Determination of molecular mass values of chondroitin sulfates by fluorophore-assisted carbohydrate electrophoresis (FACE).
Fluorophore-assisted carbohydrate electrophoresis (FACE) was applied to determine the molecular mass (M) values of various chondroitin sulfate (CS) samples. After labeling with 8-aminonaphthalene-1,3,6-trisulfonic acid (ANTS), FACE was able to resolve each CS sample as a discrete band depending on the M value. After densitometric acquisition, the migration distance of each CS standard was acquired and the third grade polynomial calibration standard curve was determined by plotting the logarithms of the M values as a function of migration ratio. Purified CS samples of different origin and the European Pharmacopeia CS standard were analyzed by both FACE and conventional high-performance size-exclusion liquid chromatography (HPSEC) methods. The molecular weight value on the top of the chromatographic peak (M(p)), the number-average M(n), weight-average M(w), and polydispersity (M(w)/M(n)) were examined by both techniques and found to be quite similar. This study demonstrates that FACE analysis is a suitable, sensitive and simple method for the determination of the M values of CS macromolecules with possible utilization in virtually any kind of research and development such as quality control laboratories.
Buzzega
Dania
D
Department of Biologia Animale, University of Modena and Reggio Emilia, Via Campi 213/d, 41100 Modena, Italy.
Maccari
Francesca
F
Volpi
Nicola
N
D000818:Animals; D002138:Calibration; D002417:Cattle; D002645:Chickens; D002809:Chondroitin Sulfates / Q000737:chemistry*; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004586:Electrophoresis / Q000592:standards; D005456:Fluorescent Dyes / Q000737:chemistry*; D008970:Molecular Weight; D009281:Naphthalenes / Q000737:chemistry*; D012015:Reference Standards; D015203:Reproducibility of Results; D013552:Swine
D005456:Fluorescent Dyes; D009281:Naphthalenes; D002809:Chondroitin Sulfates; C046465:8-amino-1,3,6-naphthalenetrisulfonic acid