Analysis and Determination of Ceramide in Konjac by HPLC-ELSD

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.