Rosseel M T, Vermeulen A M, Belpaire F M
Heymans Institute of Pharmacology, University of Ghent Medical School, Belgium.
J Chromatogr. 1991 Jul 17;568(1):239-45. doi: 10.1016/0378-4347(91)80359-k.
A sensitive high-performance liquid chromatographic method for the determination of the enantiomers of atenolol in rat plasma has been developed. Racemic atenolol and practolol (internal standard) were extracted from alkalinized plasma (pH 12) into dichloromethane containing 3% (v/v) heptafluoro-1-butanol, and the organic layer was evaporated. The samples were derivatized with (+)-1-(9-fluorenyl)ethyl chloroformate at pH 8.5 for 30 min. After removal of excess reagent, the diastereomers were extracted into dichloromethane. The diastereomers were separated on a Microspher C18 column (3 microns) with a mobile phase of acetonitrile-sodium acetate buffer (0.01 M, pH 7) (50:50, v/v) at a flow-rate of 0.8 ml/min. Fluorescence detection (lambda ex = 227 nm, lambda em = 310 nm) was used. When 100 microliters of plasma were used, the quantitation limit was 10 ng/ml for the atenolol enantiomers. The assay was applied to measure concentrations of atenolol enantiomers in plasma after intravenous administration of racemic atenolol to rats.
已开发出一种灵敏的高效液相色谱法，用于测定大鼠血浆中阿替洛尔对映体。将外消旋阿替洛尔和醋丁洛尔（内标）从碱化血浆（pH 12）中萃取到含有3%（v/v）七氟-1-丁醇的二氯甲烷中，然后将有机层蒸发。样品在pH 8.5条件下用（+）-1-（9-芴基）氯甲酸乙酯衍生化30分钟。去除过量试剂后，将非对映异构体萃取到二氯甲烷中。非对映异构体在Microspher C18柱（3微米）上分离，流动相为乙腈-醋酸钠缓冲液（0.01 M，pH 7）（50:50，v/v），流速为0.8 ml/min。采用荧光检测（激发波长λex = 227 nm，发射波长λem = 310 nm）。当使用100微升血浆时，阿替洛尔对映体的定量限为10 ng/ml。该方法用于测定给大鼠静脉注射外消旋阿替洛尔后血浆中阿替洛尔对映体的浓度。
