Kumar G Pradeep, Srivastava Vishal, Khandelwal Kiran, Kumar Rajesh, Hiriyanna S G, Kumar Ajay, Kumar Pramod
Analytical Research Department, Micro Labs, Bommasandra, Bangalore.
API R&D Centre, Micro Labs, Bommasandra, Bangalore.
Chirality. 2016 Sep;28(9):628-32. doi: 10.1002/chir.22626.
Besifloxacin is a unique chiral broad-spectrum flouroquinolone used in the treatment of bacterial conjunctivitis. R-form of besifloxacin hydrochloride shows higher antibacterial activity as compared to the S-isomer. Therefore, it is necessary to establish chiral purity. To establish chiral purity a high-performance liquid chromatography (HPLC) method for determination of R-besifloxacin and S-besifloxacin (BES impurity A) was developed and validated for in-process quality control and stability studies. The analytical performance parameters such as linearity, precision, accuracy, specificity, limit of detection (LOD), and lower limit of quantification (LOQ) were determined according to International Council for Harmonization ICH Q2(R1) guidelines. HPLC separation was achieved on Chiralpak AD-H (250 x 4.6 mm, 5 μm) column using n-heptane: ethanol: ethylenediamine: acetic acid (800:200:0.5:0.5) (v/v/v/v) as the mobile phase in an isocratic elution. The eluents were monitored by UV/Visible detector at 290 nm. The resolution between S-isomer and besifloxacin hydrochloride was more than 2.0. Based on a signal-to-noise ratio of 3 and 10 the LOD of besifloxacin was 0.30 μg/mL, while the LOQ was 0.90 μg/mL. The calibration curves were linear in the range of 0.9-7.5 μg/mL. Precision of the method was established within the acceptable range. The method was suitable for the quality control enantiomeric impurity in besifloxacin hydrochloride. Chirality 28:628-632, 2016. © 2016 Wiley Periodicals, Inc.
贝西沙星是一种独特的手性广谱氟喹诺酮类药物，用于治疗细菌性结膜炎。与S-异构体相比，盐酸贝西沙星的R-型显示出更高的抗菌活性。因此，有必要确定其手性纯度。为了确定手性纯度，开发了一种用于测定R-贝西沙星和S-贝西沙星（BES杂质A）的高效液相色谱（HPLC）方法，并对其进行了验证，用于过程质量控制和稳定性研究。根据国际协调理事会ICH Q2(R1)指南确定了线性、精密度、准确度、专属性、检测限（LOD）和定量下限（LOQ）等分析性能参数。使用正庚烷：乙醇：乙二胺：乙酸（800:200:0.5:0.5）（v/v/v/v）作为流动相，在Chiralpak AD-H（250×4.6 mm，5μm）柱上进行等度洗脱，实现HPLC分离。通过紫外/可见检测器在290 nm波长处监测洗脱液。S-异构体与盐酸贝西沙星之间的分离度大于2.0。基于信噪比为3和10，贝西沙星的检测限为0.30μg/mL，定量下限为0.90μg/mL。校准曲线在0.9 - 7.5μg/mL范围内呈线性。该方法的精密度在可接受范围内。该方法适用于盐酸贝西沙星中对映体杂质的质量控制。《手性》28:628 - 632，2016年。©2016威利期刊公司。
