Journal
JOURNAL OF SEPARATION SCIENCE
Volume 33, Issue 22, Pages 3530-3536Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/jssc.201000478
Keywords
Asymmetrical flow field-flow fractionation; Hyaluronic acid; Multiangle light scattering; Molecular weight distribution; Rooster comb
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Funding
- Hannam University
- Ministry of Education, Science and Technology [2010-0003133]
- National Research Foundation of Korea [2010-0003133] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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Hyaluronic acid (HA) was extracted in a relatively large scale from rooster comb using a method similar to that reported previously. The extraction method was modified to simplify and to reduce time and cost in order to accommodate a large-scale extraction. Five hundred grams of frozen rooster combs yielded about 500 mg of dried HA. Extracted HA was characterized using asymmetrical flow field-flow fractionation (AsFlFFF) coupled online to a multiangle light scattering detector and a refractive index detector to determine the molecular size, molecular weight (MW) distribution, and molecular conformation of HA. For characterization of HA, AsFlFFF was operated by a simplified two-step procedure, instead of the conventional three-step procedure, where the first two steps (sample loading and focusing) were combined into one to avoid the adsorption of viscous HA onto the channel membrane. The simplified two-step AsFlFFF yielded reasonably good separations of HA molecules based on their MWs. The weight average MW (M-w) and the average root-mean-square (RMS) radius of HA extracted from rooster comb were 1.20 x 10(6) and 94.7 nm, respectively. When the sample solution was filtered through a 0.45 mu m disposable syringe filter, they were reduced down to 3.8 x 10(5) and 50.1 nm, respectively.
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