Chocholous Petr, Satínský Dalibor, Solich Petr
Department of Analytical Chemistry, Faculty of Pharmacy, Charles University, Heyrovského 1203, Hradec Králové 50005, Czech Republic.
Talanta. 2006 Sep 15;70(2):408-13. doi: 10.1016/j.talanta.2006.02.065. Epub 2006 Apr 11.
Fast simultaneous determination of naphazoline nitrate and methylparaben in pharmaceuticals using separation method based on a novel reversed-phase sequential injection chromatography (SIC) is described in this contribution as an alternative to classical HPLC. A Chromolithtrade mark Flash RP-18e (25mmx4.6mm) column (Merck((R)), Germany) and a FIAlab((R)) 3000 system (USA) with a six-port selection valve and 5.0ml syringe pump were used for sequential injection chromatographic separations in our study. The mobile phase used was methanol/water (40:65, v/v), pH 5.2 adjusted with triethylamine 0.8mulml(-1) and acetic acid, at flow rate 0.9mlmin(-1). UV detection provided by DAD detector and two wavelengths were simultaneously monitored for increasing sensitivity of determination. Detector was set up at 220nm for naphazoline nitrate and 256nm for methylparaben and ethylparaben (IS). There is no necessity to use pre-adjustment of sample of nasal drops (only dilution with mobile phase) so the time of the whole analysis is very short. The validation parameters have shown good results: linearity of determination for both components (naphazoline nitrate and methylparaben), correlation coefficient >0.999; repeatability of determination (R.S.D.) in the range 0.5-1.6% at three different concentration levels, detection limits 0.02mugml(-1) (naphazoline nitrate) and 0.20mugml(-1) (methylparaben and ethylparaben), and recovery from the pharmaceutical preparations in the range 100.06-102.55%. The chromatographic resolution between peaks of compounds was more than 4.0 and analysis time was less than 4min under the optimal conditions. The advantages and drawbacks of SIC against classical HPLC are discussed showing that SIC can be an advantageous alternative in many cases.
本文介绍了一种基于新型反相顺序注射色谱法(SIC)的分离方法,用于快速同时测定药品中的硝酸萘甲唑啉和对羟基苯甲酸甲酯,作为经典高效液相色谱法的替代方法。在我们的研究中,使用了Chromolith™ Flash RP - 18e(25mm×4.6mm)柱(德国默克公司)和带有六通选择阀及5.0ml注射泵的FIAlab™ 3000系统(美国)进行顺序注射色谱分离。所用流动相为甲醇/水(40:65,v/v),用0.8μl/ml三乙胺和乙酸调节pH至5.2,流速为0.9ml/min。由二极管阵列检测器(DAD)提供紫外检测,并同时监测两个波长以提高测定灵敏度。检测器设置为硝酸萘甲唑啉在220nm,对羟基苯甲酸甲酯和对羟基苯甲酸乙酯(内标)在256nm。滴鼻剂样品无需预先调节(仅用流动相稀释),因此整个分析时间非常短。验证参数显示出良好的结果:两种成分(硝酸萘甲唑啉和对羟基苯甲酸甲酯)测定的线性,相关系数>0.999;在三个不同浓度水平下测定的重复性(相对标准偏差)在0.5 - 1.6%范围内,检测限为0.02μg/ml(硝酸萘甲唑啉)和0.20μg/ml(对羟基苯甲酸甲酯和对羟基苯甲酸乙酯),药物制剂中的回收率在100.06 - 102.55%范围内。在最佳条件下,化合物峰之间的色谱分辨率大于4.0,分析时间小于4分钟。讨论了SIC相对于经典HPLC的优缺点,表明SIC在许多情况下可能是一种有利的替代方法。