Rao R Nageswara, Shinde Dhananjay D
Analytical Chemistry Division, Discovery Laboratory, Indian Institute of Chemical Technology, Tarnaka, Hyderabad, India.
Biomed Chromatogr. 2009 Aug;23(8):811-6. doi: 10.1002/bmc.1190.
A simple and rapid normal-phase HPLC method for enantiospecific separation of a psychostimulant, adrafinil (ADL), and its metabolite modafinil (MDL) in rat serum and urine was developed. The separation was accomplished on a normal-phase polysaccharide stationary phase Chiralcel OJ-H using n-hexane-ethanol (62:38 v/v) as a mobile phase at a flow rate of 1.0 mL/min. Detection was carried out at 225 nm using a photo diode array (PDA) detector. The elution order of the enantiomers was determined by a polarimeter connected in series with the PDA. ADL and its metabolite were recovered from rat serum and urine by solid phase extraction using Oasis HLB cartridges and the mean recoveries were >or=80%. The enantiomers were eluted within 15 min without any interference from endogenous substances. The calibration curves were linear (r(2) > 0.998) in the concentration range of 1.20-500 microg/mL for ADL and MDL. The assay was specific, accurate, precise and reproducible (intra- and inter-day precisions RSDs <7.2%). ADL in rat serum was stable over three freeze-thaw cycles at ambient temperature for 4 h. The method was successfully applied to pharmacokinetic studies of adrafinil after an oral administration to rats.
建立了一种简单快速的正相高效液相色谱法，用于对大鼠血清和尿液中的精神兴奋药阿屈非尼（ADL）及其代谢产物莫达非尼（MDL）进行对映体特异性分离。分离在正相多糖固定相Chiralcel OJ-H上进行，以正己烷-乙醇（62:38 v/v）为流动相，流速为1.0 mL/min。使用光电二极管阵列（PDA）检测器在225 nm波长处进行检测。对映体的洗脱顺序通过与PDA串联的旋光仪确定。使用Oasis HLB柱通过固相萃取从大鼠血清和尿液中回收ADL及其代谢产物，平均回收率≥80%。对映体在15分钟内洗脱，无内源性物质干扰。ADL和MDL在1.20 - 500 μg/mL浓度范围内校准曲线呈线性（r(2) > 0.998）。该测定法具有特异性、准确性、精密度和重现性（日内和日间精密度RSDs < 7.2%）。大鼠血清中的ADL在室温下4小时内经过三个冻融循环仍稳定。该方法成功应用于大鼠口服阿屈非尼后的药代动力学研究。
