高效液相色谱法测定氨基葡萄糖中的乙酰氨基葡萄糖

目的:建立高效液相色谱法测定氨基葡萄糖(GlcN)样品中乙酰氨基葡萄糖(GlcNAc)。方法: 采用氨基柱(250 mm×4.6 mm,5 μm),以乙腈-磷酸盐缓冲液(65:35,pH 7.5)为流动相,流速1.0 mL · min^-1,柱温30 ℃,检测波长195 nm,进样体积20 μL。结果: 方法专属性强,GlcNAc与GlcN 及各杂质分离良好;GlcNAc在1.94~14.70 μg · mL^-1浓度范围内线性良好(r=0.999 9),准确度高(回收率99.0%~104.2%,n=3),溶液在24 h内稳定(RSD为0.69%),精密度(RSD为1.5%~2.2%)和耐用性(RSD为1.6%~8.1%)均良好,灵敏度高(定量限0.98 μg · mL^-1)。经过测定,盐酸氨基葡萄糖粗品中GlcNAc含量为0.09%~0.18%,盐酸氨基葡萄糖成品和硫酸氨基葡萄糖氯化钾成品中未检出GlcNAc(含量<0.0049%)。结论: 本法经方法学验证,能够快速、准确地测定出GlcN中GlcNAc的含量。

Objective: To establish the HPLC method for determination of acetylglucosamine (GlcNAc) in glucosamine (GlcN). Methods: The sample was analyzed on a Hedera NH₂-Pak column(250 mm×4.6 mm, 5 μm)with the mobile phase of acetonitrile-phosphate buffer(65:35, pH 7.5)at the flow rate of 1.0 mL · min^-1 and the column temperature was 30 ℃;the detection wavelength was 195 nm and the injection volume was 20 μL. Results: The analysis method was specific.GlcNAc was separated well from GlcN and its impurities.The linear range of GlcNAc was 1.94-14.70 μg · mL^-1, r=0.999 9;the average recovery(n=3)of GlcNAc was 99.0%-104.2%;the solution was stable in 24 h(RSD=0.69%).Precision(RSD=1.5%-2.2%) and durability(RSD=1.6%-8.1%)of the method were good and the limit of quantitation of GlcNAc was 0.98 μg · mL^-1.The content of GlcNAc in crude glucosamine hydrochloride products was 0.09%-0.18% and GlcNAc(the content<0.0049%)was not detected in the end products of glucosamine hydrochloride and glucosamine sulfate potassium chloride. Conclusion: This verified HPLC method can be applied to quickly and accurately determine GlcNAc in GlcN.