Determination of hyaluronic acid by high-performance liquid chromatography of the oligosaccharides derived therefrom as 1-(4-methoxy)phenyl-3-methyl-5-pyrazolone derivatives
1993; Elsevier BV; Volume: 630; Issue: 1-2 Linguagem: Inglês
10.1016/0021-9673(93)80449-i
ISSN1873-3778
AutoresKazuaki Kakehi, Minako Ueda, Shigeo Suzuki, Susumu Honda,
Tópico(s)Polysaccharides and Plant Cell Walls
ResumoHyaluronic acid (HA) was digested with various kinds of depolymerizing enzymes and the products were analysed by high-performance liquid chromatography (HPLC) after derivatization with 1-(4-methoxy)phenyl-3-methyl-5-pyrazolone (PMPMP). As hyaluronate 4-glycanohydrolase (EC 3.2.1.35) from sheep testis showed a high efficiency for depolymerization, giving the tetra- and hexasaccharides abundantly, and is inexpensive, a method for the specific determination of HA was established, based on digestion by this enzyme followed by determination of the tetra- or hexasaccharide derived therefrom as the PMPMP derivatives by HPLC with UV detection. This method allowed the determination of HA in the range 0.5–50 μg with high reproducibility.
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