LC-MS/MS法测定抑郁症患者晨尿中9种氨基酸类内源性物质
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篇名: LC-MS/MS法测定抑郁症患者晨尿中9种氨基酸类内源性物质
TITLE:
摘要:
摘 要 目的建立测定人体尿液中5-羟基吲哚乙酸谷氨酰胺马尿酸庚二酸脯氨酸酪氨酸色氨酸酪胺缬氨酸浓度的方 方法采集抑郁症患者的晨尿加入内标可的松以乙腈处理后取上清液进行浓缩采用液相色谱-质谱串联法LC-MS/MS行分析色谱柱为XTerra RP18流动相A相为0.1乙酸-流动相B相为0.1乙酸-乙腈梯度洗脱);柱温为40 流速为0.45 mL/min采用电喷雾离子源ESI),以多反应监测方式MRM进行定量分析
5-羟基吲哚乙酸谷氨酰胺马尿酸庚二酸脯氨酸酪氨酸色氨酸酪胺缬氨酸和内标可的松在正离子模式下定量分析离子对分别为质荷比m/z192.2146.1147.2130.0180.1105.1161.1125.2116.170.2205.2188.2138.2121.1182.0123.0118.272.1361.2163.0结果5-羟基吲哚 乙酸谷氨酰胺缬氨酸庚二酸脯氨酸酪胺的检测质量浓度范围为10.00~3 200 ng/mLr=0.993 8~0.998 9n=6),定量下限
10 ng/mL色氨酸酪氨酸马尿酸的检测质量浓度范围为1 600~51 200 ng/mLr=0.999 2~0.999 7n=6),定量下限为1 600 ng/mL准确度试验结果为 86.29~98.65n=6),日内日间精密度试验的 RSD 均不高于 14.65n=6),基质效应的 CV 6.18~14.37n=6),提取回收率为86.21~98.14n=6),稳定性试验的RE<14.71n=3~6)。结论该方法灵敏可用于测定人体内上述9种物质的浓度
ABSTRACT:
ABSTRACT OBJECTIVETo establish a method for the determination of 5-hydroxyindole acetic acid5-HIAA),glutamine
hippuratepimelateprolinetryptophantyraminetyrosine and valine in human urine. METHODSMorning urine samples were collected from depression patients. The sample was extracted with acetonitrile after addition of internal standard cortisone. LC-MS/ MS method was adopted. The determination was performed on XTerra RP18 column with mobile phase consisted of 0.1acetic
acid-water as mobile phase A and 0.1acetic acid-acetonitrile as mobile phase Bgradient elution. The column temperature was
set at 40 and flow rate was 0.45 mL/min. Electrospray ion sourceESIwas used for quantitative analysis by multiple reaction monitoring MRM. The precursor/product ion transitions m/zwere monitored at m/z 192.2146.1m/z 147.2130.0m/z 180.1105.1m/z 161.1125.2m/z 116.170.2m/z 205.2188.2m/z 138.2121.1m/z 182.0123.0m/z 118.272.1 and m/z 361.2163.0+ion-modefor 5-HIAAglutaminehippuratepimelateprolinetryptophantyraminetyrosinevaline and cortisonerespectively. RESULTSThe linear range of 5-HIAAglutaminepimelateprolinetyramine and valine were 10.00-3 200 ng/mLr=0.993 8-0.998 9n=6. The lower limit of quantification was 10 ng/mLthe linear range of hippuratetryptophan and tyrosine were 1 600-51 200 ng/mLr=0.999 2-0.999 7n=6. The lower limit of quantitation was 1 600 ng/mL. The results of accuracy tests were 86.29-98.65n=6. RSDs of intra-day and inter-day precision tests were no more than 14.65n=6CV of matrix effect were 6.18-14.37n=6. Extraction recovery rates were 86.21-98.14n=6. RE of stability tests were no more than 14.71n=3-6. CONCLUSIONSThe method is sensitiveaccurate and suitable for the determination of 9 substances in human.
期刊:
作者: 宋红艳,夏敬胜,纪刚剑,任章焘,张相林,柳 芳,陈 灵
AUTHORS: SONG Hongyan,XIA Jingsheng,JI Gangjian,REN Zhangtao,ZHANG Xianglin,LIU Fang,CHEN Ling
关键字: 抑郁症患者;尿液浓度;液相色谱-质谱串联法;氨基酸类物质
KEYWORDS: Depression patients;Urine concentration;LC-MS/MS;Amino acid substances
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