离子对HPLC法测定N-乙酰-D-葡萄糖胺的含量

Ion pair HPLC determination of N-acetyl-D-glucosamine

目的:建立离子对 HPLC 法测定 N-乙酰-D-葡萄糖胺及其制剂含量的方法。方法:采用 Alltima ODS 色谱柱(5μm,250 mm×4.6 mm);流动相为含10 mmol·L^(-1)四丁基硫酸氢铵的磷酸氢二钾(0.1 mol·L^(-1))溶液,用磷酸调 pH 至6.5±0.1;流速0.8 mL·min^(-1);柱温:室温;波长:220 nm。结果:在0.4～8μg的范围内 N-乙酰-D-葡萄糖胺峰面积与进样量呈良好的线性关系,r=0.9999;最低检出限为1.3 ng;最低定量限为4 ng;高、中、低3种浓度的回收率分别为99.9%,100.1%,100.2%;RSD 分别为0.46%,0.32%,0.28%。结论:该方法简单、灵敏、准确、专属性好。

Objective ： To establish an HPLC method for the determination of N - Acetyl - D - Glucosamine （ Glc- NAc ）. Methods：Alltima ODS coloum （5 μm, 250 mm × 4.6 mm） was adopted with 0. 1 mmol·L^-1 potassium phosphate dibasic solution （ containing 10 mmol·L^-1 tetrabutylammonium hydrogen sulfate, adjusted pH to 6.5 ± 0. 1 with phosphoric acid ） as the mobile phase; the flow rate was 0. 8 mL ·min^-1 ; coloum temperature was room temperture; the detection wavelength was at 220 nm;Results：A satisfactory separation among GluNAc, related impurities and excipient was obtained. The calibration curve was linear in the range of 0. 4 - 8 μg for GlcNAc. The average recoveries of low, middle and high three concentration from sample were 99.9% , 100. 1% , 100.2% with RSD 2.7%, 0. 03%, 0. 17%,respectively. Conclusion：The method is simple, rapid, accurate and is suitable for the determination of GlcNAc.