High-performance size-exclusion chromatography of proteoglycans extracted from bovine articular cartilage

Dekeyser, Philippe; Smedt, Stefaan De; Vercruysse, Kp; Demeester, Jo; Lauwers, Albert

doi:10.1016/0003-2670(93)85073-s

Cited by 2 publications

(1 citation statement)

References 4 publications

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“…For relatively small HA derivatives or fragments, HPLC 8–12 or HPLC‐MS 13, 14 have been employed. For large underivatized HA, size‐exclusion chromatography (SEC) 15–22, SEC combined with viscometry 23–25, SEC combined with light scattering (LS) detection (SEC‐LS) 23–32, SEC combined with MALDI‐MS (SEC‐MALDI‐MS) 33–35 have been used.…”

Section: Introductionmentioning

confidence: 99%

Extraction of hyaluronic acid (HA) from rooster comb and characterization using flow field‐flow fractionation (FlFFF) coupled with multiangle light scattering (MALS)

Kang

Kim²,

Heo³

et al. 2010

J of Separation Science

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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 1.20×10(6) and 94.7 nm, respectively. When the sample solution was filtered through a 0.45 μm disposable syringe filter, they were reduced down to 3.8×10(5) and 50.1 nm, respectively.

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