柱前衍生-高效液相色谱法测定保健食品中盐酸氨基葡萄糖含量

Determination of glucosamine hydrochloride content in health foods by pre-column derivatization-high performance liquid chromatography

中文关键词:  盐酸氨基葡萄糖  异硫氰酸苯酯  柱前衍生  液相色谱法  保健食品

英文关键词:glucosamine hydrochloride  phenyl isothiocyanate  pre-column derivatization  high performance liquid chromatography  health food

中文摘要:

      目的 建立以异硫氰酸苯酯为衍生试剂, 采用柱前衍生-高效液相色谱法(high performance liquid chromatography, HPLC)测定保健食品中盐酸氨基葡萄糖含量的方法。方法 样品经水超声提取后, 经异硫氰酸苯酯衍生, 采用Agilent Porshell 120 EC-C18 (4.6 mm×150 mm, 4 μm)色谱柱, 以乙酸钠和甲醇为流动相梯度洗脱, 流速为0.8 mL/min, 在HPLC-紫外检测器245 nm下测定。柱前衍生化条件为: 取1 mL供试品溶液, 加入6 mL磷酸盐缓冲液, 涡旋混匀, 室温放置15 min, 再加入2 mL衍生液置于同一比色管中, 在80 ℃水浴30 min后迅速冷却至室温, 用纯水定容至10 mL。结果 盐酸氨基葡萄糖在25~250 mg/100 g范围内呈良好线性关系(r2=0.9993), 盐酸氨基葡萄糖的检出限为0.0050 mg/mL, 定量限为0.0100 mg/mL, 重复性的相对标准偏差为0.81%, 回收率在97.9%~99.9%。结论 该方法操作简单、结果重复性好, 适用于保健食品中盐酸氨基葡萄糖含量的测定。

英文摘要:

      Objective To establish a method for the determination of glucosamine hydrochloride content in health foods, with phenyl-isothiocyanate as the derivatizing reagent by pre-column derivatization-high performance liquid chromatography (HPLC). Methods Samples were ultrasonically extracted with water, and the extract was derivatized with phenyl-isothiocyanate. Separation was achieved on an Agilent Poroshell 120 EC-C18 column (4.6 mm× 150 mm, 4 μm) via gradient elution with sodium acetate and methanol as mobile phases at a flow rate of 0.8 mL/min. Detection was performed at 245 nm using a HPLC-ultraviolet detector. Pre-column derivatization conditions: 1 mL sample solution was vortex-mixed with 6 mL phosphate buffer, equilibrated at room temperature for 15 min, then mixed with 2 mL phenyl-isothiocyanate reagent in the same colorimetric tube. The mixture was heated in an 80 ℃ water bath for 30 min, rapidly cooled to room temperature, and diluted to 10 mL with water. Results Glucosamine hydrochloride exhibited a good linearity in the range of 25–250 mg/100 g (r2=0.9993). The limit of detection for glucosamine hydrochloride was 0.0050 mg/mL, and the limit of quantitation was 0.0100 mg/mL. The repeatability relative standard deviation was 0.81%, and the recovery rates ranged from 97.9% to 99.9%. Conclusion This method is simple to operate and exhibits good repeatability, making it suitable for the determination of glucosamine hydrochloride content in health foods.