整体式C18高效液相色谱柱用于在盐酸毛果芸香碱降解产物存在下快速分析该药物的评估。

Evaluation of monolithic C18 HPLC columns for the fast analysis of pilocarpine hydrochloride in the presence of its degradation products.

作者信息

El-Deeb S, Schepers U, Wätzig H

机构信息

Institute of Pharmaceutical Chemistry, TU Braunschweig, Germany.

出版信息

Pharmazie. 2006 Sep;61(9):751-6.

PMID:17020149

Abstract

Monolithic Performance C18 HPLC columns (Chromolith Performance RP-18e, Merck) were applied for the determination of pilocarpine hydrochloride in the presence of its degradation products isopilocarpine, pilocarpic acid and isopilocarpic acid. The method was validated using a set of six monolithic columns and compared to a conventional C18 column. The separation of pilocarpine from its degradation products was investigated on monolithic columns at different flow rates from 1 to 9 ml/min. Superior resolution was obtained using monolithic columns over the conventional C18 column at the same flow rate of 1 ml/min with a total run time of 9 min compared to 13.5 min for the conventional C18 column. Comparable resolution to that obtained in the C18 column (but with better peak symmetry) was obtained at a flow rate of 4 ml/min, although the total run time was reduced to 3.5 min. The precision for both retention time and peak area was investigated over a wide concentration range and found to be equal, or slightly better on Chromolith Performance compared to the conventional column. The overall RSDs% ranged from 0.5% to 1.16% for the conventional column, while for monolithic columns ranged from 0.38% to 0.87% at a flow rate of 1 ml/min and from 0.38% to 0.89% at a flow rate of 4 ml/min. Monolithic column to column reproducibility (n = 6) was measured. The RSDs% ranged from 0.34% to 0.68% for retention time and from 0.3% to 0.94% for peak areas. The detection and quantitation limits on monolithic columns at both flow rates (1 and 4 ml/min) were found to be 0.17 microg/ml and 0.5 microg/ml, compared to 0.31 microg/ml and 1 microg/ml on the conventional column. Monolithic silica rods have also shown the advantage of reducing the time to wash and to re-equilibrate the column. The method showed good linearity and recovery and was found to be suitable for the analysis of pilocarpine hydrochloride formulations.

摘要

整体式高效液相色谱C18柱（默克公司的Chromolith Performance RP - 18e）用于在其降解产物异毛果芸香碱、毛果芸香酸和异毛果芸香酸存在的情况下测定盐酸毛果芸香碱。该方法使用一组六根整体式柱进行了验证，并与传统C18柱进行了比较。在1至9 ml/min的不同流速下，研究了整体式柱上毛果芸香碱与其降解产物的分离情况。在相同流速1 ml/min下，使用整体式柱比传统C18柱获得了更高的分离度，整体式柱的总运行时间为9分钟，而传统C18柱为13.5分钟。在流速为4 ml/min时，获得了与C18柱相当的分离度（但峰形更好），尽管总运行时间缩短至3.5分钟。在较宽的浓度范围内研究了保留时间和峰面积的精密度，发现与传统柱相比，Chromolith Performance柱的精密度相同或略好。传统柱的总体相对标准偏差（RSDs%）范围为0.5%至1.16%，而整体式柱在1 ml/min流速下的RSDs%范围为0.38%至0.87%，在4 ml/min流速下为0.38%至0.89%。测量了整体式柱之间的重现性（n = 6）。保留时间的RSDs%范围为0.34%至0.68%，峰面积的RSDs%范围为0.3%至0.94%。发现在两种流速（1和4 ml/min）下整体式柱的检测限和定量限分别为0.17 μg/ml和0.5 μg/ml，而传统柱分别为0.31 μg/ml和1 μg/ml。整体式硅胶柱还显示出减少柱清洗和重新平衡时间的优势。该方法显示出良好的线性和回收率，适用于盐酸毛果芸香碱制剂的分析。