Original Article
Development and validation of an LC-MS/MS method for the determination of 12 ceramides in human plasma
Liu Jinsong, Cheng Xiaoliang, He Ziyun, Yan Renqing, Zhang Wei, Yan Shengkai
Published 2023-08-19
Cite as Chin J Lab Med, 2023, 46(8): 830-839. DOI: 10.3760/cma.j.cn114452-20230615-00298
Abstract
ObjectiveTo establish and validate a reliable and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the detection of 12 ceramides in human plasma.
MethodsFrom October 2021 to October 2022, 438 apparently healthy individuals were enrolled in the Affiliated Hospitals of Zunyi Medical University for reference intervals of 12 ceramides in this population. Plasma samples were collected, and separated using the ACQUITY UPLC BEH C18 (2.1×50 mm, 1.7 μm) column, deuterated isotopes were used as internal standards. The mobile phase is water (containing 0.1% formic acid) and isopropanol: acetonitrile (1∶1, v/v, containing 0.1% formic acid) at a flow rate of 0.4 ml/min with gradient elution. The detection method was established using the Qlife Lab 9000 Plus triple quadrupole mass spectrometer. The performance of the method was evaluated in terms of linearity, the lower limit of quantification, precision, recovery, and stability.
ResultsThe method passed the performance evaluation in terms of linearity, the lower limit of quantification, recovery, precision, and stability. The intra-and inter-batch precision of the 12 ceramides ranged from 1.3% to 14.3%, the correctness was verified by spiked recovery experiments, and the recoveries ranged from 91.9% to 111.0%. The lower limit of quantification ranged from 0.001 to 0.100 μmol/L. Standard curve showed good linearity (correlation coefficient r>0.990). Stability tests showed that the 12 ceramides were stable in the biological matrix and after processing under different conditions for a specified period of time. The corresponding biological reference intervals were established for each of the 12 ceramides: 0.103-0.326 μmol/L for Cer(d18∶1/16∶0), 0.018-0.098 μmol/L for Cer(d18∶1/18∶0), 0.933-3.919 μmol/L for Cer(d18∶1/24∶0), 0.243-1.072 μmol/L for Cer(d18∶1/24∶1), 0.001-0.007 μmol/L for Cer(d18∶1/14∶0), 0.022-0.095 μmol/L for Cer(d18∶1/20∶0), 0.185-0.835 μmol/L for Cer(d18∶1/22∶0), 0.003-0.022 μmol/L for Cer(d18∶0/16∶0), 0.001-0.016 μmol/L for Cer(d18∶0/18∶0), 0.017-0.156 μmol/L for Cer(d18∶0/24∶0), 0.008-0.074 μmol/L for Cer(d18∶0/24∶1), and 0.106-0.721 μmol/L for LacCer(d18∶1/24∶1).
ConclusionOur study shows that the newly established LC-MS/MS method for the determination of 12 ceramides in human plasma is reliable, and suitable for clinical application.
Key words:
Ceramide; Chromatography, liquid; Tandem mass spectrometry
Contributor Information
Liu Jinsong
Department of Laboratory Medicine, Affiliated Hospital of Zunyi Medical University, Zunyi 563003, China
College of Laboratory Medicine, Zunyi Medical University, Zunyi 563006, China
Cheng Xiaoliang
Jiangsu Qlife Medical Technology Group Co., LTD., Nanjing 211899, China
He Ziyun
Department of Laboratory Medicine, Affiliated Hospital of Zunyi Medical University, Zunyi 563003, China
College of Laboratory Medicine, Zunyi Medical University, Zunyi 563006, China
Yan Renqing
Department of Laboratory Medicine, Affiliated Hospital of Zunyi Medical University, Zunyi 563003, China
College of Laboratory Medicine, Zunyi Medical University, Zunyi 563006, China
Zhang Wei
Jiangsu Qlife Medical Technology Group Co., LTD., Nanjing 211899, China
Yan Shengkai
Department of Laboratory Medicine, Affiliated Hospital of Zunyi Medical University, Zunyi 563003, China
College of Laboratory Medicine, Zunyi Medical University, Zunyi 563006, China
