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

关键词:  高效液相色谱法  蒸发光散射检测器  海豹油  脂肪乳  溶血磷脂酰胆碱  含量测定

Key words:  HPLC  ELSD  seal oil  lipid emulsion  lysophosphatidylcholine  content determination

摘要:

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，5 μm)，流动相A为甲醇-水-冰醋酸-三乙胺(850∶150∶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∶160∶384)，等度洗脱，A∶B = 30∶70，流速为1.0 mL·min^-1；进样体积为20 μL，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·min^-1。结果 线性范围为0.02~0.4 mg·mL^-1(r=0.999 4)，检测限为0.12 μg(S/N=3)，定量限为0.40 μg(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。

Abstract:

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.