高效液相色谱-蒸发光散射检测法测定蛋黄卵磷脂的含量

建立了蛋黄磷脂中卵磷脂（即磷脂酰胆碱,PC)的高效液相色谱-蒸发光散射检测（HPLC-ELSD）的测定方法。以Nov a-Pak Silica 60A硅胶柱(3.9 mm i.d.×150 mm,4 μm)为分离柱,正己烷-异丙醇-3%冰醋酸水溶液(体积比为35∶65 ∶8)为流动相，等度洗脱,流速1.0 mL/min,柱温30 ℃。蒸发光散射检测器漂移管温度50 ℃,雾化气(空气)压力350 kPa。在上述条件下测得PC在0.16~1.61 g/L范围内线性关系良好(r2=0.9979),检测限为0.64 μg,方法的精密度为3.2%(n=5),回收率为98.2%~128.2%。将该方法用于实际样品的测定,获得了令人满意的结果。该方 法预处理简单,分析速度快,可用于蛋黄磷脂中卵磷脂的测定。

Determination of Egg-Yolk Phosphatidylcholine by Normal-Phase High Performance Liquid Chromatography with Evaporative Light Scattering Detection

A method for the determination of egg-yolk phosphatidylcholine by normal-phase high performance liquid chromatography (NP-HPLC) with evaporative light scattering detection (ELSD) was developed. Yolk phosphatidylcholine was successfully separated on a Nova-Pak Silica 60A column (3.9 mm i.d.×150 mm, 4 μm), using hexane-isopropanol-water containing 3% acetic acid (35∶65∶8, v/v/v) as mobile phase at a flow rate of 1.0 mL/min. The drift tube temperature and air carrier gas pressure of the ELSD were set at 50 ℃ and 350 kPa, respectively. The linear detection range was 0.16-1.61 g/L (r2=0.9979) and the limit of detection (LOD) was 0.64 μg with excellent relative standard deviation (RSD) of 3.2% (n=5). The recovery was in the range of 98.2%-128.2%. The results indicated that this method is accurate, rapid, simple and reproducible, and thus suitable for the analysis of egg-yolk phosphatidylcholine in real samples.