Mostafa Gamal A E, Hefnawy Mohammed M, El-Majed Abdulrahman
King Saud University, College of Pharmacy, Pharmaceutical Chemistry Department, PO Box 2457, Riyadh 11451, Saudi Arabia.
J AOAC Int. 2009 May-Jun;92(3):824-9.
Enantiomers of clenbuterol were separated by a new HPLC method on a chiral column. Enantiomeric resolution was achieved on a vancomycyin macrocyclic antibiotic chiral stationary phase known as chirobiotic V with UV detection at 247 nm. The polar ionic mobile phase consisting of methanol-triethylamine-glacial acetic acid (100 + 0.05 + 0.025, v/v/v), was used at a flow rate of 1.0 mL/min. The method was validated for linearity, accuracy, precision, and robustness. Standard linear calibration curves were established for the R-(-) and S-(+) enantiomers over the range of 0.2-20 microg/mL, and an average recovery of 98.0% and a mean relative standard deviation of 1.5% were obtained at 5.0 microg/mL. The lower limit of detection was 0.05 microg/mL for each enantiomer. The mean recovery for R-(-) and S-(+)-clenbuterol enantiomers from plasma was 91.0-97.0% at 0.20-20 microg/mL. The method was successfully used to identify and quantify the clenbuterol enantiomers in human plasma.
采用一种新的高效液相色谱法在手性柱上分离克伦特罗的对映体。在一种名为手性生物V的万古霉素大环抗生素手性固定相上实现了对映体分离，并在247nm处进行紫外检测。使用由甲醇 - 三乙胺 - 冰醋酸（100 + 0.05 + 0.025，v/v/v）组成的极性离子流动相，流速为1.0 mL/min。该方法针对线性、准确度、精密度和稳健性进行了验证。在0.2 - 20μg/mL范围内建立了R - (-)和S - (+)对映体的标准线性校准曲线，在5.0μg/mL时平均回收率为98.0%，平均相对标准偏差为1.5%。每种对映体的检测下限为0.05μg/mL。在0.20 - 20μg/mL时，血浆中R - (-)和S - (+)-克伦特罗对映体的平均回收率为91.0 - 97.0%。该方法成功用于鉴定和定量人血浆中的克伦特罗对映体。
