Depolymerization study of sodium hyaluronate by flow field-flow fractionation/multiangle light scattering

被引:13
作者
Kwon, Ji Hye [1 ]
Hwang, Euijin [2 ]
Cho, Il-Hwan [2 ]
Moon, Myeong Hee [1 ]
机构
[1] Yonsei Univ, Dept Chem, Seoul 120749, South Korea
[2] Shinpoong Pharmaceut Co Ltd, Dept Biotechnol, Ansan, Kyeonggi Do, South Korea
关键词
Flow field-flow fractionation; Multiangle light scattering; Sodium hyaluronate; Depolymerization; Biopolymers; Separations/instrumentation; GEL-PERMEATION CHROMATOGRAPHY; FRIT-INLET; MOLECULAR-WEIGHT; PARTICULATE MATERIALS; ACID; ONLINE; DERIVATIVES; CHAINS;
D O I
10.1007/s00216-009-2952-0
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Thermal depolymerization of ultrahigh-molecular-weight (UHMW) sodium hyaluronate (NaHA) was studied systematically by using frit-inlet asymmetrical flow field-flow fractionation/multiangle light scattering/differential refractive index (FI-AFlFFF/MALS/DRI). FI-AFlFFF was utilized for the size separation of NaHA samples which had been thermally degraded for varied treatment times, followed by light-scattering detection to determine MW and structural information of degraded NaHA products. Analysis of NaHA products showed time-dependent depolymerization of raw molecules into smaller-MW components, as well as unfolding of compact structures of UHMW NaHA. To determine whether the observed decrease in MW of sodium hyaluronate originated from the chain degradation of UHMW molecules or from dissociation of entangled complex particles that may have been formed by intermolecular association, narrow size fractions (1 x 10(7)-6 x 10(7) and > 6 x 10(7) MW) of NaHA molecules were collected during FlFFF separation and followed by thermal treatment. Subsequent FI-AFlFFF/MALS analysis of collected fractions after thermal treatment suggested that the ultrahigh-MW region (> 10(7) Da) of NaHA is likely to result from supermolecular structures formed by aggregation of large molecules.
引用
收藏
页码:519 / 525
页数:7
相关论文
共 33 条
[1]   PERSISTENCE LENGTH OF HYALURONIC-ACID - ESTIMATE FROM SMALL-ANGLE X-RAY-SCATTERING AND INTRINSIC-VISCOSITY [J].
CLELAND, RL .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1977, 180 (01) :57-68
[2]   INVITRO STUDIES ON BIOCOMPATIBILITY OF HYALURONIC-ACID ESTERS [J].
CORTIVO, R ;
BRUN, P ;
RASTRELLI, A ;
ABATANGELO, G .
BIOMATERIALS, 1991, 12 (08) :727-730
[3]   Aggregation of amphiphilic pullulan derivatives evidenced by online flow field flow fractionation/multi-angle laser light scattering [J].
Duval, C ;
Le Cerf, D ;
Picton, L ;
Muller, G .
JOURNAL OF CHROMATOGRAPHY B, 2001, 753 (01) :115-122
[5]   FIELD-FLOW FRACTIONATION - ANALYSIS OF MACROMOLECULAR, COLLOIDAL, AND PARTICULATE MATERIALS [J].
GIDDINGS, JC .
SCIENCE, 1993, 260 (5113) :1456-1465
[6]   Flow field-flow fractionation of high-molecular-mass polyacrylamide [J].
Hecker, R ;
Fawell, PD ;
Jefferson, A ;
Farrow, JB .
JOURNAL OF CHROMATOGRAPHY A, 1999, 837 (1-2) :139-151
[7]  
Iqbal Z, 1997, PHARM WORLD SCI, V19, P246, DOI 10.1023/A:1008644400293
[8]  
Kim H, 2006, B KOREAN CHEM SOC, V27, P413
[9]   Field programming in frit inlet asymmetrical flow field-flow fractionation/multiangle light scattering: Application to sodium hyaluronate [J].
Lee, H ;
Kim, H ;
Moon, MH .
JOURNAL OF CHROMATOGRAPHY A, 2005, 1089 (1-2) :203-210
[10]   Effect of dissolution temperature on the structures of sodium hyaluronate by flow field-flow fractionation/multiangle light scattering [J].
Lee, Heejeong ;
Cho, Il-Hwan ;
Moon, Myeong Hee .
JOURNAL OF CHROMATOGRAPHY A, 2006, 1131 (1-2) :185-191