摘要
以R (- ) /S (+) 4 (N ,N dimethylaminosulfonyl) 7 (3 iso thiocyanatopyrrolidino) 2 ,1,3 benzoxadiazole(DBD PyNCS)为手性荧光衍生化试剂 ,用反相高效液相色谱法 (RP HPLC)对 DL 肾上腺素对映体进行分离。衍生化反应是在含有 7%吡啶的溶剂中 ,6 5℃温度条件下反应 35min ,生成具有荧光特性的肾上腺素非对映体衍生物(λex=4 6 0nm ,λem=5 5 0nm)。生成的非对映体衍生物在流动相为乙腈 水 (体积比为 2 8∶72 ) ,流速为 1 0mL/min时 ,在DiamonsilTMC18色谱柱 (15 0mm× 4 6mmi d ,5 μm)上分离度可达 2 6。肾上腺素对映体在 0 0 7g/L～ 0 5 0g/L时与峰面积呈良好的线性关系 (r =0 9995 ) ,峰面积测定结果的相对标准偏差为 4 3% (n =7) ,最低检测限为12ng/L(S/N =3)。
A new method was developed to separate epinephrine enantiomers derivatized with fluorescent chiral reagent, R (-)/S (+) 4 ( N,N dimethylaminosulfonyl) 7 (3 iso thiocyanatopyrro lidino) 2, 1, 3 benzoxadiazole (DBD PyNCS), by reversed phase high performance liquid chromatography, on a Diamonsil TM C 18 column(150 mm×4 6 mm i d , 5 μm), with a mobile phase of water acetonitrile(72∶28, volume ratio) and a fluorescent detector at the excitation wavelength of 460 nm and the emission wavelength of 550 nm. The column was eluted at 40 ℃, with a flow rate of 1 mL/min . At first, let the fluorescent chiral reagent(36 mmol/L, 10 μL) react with epinephrine enantiomers(1 mmol/L, 10 μL), and then 10 μL 20% pyridine acetonitrile solution was added. The resulting mixture was stirred for 1 min and kept at 65 ℃ for 35 min and then a 10 μL aqueous of the diastereomeric derivatives was injected directly into the chromatograph. The diastereomeric compounds were efficiently separated. The R s value was 2 6 with α of 1 07. The k ′s of L epinephrine and D epinephrine were 17 11 and 18 13 respectively. The retention times of L epinephrine and D epinephrine were 30 24 min and 31 94 min respectively. The linear range was 0 07 g/L-0 50 g/L ( r = 0 999?5 ). The RSD was 4 3%( n =7). The detection limit was 12 g/L( S/N =3).
出处
《色谱》
CAS
CSCD
北大核心
2002年第4期304-307,共4页
Chinese Journal of Chromatography
基金
教育部留学回国人员科研启动基金资助项目
关键词
分离
手性荧光衍生化试剂
反相高效液相色谱法
肾上腺素对映体
分析
fluorescent chiral derivatization reagent
reversed phase high performance liquid chromatography
epinephrine enantiomer
