摘要
以R(-)-4-(N,N-Dimethylaminosulfonyl)-7-(3-isothiocyanatopyrrolidino)-2,1,3-benzoxadiazole(R(-)-DBD-PyNCS)作为手性荧光衍生化试剂,用反相高效液相色谱法(RP-HPLC)定量分析多巴胺.衍生化反应是在含有7%吡啶溶剂中,75℃温度条件下,进行40min,生成具有荧光特性的多巴胺衍生物(λex=460 nm,λem=550nm).生成的衍生物在流动相为乙腈-水(体积比35:65),流速为1.0mL/min,在DiamonsilTM C8(150 mm×4.6mm,i.d.,5 μm)色谱柱上保留时间约为15min.多巴胺衍生物在60～1 200ng/mL时与峰面积呈良好的线性关系(r=0.999 6),多巴胺浓度为1 mmol/L时,峰面积测定结果的相对标准偏差(RSD)为2.4%,检出限为12 ng/L(S/N=3).检测了盐酸多巴胺注射液中多巴胺的含量.
Catecholamines,suchas epinephrine and dopamine,are importantsubstances inthe fields ofbiologiˉcal and clinical chemistry.This paper describes a new method developed to separate dopamine derivatized with fluorescent chiral reagent,R(-)-DBD-PyNCS:R(-)-4-(N,N-dimethylaminosulfonyl)-7-(3-iso- thiocyanatopyrrolidino)-2,1,3-benzoxadiazole,with reversed-phase high-performance liquid chromatography coupled witha Diamonsil TM C 18 column(150mm×4.6mm,i.d.,5μm)and a mobile phase ofwater-acetoniˉtrile(65∶35byvolume).Afluorescentdetectorwithanexcitationwavelengthof460nmand anemissionwaveˉlength of550nmwas employed for detection.The column was eluted at40℃with a flowrate of1mL/min.Fluorescent chiral reagent(30mmol/L,10μL)reacted with dopamine(1mmol/L,10μL),and10μL of20%pyridine acetonitrile solution was added.The resulting mixture was stirred for1min and allowed to react at75℃for40min.A10μLaliquotofthe derivatives was injected directlyinto the chromatograph.The retenˉtion time of dopamine was15min.The linear range of calibration was60～1200ng/mL(r=0.9996).The relative standard deviation(RSD)was2.4%,and the detection limits of dopamine was12ng(S/N=3).
出处
《分析测试学报》
CAS
CSCD
北大核心
2005年第2期73-75,共3页
Journal of Instrumental Analysis
关键词
手性荧光衍生化试剂
高效液相色谱法
多巴胺
Chiral fluorescent reagent
High-performance liquid chromatography
Epinephrine enantiomers
