高效液相色谱法测定蜂王浆中6种糖的含量

Determination of the 6 kinds of sugars content in royal jelly by high performance liquid chromatography

目的建立高效液相色谱法测定蜂王浆中6种糖含量的分析方法。方法样品经适量水溶解后,加入乙酸锌、亚铁氰化钾溶液各2.5 mL,涡旋混匀,用水定容至50 mL, 8000 r/min, 4℃离心3 min。滤液过0.45μm液水相膜,采用ZORBAX Carbohydrate Anlys. Col (4.6 mm×150 mm, 5μm)色谱柱,流动相:乙腈-水=70:30 (V:V),等度洗脱,高效液相色谱法测定。结果本方法线性范围分别为葡萄糖1.05~3.0g/100m L、果糖0.84~2.4g/100 mL、蔗糖0.05~0.56 g/100 mL、麦芽糖0.05~0.56 g/100 mL、吡喃葡糖基蔗糖0.005~0.05 g/100 mL、麦芽三糖0.05~0.56 g/100 mL,相关系数分别为果糖1.0000、葡萄糖0.9999、蔗糖0.9997、麦芽糖0.9999、吡喃葡糖基蔗糖0.9990、麦芽三糖0.9998, 6种化合物的检出限为0.2 g/100 g。蜂王浆中糖的回收率为97.0%~101.0%,精密度为2.6%~9.1%。利用该方法测定15个蜂王浆样品,蜂王浆中果糖含量为3.3~7.5g/100g,葡萄糖含量为3.5~8.6 g/100 g,蔗糖含量最高为1.9 g/100 g,麦芽糖含量最高为0.7 g/100 g,吡喃葡糖基蔗糖含量为0.3 g/100 g,麦芽三糖含量小于检出限。结论该法前处理简单、测定快速、准确度高、灵敏度好,可作为标准方法对蜂王浆中葡萄糖、果糖、蔗糖、麦芽糖、吡喃葡糖基蔗糖和麦芽三糖进行快速分析检测。

Objective To establish a method for determination the content of 6 kinds of sugars in royal jelly by high performance liquid chromatography.Methods After the sample was dissolved in proper amount of water,2.5 mL of zinc acetate and 2.5 mL of potassium ferrocyanide solution were added to the sample,and the solution was eddy mixed.The sample was set to 50 mL with constant volume of water,and centrifuged at 8000 r/min for 3 min at 4℃.The filtrate was diluted with 0.45μm liquid/water phase membrane,and the liquid chromatography was performed on a ZORBAX Carbohydrate Anlys.Col(4.6 mm×150 mm,5μm)column with acetonitrile-water=70:30(V:V)as mobile phase.The determination was performed by high performance liquid chromatography.Results The linear ranges of this method were 1.05‒3.0 g/100 mL of fructose,0.84‒2.4 g/100 mL of glucose,0.05‒0.56 g/100 mL of sucrose,0.05‒0.56 g/100 mL of maltose,0.005‒0.05 g/100 mL of erlose,0.05‒0.56 g/100 mL of maltotriose,respectively,and the correlation coefficients were 1.0000,0.9999,0.9997,0.9999,0.9990,0.9998,respectively.The limit of detection the 6 compounds was 0.2 g/100 g.The recoveries of sugars in royal jelly were 97.0%‒101.0%and relative standard deviation was 2.6%‒9.1%.The content of 6 kinds of sugars were 3.3%‒7.5%of fructose,3.5‒8.6 g/100 g of glucose,maximum 1.9 g/100 g of sucrose,maximum 0.7 g/100 g of maltose,maximum 0.3 g/100 g of erlose and maltotriose not detected,respectively in 15 royal jelly samples by this method.Conclusion This method has the advantages of simple pretreatment,rapid determination,high accuracy and good sensitivity.It can be used as a standard method for the rapid analysis and determination of glucose,fructose,sucrose,maltose,pyranosylsucrose and maltotriose in royal jelly.