高效阴离子交换色谱法测定那屈肝素钙中2,5-脱水甘露醇和葡萄糖胺

目的： 建立高效阴离子交换色谱法测定那屈肝素钙中2，5-脱水甘露醇和葡萄糖胺。 方法： 样品在3 mol·L^-1 三氟乙酸110℃下水解8 h，旋蒸除酸后用Carbopac PA20 （150 mm×3 mm） 阴离子交换柱在流速0.4 mL·min^-1 和30℃下，以15 mmol·L^-1 氢氧化钠洗脱10 min后用15 mmol·L^-1 氢氧化钠-150 mmol·L^-1 醋酸钠溶液洗脱10 min，脉冲安培检测器检测。 结果： 2，5-脱水甘露醇和葡萄糖胺线性范围分别为1～50 μg·mL^-1 （r=0.9997） 和4.2～124.6 μg·mL^-1 （r=0.9994），检测限分别为163.0 ng·mL^-1 和327.5 ng·mL^-1，回收率（n=6）分别为102.4%和99.5%，RSD均小于3.0%。 结论： 该法简单、灵敏、可靠，可用于那屈肝素钙的质量控制。

Objective: To develop a determination method for 2,5-anhydro mannitol and glucosamine in nadroparin calcium by high performance anion exchange chromatography. Method: The sample was completely hydrolyzed by 3 mol·L^-1 trifluoroacetic acid at 110℃ for 8 hours,deacidified by rotating evaporation,and then analyzed on an analytical column of CarboPac PA20 (150 mm×3 mm) under 30℃ and with a flow rate at 0.4 mL·min^-1.The elution was conducted at 15 mmol·L^-1 sodium hydroxide for 10 min,and then 15 mmol·L^-1 sodium hydroxide-150 mmol·L^-1 sodium acetate for another 10 min.The whole process was monitored by pulsed amperometric detector. Results: The linear ranges of the method for 2,5-anhydro mannitol and glucosamine were 1-50 μg·mL^-1 (r=0.9997) and 4.2-124.6 μg·mL^-1 (r=0.9994),detection limits were 163.0 ng·mL^-1 and 327.5 ng·mL^-1 (based on three times baseline noise),average recoveries (n=6) were 102.4% (RSD=2.2%) and 99.5% (RSD=2.5%),respectively. Conclusion: The established method is simple,sensitive,and reliable,which can be applied to the quality control of nadroparin calcium.