title: Microdetermination of hyaluronan in human plasma by high-performance liquid chromatography with a graphitized carbon column and postcolumn fluorometric detection.
abstract: A chemical method for the determination of hyaluronan (hyaluronic acid, HA) has been developed and applied to the human blood plasma. Human blood plasma HA was converted to the ΔDi-HA by digestion with hyaluronidase SD and determined by a sensitive and selective high-performance liquid chromatography (HPLC). The HPLC includes the separation and detection of ΔDi-HA using a graphitized carbon column and fluorometric reaction with 2-cyanoacetamide in an alkaline eluent. The calibration graph for ΔDi-HA was linear over the range 0.2 ng-1 μg. It was revealed that the concentration of HA in normal human blood plasma is very low levels (about 24 ng/ml) in comparison to low-sulfated chondroitin 4-sulfate (about 13 μg/ml).
authors: Faculty of Pharmaceutical Sciences, Ritsumeikan University, Kusatsu, Shiga, Japan. hidenao@ph.ritsumei.ac.jp
authors: Hidenao
authors: H
authors: Toyoda
authors: Fumie
authors: F
authors: Muraki
authors: Toshio
authors: T
authors: Imanari
authors: Akiko
authors: A
authors: Kinoshita-Toyoda
mesh_terms: D000328:Adult; D002809:Chondroitin Sulfates / Q000737:chemistry / Q000378:metabolism; D002810:Chondroitinases and Chondroitin Lyases / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D005260:Female; D005470:Fluorometry / Q000379:methods*; D006108:Graphite / Q000737:chemistry*; D006801:Humans; D006820:Hyaluronic Acid / Q000097:blood* / Q000737:chemistry / Q000378:metabolism; D006821:Hyaluronoglucosaminidase / Q000378:metabolism; D008297:Male; D009570:Nitriles / Q000737:chemistry / Q000378:metabolism; D012680:Sensitivity and Specificity
chemical_list: D009570:Nitriles; D006108:Graphite; D006820:Hyaluronic Acid; D002809:Chondroitin Sulfates; D006821:Hyaluronoglucosaminidase; D002810:Chondroitinases and Chondroitin Lyases; C031594:2-cyanoacetamide