Flow field-flow fractionation/multiangle light scattering of sodium hyaluronate from various degradation processes

Moon, Myeong Hee; Shin, Dong‐Hee; Lee, Namkoo; Hwang, Euijin; Cho, Il Hwan

doi:10.1016/j.jchromb.2008.01.023

Cited by 15 publications

(12 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…For instance, the same control sample used in Fig. 7 is treated with testicular hyaluronidase by varying the enzyme unit per mL of solution: 2 and 5 U/mL [36]. For each enzymatic reaction, the reaction time was selected to be either 0.5 or 1 h, resulting in four degraded samples labeled 2 U-0.5 h, 2 U-1 h, 5 U-0.5 h, and 5 U-1 h. Fractograms of the four degraded samples are plotted in Fig.…”

Section: Effect Of Enzymatic Reactionmentioning

confidence: 99%

“…FlFFF separation of the control sample was carried out using field decay program-I, and the two degraded samples were run using decay program-II, both of which are provided at the top of the figure. Permission from [36]. slope values are 0.61 and 0.63 for the 2 U samples, denoting unfolding of entangled NaHA molecules.…”

Section: Effect Of Enzymatic Reactionmentioning

confidence: 99%

“…(A) Influence of enzymatic degradation on MWDs of NaHA samples corresponding to the fractograms shown in Figure 9, and (B) plots of RMS radius versus MW values for the enzymatically degraded NaHA products. Permission from [36]. Figure 12A shows the FlFFF fractograms of F3 and F4 before the thermal treatment (at the top) and of F3/T and F4/T after thermal treatment.…”

Section: Effect Of Thermal Treatmentmentioning

confidence: 99%

See 2 more Smart Citations

Flow field‐flow fractionation and multiangle light scattering for ultrahigh molecular weight sodium hyaluronate characterization

Moon

2010

J of Separation Science

Self Cite

View full text Add to dashboard Cite

This review describes the utility of flow field-flow fractionation coupled with multiangle light scattering and differential refractive index (FlFFF-MALS-DRI) detection methods for the separation of ultrahigh molecular weight sodium hyaluronate (NaHA) materials and for the characterization of molecular weight distribution as well as structural determination. The sodium salt of hyaluronic acid (HA), NaHA, is a water-soluble polysaccharide with a broad range of molecular weights (10(5) -10(8) ) found in various naturally occurring fluids and tissues. Basic principles of FlFFF-MALS using field programming for the separation of the degraded products of NaHA prepared by treating raw materials with depolymerization or degradation processes such as membrane filtration, enzymatic degradation, ultrasonic degradation, alkaline reaction, irradiation by γ-rays, and thermal treatment for the development of pharmaceutical applications are introduced. Changes in molecular weight distribution and conformation of NaHA materials due to external stimuli are also discussed.

show abstract

Section: Effect Of Enzymatic Reactionmentioning

confidence: 99%

Section: Effect Of Enzymatic Reactionmentioning

confidence: 99%

Section: Effect Of Thermal Treatmentmentioning

confidence: 99%

See 1 more Smart Citation

Flow field‐flow fractionation and multiangle light scattering for ultrahigh molecular weight sodium hyaluronate characterization

Moon

2010

J of Separation Science

Self Cite

View full text Add to dashboard Cite

show abstract

“…It has been shown that, in practice, FlFFF gives better resolution for MW higher than about 10 5 43. FlFFF has shown its applicability for separation of HA or HA‐like molecules 44–50, for which the frit‐inlet FlFFF, another variant of FlFFF, was mostly 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

View full text Add to dashboard Cite

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.

show abstract

“…The onlinecoupling of the multi angle light scattering (MALS) will also improve AsFlFFF by allowing absolute determination of the molecular weight, sizes, and the molecular conformations of the proteins. [46][47][48][49] With further improvements, and fine-tuning, AsFlFFF is expected to have wide application in the routine analysis of whey proteins. …”

Section: Discussionmentioning

confidence: 99%

Comparison of Size-Exclusion Chromatography and Flow Field-Flow Fractionation for Separation of Whey Proteins

Kang¹,

Moon²,

Lee³

2011

Bulletin of the Korean Chemical Society

View full text Add to dashboard Cite

Whey protein (WP) is a mixture of proteins, and is of high nutritional values. WP has become an important source of functional ingredients in various health-promoting foods. In this study, size-exclusion chromatography (SEC) and asymmetrical flow field-flow fractionation (AsFlFFF) were used for separation and analysis of whey proteins. It was found that a lab-prepared WP from raw milk is mostly of β-lactoglobulin with small amount of higher molecular weight components, while a commercial whey protein isolate (WPI) powder contains relatively larger amount of components other than β-lactoglobulin, including IgG and protein aggregates. Results suggest that AsFlFFF provides higher resolution for the major whey proteins than SEC in their normal operation conditions. AsFlFFF could differentiate the BSA and Albumin, despite a small difference in their molecular weights, and also was able to separate much smaller amount of aggregates from monomers. It is noted that SEC was able to show the presence of low molecular weight components other than the major whey proteins in the WP samples, which AsFlFFF could not show, probably due to the partial loss of those low molecular weight species through the membrane.

show abstract

Flow field-flow fractionation/multiangle light scattering of sodium hyaluronate from various degradation processes

Cited by 15 publications

References 29 publications

Flow field‐flow fractionation and multiangle light scattering for ultrahigh molecular weight sodium hyaluronate characterization

Flow field‐flow fractionation and multiangle light scattering for ultrahigh molecular weight sodium hyaluronate characterization

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

Comparison of Size-Exclusion Chromatography and Flow Field-Flow Fractionation for Separation of Whey Proteins

Contact Info

Product

Resources

About