魔芋中神经酰胺类物质的HPLC-ELSD分析及其含量测定  

Analysis and Determination of Ceramide in Konjac by HPLC-ELSD

作　　者：刘仁萍[1] 杨建奎[2] 詹逸舒[1] 曾炽[1] 肖鹜轩 吴永尧[1] 

机构地区：[1]湖南农业大学生物科学技术学院,长沙410128 [2]湖南农业大学理学院,长沙410128 [3]湖南省农业科学研究院,长沙41012

出　　处：《中国生物化学与分子生物学报》2010年第2期189-194,共6页Chinese Journal of Biochemistry and Molecular Biology

基　　金：湖南省科技计划重大专项(No.04NK1002)~~

摘　　要：建立高效液相色谱-蒸发光散射检测器分析神经酰胺的方法并进行了含量的测定.色谱柱:ZORBZX Eclipse XDB-C18(4.6mm×250mm,5μm),洗脱方法:梯度洗脱,柱温:35℃,流动相:甲醇/水,流速:1ml/min;检测器:蒸发光散射检测器,漂移管温度:40℃,氮气流速:1.5L/min.系统探讨了梯度洗脱的起始浓度、洗脱的时间和洗脱梯度的程序设置,最佳的梯度洗脱条件为5min内,甲醇浓度从60%线性增加为90%,从5min到25min,甲醇浓度线性增加为95%,在此条件下样品和标准品的分离色谱峰对称性较好.随后测定了各种样品中神经酰胺的含量,并进行了方法学验证,结果神经酰胺在0.2～2μg之间线性关系良好,最低检测限为0.01mg/ml,R2=0.9992;平均回收率为93.3%,RSD=1.65%(n=5).本法灵敏、方便、准确,重现性好,可用于魔芋神经酰胺类物质的分离及其含量的测定.An analytical method using high-performance liquid chromatography（HPLC） with evaporative light scattering detection（ELSD） has been developed for the determination of ceramide extracted from konjac.The simultaneous separation of ceramide was achieved on a ZORBZX Eclipse XDB-C18 column（4.6 mm×250 mm,5 μm） by gradient elution with methanol-water as the mobile phase at the flow rate of 1 ml/min and the column temperature at 35 ℃.The parameters for ELSD were set by the drift tube temperature 40 ℃ and nitrogen flow-rate 1.5 L/min.After a serial of trials,the optimum elution process was by methanol concentration starting from 60% to 90% in 5min,and then to 95% in 20 min.Under the optimized conditions,the logarithmic linear curve can be obtained from 0.2 to 2 μg with R2 of 0.999 2 and the detection limit（S/N=3） could reach 0.01 mg/ml.The average recovery was 93.3% with RSD =1.65%（n=5）.The results demonstrate that this method provides good reproducibility and sensitivity for the quantification and it is simple,reliable and suitable for the quality control of ceramide extracted from konjac.

关 键 词：神经酰胺 高效液相色谱法 蒸发光散射检测器 

分 类 号：R917.720.1[医药卫生—药物分析学]