固相萃取-高效液相色谱-荧光法测定人体尿液中单胺类神经递质

Determination of monoamine neurotransmitters in human urine by solid phase extraction-high performance liquid chromatography-fluorescence detector

建立一种快速、准确测定人体尿液中肾上腺素、去甲肾上腺素、多巴胺及5-羟色胺4种单胺类神经递质的方法。尿液样品以Waters Oasis WCX固相萃取柱富集,依次用2 m L 20 mmol/L乙酸铵-甲醇溶液、2 m L甲醇淋洗杂质,最后用5 m L含2%甲酸的乙腈水溶液(85∶15,ν∶ν)提取被分析物。经Agilent C18色谱柱(250 mm×4.6 mm,5μm)分离,以甲醇与乙酸钠溶液的不同配比设置流动相梯度,在荧光检测激发波长280 nm,发射波长340 nm条件下,测定人体尿液中4种单胺类神经递质的含量。结果表明:4种单胺类神经递质的回收率为92.2%~104.8%,相对偏差为1.6%~3.1%,检出限为0.002~0.05μg/m L。该方法稳定、快速、简便,能满足人体尿液中多种单胺类神经递质的含量测定。

A rapid and accurate method for detecting pinephrine, norepinephrine, dopamine, serotonin hydrochloridein human urine was developed. The monoamine neurotransmitters were extracted from human urine samples with Waters Oasis WCX solid phase extraction(SPE) cartridges. Then 2 mL 20 mmol/L ammonium acetate -methanol solution, 2 mL methanol were used for eluting impurities. Finally,the analyte were extracted by 5 mL acetonitrile -water solution (85:15,淄:淄) containing 2% formic acid. The separation was performed on Agilent C18 capillary column (250mmx 4.6 mm,5 m) followed by different proportion of methanol and sodium acetate solutions gradient elution. Four monoamine neurotransmitters in human urine were detected by high performance liquid chromatography with fluorescence detection (excitation wavelength 280 nm, emission wavelength 340 nm). The results showed the average recoveries were between 92.2% and 104.8%, the relative standard deviations were between 1.6% and 3.1%,and the limits of detection were between 0.002 g/mL and 0.05 g/mL. The method used in this study is stable,fast and easy. The method has practical value for detecting monoamine neurotransmitters in human urine.