Zhang Xiaoxiao, Gao Lei, Zhang Zunjian, Tian Yuan
Key Laboratory of Drug Quality Control&Pharmacovigilance (China Pharmaceutical University), Ministry of Education, Nanjing, China.
State Key Laboratory of Natural Medicine, China Pharmaceutical University, Nanjing, China.
J Pharm Anal. 2017 Oct;7(5):303-308. doi: 10.1016/j.jpha.2017.06.003. Epub 2017 Jun 13.
A high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS) method was established for the separation and determination of acetyl-glutamine enantiomers (acetyl-L-glutamine and acetyl-D-glutamine) simultaneously. Baseline separation was achieved on Chiralpak AD-H column (250 mm × 4.6 mm, 5 µm). -Hexane (containing 0.1% acetic acid) and ethanol (75:25, v/v) were used as mobile phase at a flow rate of 0.6 mL/min. The detection was operated in the negative ion mode with an ESI source. [M-H] 187.0540 for enantiomers and [M-H] 179.0240 for aspirin (IS) were selected as detecting ions. The linear range of the calibration curve for each enantiomer was 0.05-40 µg/mL. The precision of this method at concentrations of 0.5-20 µg/mL was within 7.23%, and the accuracy was 99.81%-107.81%. The precision at LOQ (0.05 µg/mL) was between 16.28% and 17.56%, which was poor than that at QC levels. The average extraction recovery was higher than 85% for both enantiomers at QC levels. The pharmacokinetics of enantiomers was found to be stereoselective. There was not chiral inversion in vivo or in vitro between enantiomers.
建立了一种高效液相色谱-质谱联用（HPLC-MS）方法，用于同时分离和测定乙酰谷氨酰胺对映体（乙酰-L-谷氨酰胺和乙酰-D-谷氨酰胺）。在Chiralpak AD-H柱（250 mm×4.6 mm，5 µm）上实现了基线分离。以含0.1%乙酸的正己烷和乙醇（75:25，v/v）为流动相，流速为0.6 mL/min。采用电喷雾离子源（ESI）在负离子模式下进行检测。选择对映体的[M-H] 187.0540和阿司匹林（内标，IS）的[M-H] 179.0240作为检测离子。各对映体校准曲线的线性范围为0.05 - 40 µg/mL。该方法在0.5 - 20 µg/mL浓度下的精密度在7.23%以内，准确度为99.81% - 107.81%。定量限（LOQ，0.05 µg/mL）处的精密度在16.28%至17.56%之间，低于质量控制（QC）水平的精密度。在QC水平下，两种对映体的平均提取回收率均高于85%。发现对映体的药代动力学具有立体选择性。对映体在体内和体外均未发生手性转化。
