Turbidimetrische Methode (TBSA) zur Gehaltsbestimmung von Hyaluronsäure
Originale
KEY WORDS Hyaluronsäure | Gehaltsbestimmung | Turbidimetrische Methode mit BSA | Analytische Methode
Abstract
Conventional methods, such as HPLC or the carbazole method are used to determine the content of hyaluronic acid indirectly through their degradation products (monomers and oligomers). A new, simple and rapid turbidimetric method using bovine serum albumin for the determination of hyaluronic acid in pharmaceutical and medical products, like liquid and gel-like formulations based on Dorfman [15] was developed. The objective of this work was to test the factors influencing the turbidimetric method and determine the content of native hyaluronic acid in solid, semi-solid and liquid formulations. The method revealed to be linear from 0.1 mg/ml to 0.7 mg/ml (R ≥ 0.995). The incubation time with bovine serum albumin was set to 10 min. While the use of hyaluronic acid with different molecular weights showed only a little effect on the turbidimetric method, the method highly depends on pH: too acid or basic pH values of the hyaluronic acid samples had a negative impact on the reaction. The pH value of 5.3 revealed to be optimal for the turbidimetric method. The turbidimetric method was robust against the influence of slight variations like various reagents (different batches and manufacturers), different BSA concentrations, temperatures and standing time of analytical solutions. So the turbidimetric method represents an alternative to conventional methods for the quantification of hyaluronic acid in liquid and gel-like products.
Korrespondenz:
M. Sc. Barbara Weber, Labor L+S AG, Mangelsfeld 4,5,6, 97708 Bad Bocklet-Großenbrach, Germany; e-mail: barbara.weber@labor-ls.de
Zusammenfassung
Mit gängigen Methoden wie HPLC oder der Carbazol-Methode lässt sich der Gehalt von Hyaluronsäure (HA) nur indirekt über deren Abbauprodukte (Monomere und Oligomere) bestimmen. Deshalb wurde eine neue, einfache und schnelle turbidimetrische Methode zur Gehaltsbestimmung von nativer Hyaluronsäure in pharmazeutischen und medizinischen Produkten in Anlehnung an Dorfman [15] entwickelt. Ziel der Arbeit war die Austestung von diversen Einflussfaktoren und Grenzen
