氨基葡萄糖3种含量测定方法的比较

The Comparative of Three Determination Methods of Glucosamine

关键词:  氨基葡萄糖  含量测定  USP法  HPLC衍生化法  UV法

Key words:  glucosamine  determination  USP method  HPLC derivatization method  UV method

摘要:

目的比较了USP法、HPLC衍生化法和紫外分光光度法测定氨基葡萄糖含量的准确性和可靠性。方法①USP法参照USP28，采用C8色谱柱（250 mm×4.6 mm, 5 μm），以乙腈-0.05%磷酸溶液（调pH 3.0）（40∶60）为流动相，检测波长195 nm，柱温：30 ℃；②HPLC衍生化法采用C18色谱柱(250 mm×4.6 mm，5 μm)，以乙腈-0.05%磷酸溶液（调pH 3.0）为流动相进行梯度洗脱，检测波长265 nm，柱温：30 ℃，氨基葡萄糖与芴甲氧羰酰琥珀酰亚胺（FMOC-Osu）溶液反应的衍生物在上述条件下检测；③紫外分光光度法采用在碳酸钠溶液环境下铁氰化钾与氨基葡萄糖反应后在420 nm处测定吸收值。结果3种方法的线性均较好， HPLC衍生化法和UV法测定原料药的精密度和回收率均比USP方法好。USP法测定氨基葡萄糖硫酸盐及其制剂的含量偏低。结论USP方法测定结果不理想，HPLC衍生化法和UV法均适合氨基葡萄糖原料的测定，而HPLC衍生化法更适合制剂的含量测定。

Abstract:

OBJECTIVE To compare determination of glucosamine using USP method, HPLC derivitization method and UV method, and inspect their actuality and reliability. METHODS ①Refer to USP28,C8 column(250 mm×4.6 mm, 5 μm) was used with acetonitrile-phosphate buffer(pH 3.0)(40∶60) as mobile phase, detect wavelength was 195 nm and column temperature was 30 ℃. ②HPLC derivitization method used C18 column(250 mm×4.6 mm, 5 μm), the mobile phase consisted of acetonitrile-phosphate buffer(pH 2.5)(40∶60), the detected wavelength of 265 nm, and the temperature of 30 ℃. Glucosamine was derivatized with 9-fluorenylmethoxycarbonyl succinimide (FMOC-Osu) solution and the derivative was detected under the chromatography conditions. ③In the UV method, potassium ferricyanide reacted with glucosamine in sodium carbonate solution and determine the absorption value at 420 nm. RESULTS The linearity of three methods was good. The precision and recovery of HPLC derivitization method and UV method were better than USP method. The result of USP determination of glucosamine sulfate and its preparation was lower. CONCLUSION The result of USP method is not perfect. HPLC derivitization method and UV method are applied to determine the content of drug substances of glucosamine, and the first is more suitable to determination of glucosamine preparation.