魔芋中神经酰胺类物质的HPLC-ELSD分析及其含量测定

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.

Analysis and Determination of Ceramide in Konjac by HPLC-ELSD

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.