蒸发光检测器高效液相色谱法测定猪肺磷脂中胆固醇、甘油三油酸酯和甘油三棕榈酸酯  

Determination of CHOL,TG-OL and TG-PAL in porcine pulmonary by high performance liquid chromatography with evaporative luminescence detection

作　　者：郄正刚 曹文倩 王乔波 岳丽娜 耿文飞 马志华[1] 李志永 QIE Zhenggang;CAO Wenqian;WANG Qiaobo;YUE Lina;GENG Wenfei;MA Zhihua;LI Zhiyong(Hebei Changshan Biochemical Pharmaceutical Co.,Ltd.Shijiazhuang 050800,China)

机构地区：[1]河北常山生化药业股份有限公司,河北石家庄050800

出　　处：《化学分析计量》2024年第4期49-53,共5页Chemical Analysis And Meterage

摘　　要：建立蒸发光检测器高效液相色谱测定猪肺磷脂粗品和猪肺磷脂注射液中的胆固醇、甘油三油酸酯和甘油三棕榈酸酯含量。以甲醇-氯仿(体积比为1∶2)为溶剂,采用Caprisil C_(18)-X(250 mm×4.6 mm,5μm)色谱柱,柱温为30℃,进样体积为10μL,流动相A为三氟乙酸体积分数0.83%的乙腈溶液,流动相B为异丙醇,梯度洗脱,流量为1.0 mL/min,蒸发光气体流量为1.8 L/min,漂移管温度为70℃。胆固醇质量浓度的线性范围为10~150μg/mL,甘油三油酸酯和甘油三棕榈酸酯质量浓度的线性范围均为2~60μg/mL。3种目标物的相关系数分别均为0.998,检出限均为1μg/mL,定量限均为2μg/mL。测定结果的相对标准偏差为1.6%~2.3%(n=6),平均加标回收率为108%~113%。该方法可有效控制产品质量,保证已知杂质的有效检出。A high performance liquid chromatography(HPLC)with evaporative luminescence detection(ELSD)method was established for the determination of three related substancessuch as cholesterol(CHOL),glycerol trioleate(TGOL)and glycerol tripalmitate(TG-PAL)in crude extract of porcine pulmonary surfactants and poractant alfa injection.Methanol chloroform(volume ratio 1:2)was used as the solvent,the analytical column was Caprisil C_(18)-X(250 mm×4.6 mm,5μm).The column temperature was 30℃,and the injection volume was 10μL.The mobile phase A was acetonitrile solution with 0.83%(volume fraction)trifluoroacetic acid and the mobile phase B was isopropyl alcohol.Both mobile phases were used for gradient elution,the flow rate was 1.0 mL/min and the gas flow rate of the ELSD was 1.8 L/min with the temperatrure of the drift tube 70℃.The linear range of CHOL mass concentration was 10-150μg/mL,and the linear range of TG-OL and TG-PAL mass concentration was 2-60μg/mL.The correlation coefficients of the three target substances were 0.998,with detection limits of 1μg/mL and quantification limits of 2μg/mL.The relative standard deviations of the determination results were 1.6%-2.3%(n=6),and the average spiked recoveries were 108%~113%.This method can effectively control product quality and ensure the effective detection of known impurities.

关 键 词：猪肺磷脂注射液 胆固醇 甘油三酸油酯 甘油三棕榈酸酯 

分 类 号：O657.7[理学—分析化学]