Determination of glucosamine and its derivatives released from photocrosslinked gelatin hydrogels using HPLC

Abstract

A simple, accurate and validated reverse-phase high-performance liquid chromatography (HPLC)/UV method is developed for the determination of glucosamine hydrochloride (GlcN), N-acetyl-glucosamine (NAG) and N-acryloyl-glucosamine (AGA) released from photocrosslinked gelatin hydrogels. The HPLC separation was achieved on a Shimadzu InertSustain amino column (250 × 4.6 mm, 5 µm particle size) at room temperature using a mobile phase of acetonitrile-phosphate buffer (75:25, v/v, pH 6.0) at a flow rate of 1.0 mL/min and UV detection of 194 nm. The method was validated for specificity, linearity, limit of detection and quantification, accuracy, precision, extraction recovery and solution stability. The calibration curves were with excellent linearity, with correlation coefficients (R(2)) >0.999 for all three drugs. The intra- and inter-day variation was <3.10% and the relative error was between -1.43 and 1.78%. The extraction recovery results ranged from 94.62 to 99.33%, demonstrating the absence of matrix effect. The sample and standard solutions were stable for more than 2 months. The method was successfully used for the analysis of released properties of drugs physically encapsulated and chemically crosslinked in the gelatin hydrogels.

Dasatinib Promotes Chondrogenic Differentiation of Human Mesenchymal Stem Cells via the Src/Hippo-YAP Signaling Pathway

“…The hydrogel with 10% GelMA had a mass swelling ratio of 16.4 ± 1.7 and a compressive modulus of 32.4 ± 3.8 kPa (Figure S3). Based on our previous research, , we used GelMA hydrogel as a carrier for sustained drug release. To measure the release rate of different concentrations of dasatinib from GelMA hydrogels in vitro over 28 days at 37 °C, the HPLC system was used.…”

“…Methacrylate gelatin (GelMA) was synthesized as described in the literature . In brief, 10% (w/v) gelatin foam was dissolved in PBS at 60 °C.…”

Glucosamine‐grafted methacrylated gelatin hydrogels as potential biomaterials for cartilage repair

“…The percentage of grafted AGA in GelMA hydrogels through photocrosslinking was presented by the amount of released AGA determined using the high‐performance liquid chromatography (HPLC) (Suo, Xu, Zhang, & Zheng, ). The GelMA–AGA and GelMA/GlcN hydrogel disks were placed in measuring cups with 1 mL water in a shaking bath.…”

“…At each time point, the supernatant was collected for evaluation. The concentrations of GlcN and AGA were determined by HPLC based on the regression equation as described before (Suo et al, ). Thus, the percentage of grafted AGA in GelMA hydrogels is obtained by ( W i – W r )/ W r , where W i is the initial weight of added drug and W r is the weight of released drug.…”

Determination of glucosamine and monitoring of its mutarotation by hydrophilic interaction liquid chromatography with evaporative light scattering detector

“…HILIC for glucosamine determination has also been reported on a propyl-amino column with refractive index detection (without derivatization; El-Saharty & Bary, 2002;Xinmin, Ruili, Zhihua, Yuanhong, & Tingfu, 2008), or with UV-vis detection after derivatization (Liang, Leslie, Adebowale, Ashraf, & Eddington, 1999). Some researchers have used the propyl-amino column under HILIC mode to detect and quantify glucosamine with direct UV detection at short wavelengths (Shao, Alluri, Mummert, Koetter, & Lech, 2004;Suo, Xu, Zhang, & Zheng, 2016), recently also with MS detection (Roda et al, 2006;Pastorini et al, 2009;Pastorini et al, 2011).…”