Hefnawy Mohamed M, Sultan Maha A, Al-Shehri Mona M
Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, PO Box 2457, Riyadh 11451, Saudi Arabia.
J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Sep 1;856(1-2):328-36. doi: 10.1016/j.jchromb.2007.06.021. Epub 2007 Jun 29.
A sensitive and selective high-performance liquid chromatographic (HPLC) method has been developed for the simultaneous determination of bufuralol enantiomers in plasma and pharmaceutical formulations. Enantiomeric resolution was achieved on a vancomycin macrocyclic antibiotic chiral stationary phase (CSP) known as Chirobiotic V with UV detection set at 254 nm. The polar ionic mobile phase (PIM) consisting of methanol-glacial acetic acid-triethylamine (100:0.015:0.010, v/v/v) has been used at a flow rate of 0.5 ml/min. The method is highly specific where other coformulated compounds did not interfere. The stability of bufuralol enantiomers under different degrees of temperature was also studied. The results showed that the drug is stable for at least 7 days at 70 degrees C. The method was validated for its linearity, accuracy, precision and robustness. An experimental design was used during validation to evaluate method robustness. The calibration curves in plasma were linear over the range of 5-500 ng/ml for each enantiomer with detection limit of 2 ng/ml. The mean relative standard deviation (RSD) of the results of within-day precision and accuracy of the drug were <or=10%. There was no significant difference (p>0.05) between inter- and intra-day studies for each enantiomer which confirmed the reproducibility of the assay method. The mean extraction efficiency for S-(-)- and R-(+)-bufuralol from plasma was in the range 97-102% at 15-400 ng/ml level for each enantiomer. The overall recoveries of bufuralol enantiomers from pharmaceutical formulations was in the range 99.6-102.2% with %RSD ranging from 1.06 to 1.16%. The assay method proved to be suitable as chiral quality control for bufuralol formulations by HPLC and for therapeutic drug monitoring.
已开发出一种灵敏且具选择性的高效液相色谱（HPLC）方法，用于同时测定血浆和药物制剂中的布呋洛尔对映体。在一种名为Chirobiotic V的万古霉素大环抗生素手性固定相（CSP）上实现了对映体分离，紫外检测波长设定为254 nm。由甲醇 - 冰醋酸 - 三乙胺（100:0.015:0.010，v/v/v）组成的极性离子流动相（PIM）以0.5 ml/min的流速使用。该方法具有高度特异性，其他共配制的化合物不产生干扰。还研究了布呋洛尔对映体在不同温度下的稳定性。结果表明，该药物在70℃下至少稳定7天。该方法在线性、准确性、精密度和稳健性方面得到了验证。在验证过程中采用实验设计来评估方法的稳健性。血浆中的校准曲线在每种对映体5 - 500 ng/ml范围内呈线性，检测限为2 ng/ml。该药物日内精密度和准确性结果的平均相对标准偏差（RSD）≤10%。每种对映体的日间和日内研究之间无显著差异（p>0.05），这证实了该测定方法的可重复性。在15 - 400 ng/ml水平下，S - (-)-和R - (+)-布呋洛尔从血浆中的平均提取效率在97 - 102%范围内。布呋洛尔对映体从药物制剂中的总回收率在99.6 - 102.2%范围内，%RSD在1.06至1.16%之间。该测定方法被证明适用于通过HPLC对布呋洛尔制剂进行手性质量控制以及治疗药物监测。
