HPLC-ELSD测定肠外营养用海豹油脂肪乳中溶血磷脂酰胆碱的含量

Content Determination of Lysophosphatidylcholine in Seal Oil Based Lipid Emulsion for Parenteral Nutrition by HPLC-ELSD

目的建立HPLC-ELSD测定肠外营养用海豹油脂肪乳中溶血磷脂酰胆碱的含量。方法采用硅胶色谱柱（MZ-ANALYTICAL Spherisorb,250 mm×4.6 mm,5μm）,流动相A为甲醇-水-冰醋酸-三乙胺（850∶1 50∶4.5∶0.5）,流动相B为甲醇-水-冰醋酸-三乙胺-正己烷-异丙醇（385∶68∶2∶0.23∶1 60∶3 84）,等度洗脱,A∶B=30∶70,流速为1.0mL.min1;进样体积为20μL,柱温为40℃;蒸发光散射检测器,漂移管温度为70℃,雾化气为氮气,气体流量为2 L.min1。结果线性范围为0.02~0.4 mg.mL1（r=0.999 4）,检测限为0.12μg（S/N=3）,定量限为0.40μg（S/N=10,n=3）,平均回收率为100.3%,RSD为0.54%（n=9）。结论该方法灵敏、快速、准确、重复性好、专属性强,可用于本制剂的质量控制。

OBJECTIVE To establish an HPLC method for determining the content of lysophosphatidylcholine in seal oil based lipid emulsion for parenteral nutrition.METHODS Using a silicone column（MZ-ANALYTICAL Spherisorb,250 mm×4.6 mm,5 μm） as the stationary phase,the mobile phase A was： methanol-water-glacial acetic acid-triethylamine（850∶150∶4.5∶0.5）,and the mobile phase B was： methanol-water-glacial acetic acid-triethylamine-n-hexane-isopropanol（385∶68∶2∶0.23∶160∶384）,and the ratio of A and B was 30∶70 with isocratic elution.The flow rate was 1.0 mL·min 1.The injection volume was 20 μL.The column temperature was set at 40 ℃.The detector was evaporative light-scattering detector（ELSD）.The drift tube temperature of the ELSD was set at 70 ℃,and the nitrogen flow rate of the ELSD was 2 L·min 1.RESULTS The linear concentration range of lysophosphatidylcholine was 0.02 0.4 mg·mL 1（r=0.999 4）,and the average recovery was 100.3% with the RSD of 0.54%（n=9）.The limit of detection was 0.12 μg（S/N=3）,and the limit of quantitation was 0.40 μg（S/N=10,n=3）.CONCLUSION This method is rapid,accurate,reliable and reproducible.It is suitable for the quality control of seal oil based lipid emulsion for parenteral nutrition.