摘要
目的:利用代谢组学代谢靶标分析技术,建立一种定量检测人血清中溶血磷脂酰胆碱(LPC)类物质的方法。方法:以高效液相色谱与串联质谱联用(HPLC-MS/MS)分析平台,利血平为内标物,构建定量检测LPC14∶0、LPC 15∶0、LPC 16∶0、LPC 17∶0和LPC 18∶0系列物质的检测方法。绘制每种目标LPC对应的标准曲线,分析其相关系数r值,并进一步评估检测平台的灵敏度检测下限(LOD)和线性范围。通过加样回收试验验证检测平台的准确度(加样回收率)和精密度(RSD),并进一步分析该平台在检测人血清标本的适用性。在此基础上,利用该HPLC-MS/MS平台分析了LPC 14∶0～LPC 18∶0 5个LPCs在乙肝相关性肝细胞肝癌(HBV-HCC)患者血清中的浓度。结果:本研究建立了基于HPLC-MS/MS的LPC检测平台,该平台检测稳定性和准确性较高。LPC 14∶0～LPC 18∶0 5个物质的检测下限分别是0.027、0.120、0.320、0.059、0.072μg·mL^(-1);标准曲线方程分别为Y=0.725X-0.002 64、Y=0.212X-0.025 6、Y=1.22X+1.84、Y=0.861X-0.015 5和Y=0.813X+0.558,相关系数分别为0.997 6、0.999 2、0.992 7、0.998 0和0.994 3,测量结果准确度和精密度均控制在85%～115%范围内。利用HPLC-MS/MS平台测定的5个LPC类物质,对不同巴塞罗那期HBV-HCC患者具备一定的鉴别能力。结论:本研究利用HPLC-MS/MS分析平台,成功建立了一个稳定、灵敏而可靠的定量检测人血清中LPC 14∶0～LPC 18∶0 5个目标LPC类物质的检测方法,适合临床推广。
Objective:To establish a quantitative detection platform for the identification of lysophosphatidylcholine(LPC)substances in human serum by using targeted metabolomics techniques.Methods:The analytical platform was developed based on a high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS),which using reserpine as the internal standard to quantitatively detect LPC substances,including LPC 14∶0,LPC 15∶0,LPC 16∶0,LPC 17∶0 and LPC 18∶0.The standard curve of each LPC was plotted and the linear correlation coefficient was analyzed.The limits of detection(LODs)and the linearity were evaluated.The accuracy(spiked recovery rate)and precision(RSD)of the detection platform were verified by spiked recovery experiments.The applicability of HPLC-MS/MS platform in human serum was analyzed,as well.In addition,the concentrations of five types of LPCs(LPC 14∶0-LPC 18∶0)in patients with hepatitis B-related hepatocellular carcinoma(HBV-HCC)were validated by the current HPLC-MS/MS platform.Results:A LPC detection platform based on HPLC-MS/MS was established,which had high stability and accuracy.The LODs of the five substances LPC 14∶0 to LPC 18∶0 were 0.027,0.120,0.320,0.059 and 0.072 μg·mL-1,respectively.The standard curves were:Y=0.725 X-0.002 64,Y=0.212 X-0.025 6,Y=1.22 X+1.84,Y=0.861 X-0.015 5 and Y=0.813 X+0.558,respectively.The correlation coefficients were 0.997 6,0.999 2,0.992 7,0.998 0 and 0.994 3,respectively.In the meantime,the accuracy and precision of the measurement were controlled within the range of 85%-115%.The five types of LPCs determined by HPLC-MS/MS platform could be applied to differentiate HBV-HCC patients with different stages of Barcelona.Conclusions:In the study,a stable,sensitive and reliable quantitative detection platform was successfully established based on HPLC-MS/MS.The platform can quantitatively detect five types of target LPCs(LPC 14∶0 to LPC 18∶0)in human serum,which is suitable for promotion in the clinical study.
作者
张磊
武玉晶
范志娟
康华
王宇凡
刘树业
单忠强
ZHANG Lei;WU Yu-jing;FAN Zhi-juan;KANG Hua;WANG Yu-fan;LIU Shu-ye;SHAN Zhong-qiang(College of Chemical Engineering,Tianjin University,Tianjin 300072,China;Department of Inspection,Tianjin Third Central Hospital,Tianjin Artificial Cell Key Laboratory,Ministry of Health Artificial Cell Engineering Technology Research Center,Tianjin 300170,China)
出处
《药物分析杂志》
CAS
CSCD
北大核心
2019年第2期272-279,共8页
Chinese Journal of Pharmaceutical Analysis
基金
天津市自然科学基金(青年项目)<慢性噬肝性病毒相关原发肝癌个性化诊疗代谢标志物研究>(No.16JCQNJC11600)
作者简介
通信作者:刘树业,Tel:15522242777,E-mail:lshye@163.com通信作者:单忠强,Tel:13612032260,E-mail:18622431776@163.com第一作者:张磊,Tel:13752068733,E-mail:rikkichang12345@163.com。
