高效液相色谱-蒸发光散射法测定脂质体中神经酰胺NP和AP含量  

Determination of ceramide NP and AP content in liposomes by high performance liquid chromatography-evaporative light scattering

作　　者：赵格格 苏温柔 王飞飞 Gege Zhao;Wenrou Su;Feifei Wang(Yunnan Botanee Bio-technology Group Co.,Ltd.,Kunming,Yunnan 650106,China;Yunnan Yunke Characteristic Plant Extraction Laboratory Co.,Ltd.,Kunming,Yunnan 650106,China;Medaesthee(Shanghai)Biotechnology Co.,Ltd.,Shanghai 201700,China)

机构地区：[1]云南贝泰妮生物科技集团股份有限公司,云南昆明650106 [2]云南云科特色植物提取实验室有限公司,云南昆明650106 [3]美达丝(上海)生物科技有限公司,上海201700

出　　处：《日用化学工业（中英文）》2023年第12期1466-1471,共6页China Surfactant Detergent & Cosmetics

基　　金：云南特色植物提取实验室自主研究项目基金资助(2022YKZY003)。

摘　　要：建立了高效液相色谱-蒸发光散射法检测神经酰胺脂质体中神经酰胺NP和神经酰胺AP两种成分含量的方法。神经酰胺脂质体原料样品经甲醇-异丙醇(体积比7∶3)破乳并超声处理,采用色谱柱Agilent ZORBAX StableBond C8(4.6×150 mm,3.5μm)分离,以纯乙腈为流动相A,含0.1%甲酸的甲醇-异丙醇(体积比7∶3)为流动相B进行梯度洗脱,流速为1.0 mL/min,柱温40℃,蒸发器温度90℃,雾化器温度70℃,氮气流速1.0 L/min。结果表明,神经酰胺脂质体中NP和AP的定量限分别为3和6μg/mL,检出限分别为1和2μg/mL,重复性和精密度RSD均小于2%,在NP为50~500μg/mL和AP为25~250μg/mL的质量浓度范围内线性关系良好(R^(2)>0.999),不同质量浓度水平下的回收率为96.2%~105.1%。该方法简便可靠,重现性好,可用于不同脂质体中神经酰胺NP和AP的分离与含量测定。Simultaneous chromatographic separations of ceramide NP and ceramide AP in ceramide liposomes were carried out by high-performance liquid chromatography(HPLC)coupled to evaporative light scattering detector(ELSD).The ceramide liposomes samples were treated with methanol-isopropanol(volume ratio 7∶3)and analyzed by Agilent ZORBAX StableBond C8(4.6×150 mm,3.5μm)column.The simultaneous separation of ceramide NP and AP was achieved by gradient elution with 0.1%formic acid in methanol-isopropanol(volume ratio 7∶3)and acetonitrile as the mobile phases.The flow rate was 1.0 mL/min and the column temperature was 40℃.Evaporator and nebulizer temperature were set at 90℃and 70℃,and the gas flow rate was 1.0 L/min.The result shows the detection limits of ceramide NP and AP in ceramide liposomes are 3 and 6μg/mL,respectively,and the quantitative limits are 1 and 2μg/mL,respectively.The RSD of test on ceramide NP and AP repeatability and precision are all less than 2%,and the linear relationship of calibration curves for ceramide NP and AP is good(R^(2)>0.999)in the mass concentration range of 50-500μg/mL and 25-250μg/mL,respectively.Moreover,the recovery rates range from 96.2%to 105.1%at different concentration levels.A simple,reproducible and reliable method is provided for the determination of ceramide NP and AP contents in different ceramide liposomes.

关 键 词：神经酰胺NP 神经酰胺AP 脂质体 高效液相色谱法 蒸发光散射检测器 

分 类 号：TQ658[化学工程—精细化工]