高效液相色谱-串联质谱联用法测定人血清中去甲肾上腺素、多巴胺和5-羟色胺含量
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- 英文论文题名：Determination of norepinephrine,dopamine and 5-hydroxytryptamine in human serum by HPLC-MS/MS
- 论文作者：王泽民 ; 王玉文 ; 伊晶 ; 龙森 ; 余露山
- 英文论文作者：WANG Zemin ; WANG Yuwen ; YI jing ; LONG Sen ; YU lushan ; The Seventh hospital of Hangzhou ; Institute of materia Medica Pharmaceutical College Zhejiang University
- 年：2014
- 作者机构：杭州市第七人民医院;浙江大学药学院药物研究所;
- 论文关键词：血清 ; 神经递质 ; HPLC-MS/MS ; 含量
- 英文论文关键词：serum ; Neurotransmitters ; HPLC/MS/MS ; content
- 会议召开时间：2014-11-20
- 会议录名称：2014浙江省医学会精神病学分会学术年会、浙江省医师协会精神科医师分会第七届年会论文汇编
- 语种：中文
- 分类号：R969
- 学会代码：ZJKX
- 会议名称：2014浙江省医学会精神病学分会学术年会、浙江省医师协会精神科医师分会第七届年会
- 会议地点：中国浙江湖州
- 主办单位：浙江省医学会精神病学分会（The Society of Psychiatry ; ZJMA）、浙江省医师协会精神科医师分会
- 学会名称：浙江省科学技术协会
- 页数：7
- 文件大小：656k
- 原文格式：O
- 会议级别：地方
摘要
目的:建立高效液相色谱-串联质谱联用法(HPLC-MS/MS)测定人血清中3种神经递质含量的方法。方法:血清样品经液-液萃取后,以乙腈-水-甲酸(75:25:0.1,v/v/v)为流动相,尼莫地平为内标,采用ZORBAXExtend C18(2.1*100mm,3.5μm)柱分离。通过HPLC-MS/MS法,以选择离子监测(MRM)方式进行检测。结果:测定血清样品中去甲肾上腺素(NA)、多巴胺(DA)和5-羟色胺(5-HT)的线性范围分别为0.5-400 ng·mL~(-1),1-100 ng·mL~(-1)-1和1-100 ng·Ml~(-1)。NA,DA和5-HT的最低定量限分别为0.5 ng·mL~(-1)(RSD%=11.5%,n=5),1 ng·mL~(-1)(RSD%=6.1%,n=5)和1 ng·mL~(-1)(RSD%=9.3%,n=5)。NA,DA和5-HT的提取回收率分别为83.0%-87.5%,86.2%-93.7%和88.4%-93.2%。基质效应、日内日间精密度和准确度都符合生物样品测定要求。结论:本法操作简便、准确、灵敏度高,适用于血清中神经递质的测定和研究。
Objective:To establish a high performance liquid chromatography tandem mass spectrometry(HPLC-MS/MS) method for the determination of 3 kinds of neurotransmitter in human serum.Method:After liquid-liquid extraction,the samples of serum use acetonitrile-water-formic acid(75:25:0.1) as mobile phase,and use Nimodipine as the internal standard,with the method ZORBAX Extend C18(2.1*100 mm,3.5 μm) column to separation.By HPLC/MS/MS method,use HPLC/MS/MS method to test.Results:The linear range of noradrenaline(NA),dopamine(DA) and 5-hydroxytryptamine(5-HT) of the serum samples were 0.5-400 ng·mL-1,1-100 ng ·mL-1 and 1-100 ng ·mL-1.The minimum quantitative limit of NA,DA and 5-HT were 0.5 ng ·mL-l(RSD%=11.5%,n=5),1 ng ·mL-l(RSD%=6.1%,n=5),and 1 ng ·mL-l(RSD%=9.3%,n=5).Their extraction recovery rates were 83.0%-87.5%,86.2%-93.7%and 88.4%-93.2%.The matrix effect,within day and between day precision and accuracy rating meet the requirement of the determination of biological samples.Conclusion:This method is simple,accurate,high sensitivity,and is suitable for the determination and research of neurotransmitters in serum.
Objective:To establish a high performance liquid chromatography tandem mass spectrometry(HPLC-MS/MS) method for the determination of 3 kinds of neurotransmitter in human serum.Method:After liquid-liquid extraction,the samples of serum use acetonitrile-water-formic acid(75:25:0.1) as mobile phase,and use Nimodipine as the internal standard,with the method ZORBAX Extend C18(2.1*100 mm,3.5 μm) column to separation.By HPLC/MS/MS method,use HPLC/MS/MS method to test.Results:The linear range of noradrenaline(NA),dopamine(DA) and 5-hydroxytryptamine(5-HT) of the serum samples were 0.5-400 ng·mL-1,1-100 ng ·mL-1 and 1-100 ng ·mL-1.The minimum quantitative limit of NA,DA and 5-HT were 0.5 ng ·mL-l(RSD%=11.5%,n=5),1 ng ·mL-l(RSD%=6.1%,n=5),and 1 ng ·mL-l(RSD%=9.3%,n=5).Their extraction recovery rates were 83.0%-87.5%,86.2%-93.7%and 88.4%-93.2%.The matrix effect,within day and between day precision and accuracy rating meet the requirement of the determination of biological samples.Conclusion:This method is simple,accurate,high sensitivity,and is suitable for the determination and research of neurotransmitters in serum.
引文
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[6]林凌云,李慧,许崇涛.HPLC-荧光检测法测定大鼠脑组织中的单胺类神经递质及相关物质[J].华西药学杂志,2007,22(5):557-559.
[7]黄建国,徐幸民,印道春.高效液相色谱-紫外光检测法测定血清5-羟色胺[J].宁夏医学杂志2002.24(12):745-746.
[8]张璐璐,冯芳,叶小敏等,高效液相色谱-荧光法测定小鼠脑组织中单胺递质及其相关物质[J].中国药科大学学报,2010,41(4):367-371.
[2]Nozaki O,Kawamoto H,Moriyama H.Total free catecholamines assay by identification of its two functional groups and microflow injection chemiluminescence[J].Luminescence,1999,14(6):369-374.
[3]李永生,阎学安,邵福源.中枢神经递质与学习记忆的相关性研究进展[J].实用医药杂志,2006,23(7):864-866.
[4]王国强,张亚林,李萍,等.不同临床类型强迫症患者脑脊液单胺类神经递质对照研究[J].临床心身疾病杂志,2010,16(1):1-4.
[5]Barr LC,Goodman WK,Price LH,et al.The serotonin hypothesis of obsessive-compulsive disorder[J].Int Clin Psychopharmacol,1993,12(8):79-82.
[6]林凌云,李慧,许崇涛.HPLC-荧光检测法测定大鼠脑组织中的单胺类神经递质及相关物质[J].华西药学杂志,2007,22(5):557-559.
[7]黄建国,徐幸民,印道春.高效液相色谱-紫外光检测法测定血清5-羟色胺[J].宁夏医学杂志2002.24(12):745-746.
[8]张璐璐,冯芳,叶小敏等,高效液相色谱-荧光法测定小鼠脑组织中单胺递质及其相关物质[J].中国药科大学学报,2010,41(4):367-371.