Characterization of Fluorescein Isothiocyanate-Dextrans Used in Vesicle Permeability Studies
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- 作者：Karine Andrieux ; Pierre Lesieur ; Sylviane Lesieur ; Michel Ollivon ; and Cé ; cile Grabielle-Madelmont
- 刊名：Analytical Chemistry
- 出版年：2002
- 出版时间：October 15, 2002
- 年：2002
- 卷：74
- 期：20
- 页码：5217 - 5226
- 全文大小：188K
- 年卷期：v.74,no.20(October 15, 2002)
- ISSN：1520-6882
文摘
Fluorescein Isothiocyanate-dextrans of various weightaverage molecular masses (4400-487 000) were analyzed in buffer solution for pH, osmolarity, fluorescenceintensity as a function of the polymer concentration,average molecular masses, and radii of gyration. Labelingof polymers and conformation of the polymers werecharacterized by high-performance gel exclusion chromatography (HPLC-GEC) and small-angle X-ray scattering.The fluorescence measurements evidence the absence offluorescence quenching of the FITC chromophores but theexistence of an inner filter effect at high polymer concentration. The conformation of the polymers in buffer is verylikely of random coil type, as shown by the relationshipbetween the radii of gyration and the weight-averagemolecular masses of the dextrans (
w). The medium usedto analyze the FITC-dextrans by HPLC-GEC stronglyinfluences their elution behavior. In buffer medium, theyare sieved over the TSK G4000 PW column through asingle population according to their w, whereas in purewater, they are separated into several species by anexclusion mechanism that depends on the number oflabeled sites per dextran molecule. A Monte Carlo simulation was used to analyze the distribution of the fluorescentlabels. HPLC-GEC in water could interestingly be appliedto yield labeled polymers bearing a known number offunctionalized groups.
w). The medium usedto analyze the FITC-dextrans by HPLC-GEC stronglyinfluences their elution behavior. In buffer medium, theyare sieved over the TSK G4000 PW column through asingle population according to their w, whereas in purewater, they are separated into several species by anexclusion mechanism that depends on the number oflabeled sites per dextran molecule. A Monte Carlo simulation was used to analyze the distribution of the fluorescentlabels. HPLC-GEC in water could interestingly be appliedto yield labeled polymers bearing a known number offunctionalized groups.