欢迎访问《日用化学工业（中英文）》，今天是
日用化学工业（中英文）

日用化学工业（中英文） ›› 2023, Vol. 53 ›› Issue (12): 1466-1471.doi: 10.3969/j.issn.2097-2806.2023.12.015

• 分析与检测 • 上一篇    下一篇

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

赵格格1,2,3,苏温柔1,2,3,*,王飞飞1,2,3   

  1. 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
  • 收稿日期:2022-12-05 修回日期:2023-11-29 出版日期:2023-12-22 发布日期:2024-01-12
  • 基金资助:
    云南特色植物提取实验室自主研究项目基金资助(2022YKZY003)

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

Gege Zhao1,2,3,Wenrou Su1,2,3,*,Feifei Wang1,2,3   

  1. 1. Yunnan Botanee Bio-technology Group Co., Ltd., Kunming, Yunnan 650106, China
    2. Yunnan Yunke Characteristic Plant Extraction Laboratory Co., Ltd., Kunming, Yunnan 650106, China
    3. Medaesthee （Shanghai） Biotechnology Co., Ltd., Shanghai 201700, China
  • Received:2022-12-05 Revised:2023-11-29 Online:2023-12-22 Published:2024-01-12
  • Contact: * Tel.: +86-18964202380.

RichHTML

3

PDF (PC)

121

赞

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，3.5 µm）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6 μg/mL，检出限分别为1和2 μg/mL，重复性和精密度RSD均小于2%，在NP为50~500 μg/mL和AP为25~250 μg/mL的质量浓度范围内线性关系良好（R2>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。

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

Abstract:

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, 3.5 μm） 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 6 μg/mL, respectively, and the quantitative limits are 1 and 2 μg/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 （R2>0.999） in the mass concentration range of 50-500 μg/mL and 25-250 μg/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.

Key words: ceramide NP, ceramide AP, liposomes, high-performance liquid chromatography, evaporative light scattering detector

中图分类号: 

  • TQ658