Ranjan Om Prakash, Nayak Usha Y, Reddy Meka Sreenivasa, Dengale Swapnil J, Musmade Prashant B, Udupa Nayanabhirama
Department of Pharmaceutics, Manipal College of Pharmaceutical Sciences, Manipal University, Manipal 576104, Karnataka, India.
Department of Pharmaceutical Quality Assurance, Manipal College of Pharmaceutical Sciences, Manipal University, Manipal 576104, Karnataka, India.
J Young Pharm. 2013 Dec;5(4):133-8. doi: 10.1016/j.jyp.2013.10.006. Epub 2013 Nov 26.
To develop a liquid-liquid extraction based reverse phase liquid chromatography method for estimation of montelukast in rabbit plasma.
Chromatographic separation was carried out using Phenomenex Luna C18 column (250 mm × 4.6 mm × 5 μm) with mobile phase composed of ammonium acetate buffer (20 Mm), pH 5.5 and acetonitrile in 20:80, v/v ratio. The analyte was monitored with UV detector at 345 nm. The developed method was validated with respect to linearity, accuracy, precision, specificity and stability.
The peak area ratio of montelukast (MKS) to that of internal standard was used for the quantification of samples. Calibration curves were linear in the concentration range of 20-2000 ng mL(-1). The LOD and LLOQ of present method were found out to be 10 ng mL(-1) and 20 ng mL(-1) respectively. The intra-day and inter-day %CV values for MKS were below 6.06% and 8.43%. Intra-day and inter-day accuracies were within 95.81% and 110.90%, respectively. Extraction recoveries of drug from rabbit plasma were >66.47%.
A simple, alternative, reproducible and sensitive HPLC-UV method was developed for MKS that can be used in preclinical pharmacokinetics.
建立一种基于液-液萃取的反相液相色谱法，用于测定兔血浆中的孟鲁司特。
采用Phenomenex Luna C18柱（250 mm×4.6 mm×5μm）进行色谱分离，流动相由醋酸铵缓冲液（20 mM）、pH 5.5和乙腈按20:80（v/v）比例组成。用紫外检测器在345 nm波长处监测分析物。对所建立的方法进行线性、准确度、精密度、特异性和稳定性验证。
采用孟鲁司特（MKS）与内标物的峰面积比来定量样品。校准曲线在20 - 2000 ng mL⁻¹浓度范围内呈线性。本方法的检测限和定量限分别为10 ng mL⁻¹和20 ng mL⁻¹。MKS的日内和日间变异系数（%CV）值分别低于6.06%和8.43%。日内和日间准确度分别在95.81%和110.90%以内。从兔血浆中提取药物的回收率>66.47%。
建立了一种简单、可替代、可重现且灵敏的用于MKS的高效液相色谱-紫外检测法，可用于临床前药代动力学研究。
