一测多评法同时测定薏苡仁油中4种甘油三酯类抗肿瘤成分的含量

目的:建立同时测定薏苡仁油中4种甘油三酯类抗肿瘤成分含量的方法。方法:采用高效液相色谱-蒸发光散射检测器法(HPLC-ELSD)。色谱柱为Inertsil ODS-3 C₁₈,流动相为乙腈-异丙醇(57∶43,V/V),流速为1.0 mL/min,柱温为30℃,进样量为10μL;蒸发光散射检测器漂移管温度为70℃,气体流速为2 L/min。以甘油三油酸酯为内标,分别计算其与甘油三亚油酸酯、1,2-亚油酸-3-棕榈酸甘油酯、1-棕榈酸-2-油酸-3-亚油酸甘油酯的相对校正因子(RCF),通过RCF计算薏苡仁油中上述3种成分的含量,同时采用外标法测定薏苡仁油中这4种成分的含量,比较一测多评法和外标法的含量测定结果。结果:甘油三亚油酸酯、1,2-亚油酸-3-棕榈酸甘油酯、1-棕榈酸-2-油酸-3-亚油酸甘油酯、甘油三油酸酯进样量检测线性范围分别为0.15～4.50、0.15～4.50、0.35～10.50、0.35～10.50μg(r≥0.999 5);定量限分别为0.13、0.06、0.07、0.12μg,检测限分别为0.04、0.02、0.02、0.03μg;精密度、稳定性、重复性试验的RSD<2.0%(n=6);加样回收率为95.43%～102.67%(RSD<2.0%,n=6)。甘油三亚油酸酯、1,2-亚油酸-3-棕榈酸甘油酯、1-棕榈酸-2-油酸-3-亚油酸甘油酯的平均RCF分别为0.31、0.88、1.21。在不同试验条件下,RCF重现性良好。一测多评法和外标法的含量测定结果比较,差异无统计学意义(P>0.05)。结论:该方法操作简单快捷、结果准确可靠,可用于薏苡仁油中甘油三亚油酸酯、1,2-亚油酸-3-棕榈酸甘油酯、1-棕榈酸-2-油酸-3-亚油酸甘油酯、甘油三油酸酯4种抗肿瘤成分含量的同时测定。

Simultaneous Determination of 4 Triglyceride Anti-tumor Components in Coix lacryma Seed Oil by Quatitative Analysis of Multi-components by Single Marker

OBJECTIVE:To establish a method for simultaneous determination of 4 triglyceride anti-tumor components in Coix lacryma seed oil.METHODS:HPLC-ELSD was adopted.The determination was performed on Inertsil ODS-3 C18 column with mobile phase consisted of acetonitrile-isopropanol(57∶43,V/V)at the flow rate of 1.0 mL/min.The column temperature was 30℃,and the sample size was 10μL.Evaporative light scattering detector was used,the drift tube temperature was 70℃,and the gas flow rate was 2 L/min.Using glycerol trioleateas internal standard,relative correction factors(RCF)of linolein trilinolein,1,2-linoleic acid-3-palmitic acid glyceride and 1-palmitic acid-2-oleic acid-3-linoleic acid glyceride were calculated respectively.The contents of above 3 components in C.lacryma seed oil were calculated by RCF.The contents of 4 components in C.lacryma seed oil were determined by external standard.The results of content determination by quatitative analysis of multi-components by single marker(QAMS)were compared with external standard method.RESULTS:The linear ranges were 0.15-4.50μg for linolein trilinolein,0.15-4.50μg for 1,2-linoleic acid-3-palmitate,0.35-10.50μg for 1-palmitic acid-2-oleic acid-3-linoleic acid glyceride,0.35-10.50μg for glycerol trioleate(r≥0.999 5).The limits of quantification were 0.13,0.06,0.07,0.12μg.The limits of detection were 0.04,0.02,0.02,0.03μg,respectively.RSDs of precision,stability,and repeatability tests were less than 2.0% (n=6).The average recoveries were 95.43% -102.67% (RSD<2.0% ,n=6).Average RCFs of linolein trilinolein,1,2-linoleic acid-3-palmitic acid glyceride and 1-palmitic acid-2-oleic acid-3-linoleic acid glyceride were 0.31,0.88,and 1.21,respectively.RCFs reproducibility was perfect under different experiment conditions.There was no significant difference in results of content determination between QAMS and external standard method(P>0.05).CONCLUSIONS:The method is simple,rapid,accurate and reliable.It is used for simultaneous determination of linolein trilinolein,1,2-linoleic acid-3-palmitate,1-palmitic acid-2-oleic acid-3-linoleic acid glyceride and glycerol trioleateas in C.lacryma seed oil.