Nemutlu E, Celebier M, Uyar B, Altinöz S
Department of Analytical Chemistry, Faculty of Pharmacy, University of Hacettepe, 06100 Ankara, Turkey.
J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Jul 1;854(1-2):35-42. doi: 10.1016/j.jchromb.2007.03.050. Epub 2007 Apr 12.
An efficient and reliable micellar electrokinetic capillary chromatography (MEKC) method has been developed for the simultaneous determination of isoniazid (ISO) and pyridoxine hydrochloride (PYR) in pharmaceutical formulations. A chemometric two level full factorial design approach was used to search for the optimum conditions of separation. Three parameters were selected for this study: the buffer pH, the buffer concentration and sodium dodecyl sulphate (SDS) concentrations. Resolution, peak symmetry and analysis time were established as response. The two analytes were separated within 6 min with the optimized conditions: 50 mM borate buffer, 25 mM SDS pH 7.8, 35 degrees C, at 50 mbar 4s injection and 30 kV by using a fused silica capillary (72 cm effective length, 50 microm i.d.). The detection wavelength was set to 205 nm. Meloxicam was used as internal standard. The method was validated with respect to stability, linearity range, limit of quantitation and detection, precision, accuracy, specificity and robustness. The detection limits of the method were 1.0 microg mL(-1) for ISO and 0.40 microg mL(-1) for PYR and the method was linear at least in the range of 3.0-100 microg mL(-1) for ISO and 1.0-100 microg mL(-1) for PYR with excellent correlation coefficients (0.9995 for ISO and 0.9998 for PYR). Relative standard deviations (R.S.D.s) of the described method ranged between 0.54 and 2.27% for intra-day precision and between 0.65 and 2.69% for inter-day precision. The developed method was applied to the tablet form of ISO and PYR-containing the pharmaceutical preparations and the data were compared with obtained from the standard addition method. No statistically significant difference was found.
已开发出一种高效可靠的胶束电动毛细管色谱法（MEKC），用于同时测定药物制剂中的异烟肼（ISO）和盐酸吡哆醇（PYR）。采用化学计量学二级全因子设计方法来寻找分离的最佳条件。本研究选择了三个参数：缓冲液pH值、缓冲液浓度和十二烷基硫酸钠（SDS）浓度。将分离度、峰对称性和分析时间确定为响应指标。在优化条件下，两种分析物在6分钟内分离：50 mM硼酸盐缓冲液，25 mM SDS，pH 7.8，35℃，50 mbar下4秒进样，30 kV，使用熔融石英毛细管（有效长度72 cm，内径50 µm）。检测波长设定为205 nm。美洛昔康用作内标。该方法在稳定性、线性范围、定量限和检测限、精密度、准确度、特异性和稳健性方面进行了验证。该方法的检测限为ISO 1.0 μg mL⁻¹、PYR 0.40 μg mL⁻¹，该方法至少在ISO 3.0 - 100 μg mL⁻¹和PYR 1.0 - 100 μg mL⁻¹范围内呈线性，相关系数良好（ISO为0.9995，PYR为0.9998）。所述方法的日内精密度相对标准偏差（R.S.D.s）在0.54%至2.27%之间，日间精密度在0.65%至2.69%之间。所开发的方法应用于含ISO和PYR的药物制剂片剂形式，并将数据与标准加入法获得的数据进行比较。未发现统计学上的显著差异。
