A SENSITIVE AND A SIMPLE RP-HPLC METHOD DEVELOPMENT AND VERIFICATION FOR THE QUANTITATIVE ESTIMATION OF CHOLECALCIFEROLIN TABLETS

K. Subhashini, U. T. Naidu, A. A. Kumar, Rajesh Vooturi, N. Kumar
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Abstract

Objective: To develop a sensitive, simple, accurate, precise and linear Reverse Phase High-Performance Liquid Chromatographic (RP-HPLC) method and verify for the quantitative estimation (Assay) of Cholecalciferol in tablets. Methods: The optimized method uses a reverse phase column, Waters X-Bridge C8 (150 X 4.6 mm; 3.5μ), a mobile phase of Methanol: Acetonitrile: HPLC grade water/Milli-Q water in the proportion of 60:30:10 v/v/v, flow rate of 0.8 ml/min, injection volume of 100 µl, and detection wavelength of 265 nm using a UV/PDA detector. Results: The developed method gave Cholecalciferol eluting at about 9 min. Cholecalciferol exhibited linearity in the range 0.058-0.466 μg/ml. The precision is exemplified by a relative standard deviation of 1.40%. Percentage of individual recovery was found to be in the range of 97.0 and 103.0 during accuracy studies. Conclusion: A sensitive, simple, accurate, precise and linear RP-HPLC method was developed and verified for the quantitative estimation (Assay) of Cholecalciferol in tablets and hence this method can be explored for the analysis of Cholecalciferol in tablets/various dosage forms in various pharmaceutical industries.
建立并验证灵敏简便的 RP-HPLC 方法定量测定胆囊收缩素片
目的建立一种灵敏、简便、准确、精确和线性的反相高效液相色谱法(RP-HPLC),并验证其对片剂中胆钙化醇的定量估计(Assay)。 方法:优化方法采用反相色谱柱 Waters X-Bridge C8 (150 X 4.6 mm; 3.5μ),流动相为甲醇:流动相为甲醇:乙腈:HPLC 级水/Milli-Q 水,流速为 0.8 ml/min,进样量为 100 µl,紫外/PDA 检测器的检测波长为 265 nm。 结果表明所开发的方法可在约 9 分钟内洗脱出胆钙化醇。胆钙化醇在 0.058-0.466 μg/ml 范围内呈线性关系。相对标准偏差为 1.40%。在准确度研究中发现,单个回收率在 97.0 和 103.0 之间。 结论该方法灵敏、简便、准确、精密、线性,可用于片剂中胆钙化醇的定量分析。
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