Evaluation of in-vitro degradation rate of hyaluronic acid-based hydrogel cross-linked with 1, 4-butanediol diglycidyl ether (BDDE) using RP-HPLC and UV–Vis spectroscopy

A hyaluronic acid (HA) was cross-linked with 1, 4-butanediol diglycidyl ether to produce nine BDDE-HA hydrogels. The degradation rates of six hydrogels were evaluated by HPLC and UV–Visible spectroscopy. The percentage amount of N-acetyl glucosamine NAG obtained after one-day enzymatic digestion to the total amount obtained after complete digestion was an indicative of the degradation rate of each hydrogel. The results were calculated with 95% confidence interval and showed 62.6% ± 12.3 w/w, precision value % R.S.D = 7.95, average recovery = 81.0%, LOD = 6.4 μg/ml for HPLC and 63.3 ± 13.9 w/w, precision value % R.S.D = 8.83, average recovery = 83.1%, LOD = 5.4 μg/ml for UV method. The two methods showed also good linearity with correlation coefficients R2 of 0.998 and 0.9995 for HPLC and UV method respectively. For a comparison purpose, the other three hydrogels were rated using the conventional weight loss method which showed relatively higher degradation rates with an average of 73.4% ± 5.7 w/w, % R.S.D = 3.13. Statistical analysis revealed that there was no significant difference between HPLC and UV–Visible methods, however, these values differed significantly (p < 0.05) from the value obtained from the weight loss method.

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