CAJ | 학술논문

目的：采用高效液相色谱串联质谱法建立测定细胞中不同类型神经酰胺含量的分析方法。方法：采用 Agilent 1290高效液相色谱仪系统， ZORBAX Eclipse XDB－C8（150．0 mm ×2．1 mm，3．5μL）色谱柱，柱温35℃；流动相A为含1 mmol／L甲酸铵和0．2％甲酸的甲醇溶液，流动相 B 为100％甲醇；流速为0．3 mL／min，梯度洗脱；质谱检测中采用电喷雾离子源多反应监测模式。结果：C16、C17、C18、C24四种神经酰胺的色谱响应线性范围良好（ R2≥0．9989）；平均回收率分别为99．1％、99．9％、100．5％、98．2％，相对标准偏差分别为5．6％、5．1％、4．8％、5．5％（ n＝9）。结论：采用高效液相色谱串联质谱法建立同时测定细胞中不同类型神经酰胺的方法灵敏度高，重现性好，可以作为检测细胞中不同类型神经酰胺含量的工具。
목적：채용고효액상색보천련질보법건립측정세포중불동류형신경선알함량적분석방법。방법：채용 Agilent 1290고효액상색보의계통， ZORBAX Eclipse XDB－C8（150．0 mm ×2．1 mm，3．5μL）색보주，주온35℃；류동상A위함1 mmol／L갑산안화0．2％갑산적갑순용액，류동상 B 위100％갑순；류속위0．3 mL／min，제도세탈；질보검측중채용전분무리자원다반응감측모식。결과：C16、C17、C18、C24사충신경선알적색보향응선성범위량호（ R2≥0．9989）；평균회수솔분별위99．1％、99．9％、100．5％、98．2％，상대표준편차분별위5．6％、5．1％、4．8％、5．5％（ n＝9）。결론：채용고효액상색보천련질보법건립동시측정세포중불동류형신경선알적방법령민도고，중현성호，가이작위검측세포중불동류형신경선알함량적공구。
Objective: To establish a high-performance liquid chromatography-tandem mass spectrometry ( HPLC-ESI-MS ) method for simultaneous quantitative analysis of different ceramide species in cells.Methods:The analysis was performed on an Agilent 1290 HPLC system with a ZORBAX Eclipse XDB-C8 (150.0 mm ×2.1 mm, 3.5 μL) column and a temperature of 35 ℃.Methanol with 1 mmol/L ammonium formate and 0.2% formic acid was used as mobile phase A and 100% methanol was used as mobile phase B.And the ceramides were separated by gradient elution at a flow rate of 0.3 mL/min.Electrospray ionization ( ESI ) with multiple reaction monitoring ( MRM) was used in the analysis.Results:Four ceramide species all had a good linear response in the determination ranges (R2≥0.9987).The average recoveries (n=9) were 99.1%,99.9%,100.5% and 98.2% with RSDs of 5.6%, 5.1%, 4.7% and 5.5%, respectively.In addition, the levels of ceramides in FL cells were relatively stable, while the C24-ceramide had the highest level.Conclusion:The HPLC-ESI-MS method for simultaneous analysis of ceramides has high accuracy, reproducibility and linearity, which may be used for quantification of ceramide species in cells.