow rate had the least influence on the elution of liposomes. Elution of LUV is mainly governed by the chosen cross flow condition and ionic strength of the carrier liquid as well as its sample load. Optimal fractionation and size analysis were achieved using a sample load of about 10 lg, a cross flow gradient from 1.0 to 0.1 mL/min over 35 min and a carrier solution of NaNO 3 with a concentration of 10 mM. IntroductionLiposomes are vesicles consisting of an aqueous core surrounded by a phospholipid bilayer [1]. They have the ability to incorporate both amphiphilic and lipophilic compounds in their membrane-like bilayer as well as to encapsulate hydrophilic compounds [2]. Due to this ability liposomes have contributed to improving drug delivery of e.g., anthracyclines and play an important role in drug delivery research. It is widely accepted that liposomes as drug carriers may improve drug delivery in terms of therapeutic activity and safety [3 -5]. The size distribution of liposomes is important in terms of both the systemic circulation time upon injection into the blood stream and targeting abilities [3,6,7]. A study of the biodistribution of liposomes showed that the average size of the samples does not supply enough information and rather the size distribution should be used, especially for less homogeneous samples [7]. Asymmetric flow FFF (AF4) in combination with multi-angle light scattering (MALS) is the mostly applied sub-class of the FFF family and has gained increasing importance in the field of protein and peptide therapeutics as well as natural and synthetic polymers in recent years [8,9]. However, the number of publications about FFF of liposomes is rather limited compared to other applications and the technique has still not found widespread use despite it has various promising advantages over current particle size analysis methods such as photon correlation spectroscopy (PCS), and size exclusion chromatography [10,11]. For liposomal drug carriers a reliable size determination over the whole particle size spectrum from a few nanometers up to a micrometer could not be achieved with a single method so far. Such information on the other hand is essential for stringent control of the particle size which is governing the biodistribution and tumor targeting [7]. Despite a handful of successful attempts to characterize ...

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Section: Influence Of Ionic Strength Of the Carrier Solutionsupporting

confidence: 87%

Section: Influence Of Sample Loadsupporting

confidence: 80%

Asymmetric flow field‐flow fractionation of liposomes: optimization of fractionation variables

Hupfeld

Ausbacher

Brandl

2009

J of Separation Science

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“…A similar approach with calibration against standard proteins has been reported (Daqiq, Fellows, Bekes, & Lees, 2007). Glutenin, as many ultra-high molar mass macromolecules (see sections on starch, gums and alginates), was sensitive to overloading in the separation channel, calling for a careful optimization of experimental conditions (Arfvidsson & Wahlund, 2003). In a following paper direct molar mass determination (from MALS-RI detection) was achieved and the molar mass range found was 10 4 e10 8 g/mol (Arfvidsson, Wahlund, & Eliasson, 2004).…”

Section: Cereal Proteinsmentioning

confidence: 77%

Separation and characterization of food macromolecules using field-flow fractionation: A review

Nilsson

2013

Food Hydrocolloids

118

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“…In practice, the influence of the particle effective volume was recognized very early. Peak distortion and variation in elution profiles with the increase in the sample load were reported for various samples [23,24,26,36,38]. This effect termed overloading was found to depend on sample characteristics as well as on the composition of the carrier solution.…”

Section: Effect Of Injected Sample Massmentioning

confidence: 94%

Asymmetrical flow field‐flow fractionation coupled to multiangle laser light scattering detector: Optimization of crossflow rate, carrier characteristics, and injected mass in alginate separation

Alasonati¹,

Benincasa²

2007

J of Separation Science

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The coupling of the flow field-flow fractionation (FlFFF) to differential refractive index (DRI) and multiangle laser light scattering (LS) detectors is a powerful tool for characterizing charged polysaccharides such as alginate. However, the correct interpretation of the experimental results and extrapolation of meaningful molecular parameters by using an analytical tool with such a level of complexity requires improvement of the knowledge of the alginate behavior in the channel and careful optimization of the operating conditions. Therefore, the influence of the critical operating parameters, such as crossflow rate, carrier composition and concentration, and sample load, on the alginate retention was carefully evaluated. Combined information obtained simultaneously by DRI and LS detectors over the wide range of the crossflow rate, carrier liquid concentration, and injected amount, allowed to set the appropriate combination of optimal parameters. It was found that the crossflow rate of 0.25 mL/min, carrier solution containing 5x10(-2 )mol/L ammonium or sodium chloride, and 50-100 microg of injected sample mass were necessary to achieve complete separation and determination of the meaningful molecular characteristics. The values of the weight-average hydrodynamic radius (R(Hw)), radius of gyration (R(G)), and molar mass (M), obtained under the optimal conditions were in good agreement to those found for alginates in the literature.

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Mass overloading in the flow field-flow fractionation channel studied by the behaviour of the ultra-large wheat protein glutenin

Cited by 39 publications

References 34 publications

Asymmetric flow field‐flow fractionation of liposomes: optimization of fractionation variables

Asymmetric flow field‐flow fractionation of liposomes: optimization of fractionation variables

Separation and characterization of food macromolecules using field-flow fractionation: A review

Asymmetrical flow field‐flow fractionation coupled to multiangle laser light scattering detector: Optimization of crossflow rate, carrier characteristics, and injected mass in alginate separation

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