HPLC梯度洗脱法测定氯替泼诺妥布霉素复方混悬滴眼液中氯替泼诺含量及有关物质

摘要

目的：：建立HPLC梯度法测定氯替泼诺妥布霉素复方混悬滴眼液中氯替泼诺含量及有关物质。方法：采用HPLC法,色谱柱：Inertsil ph苯基柱(250 mm ×4.6 mm,5μm)；流动相A：0.25%醋酸溶液-乙腈(80∶20)；流动相B：乙腈；流速：2.0 ml·min-1,进行线性梯度洗脱；检测波长：244 nm；柱温：30℃；进样量：20μl。结果：主成分与各杂质峰分离度良好。氯替泼诺浓度在0.001~1.02 mg·ml-1(r=0.9999)范围内与峰面积线性关系良好,平均回收率为99.9%,RSD为0.9%(n=9)。结论：该方法灵敏准确,色谱峰分离完全,可用于氯替泼诺妥布霉素混悬滴眼液的质量控制。%Objective:To develop an HPLC with gradient elution method for the determination of loteprednol etabonate and the re-lated substances in loteprednol etabonate and tobramycin compound ophthalmic suspension. Methods:HPLC with gradient elution was performed with Inertsil ph phenyl column (250 mm × 4. 6 mm, 5 μm). The mobile phase A and B was 0. 25% acetic acid solution-acetonitrile (80 ∶20) and acetonitrile, respectively. The flow rate was 2. 0 ml·min-1 and the detection wavelength was 244 nm. The column temperature was 30 ℃ and the injection volume was 20 μl. Results:The main ingredient and the related substances could be well separated. Loteprednol etabonate had a good linear relationship within the range of 0. 001-1. 02 mg·ml-1(r=0. 999 9), and the average recovery was 99. 9%(RSD=0. 9, n=9). Conclusion:The assay method is sensitive, accurate and convenient with good res-olution. It can be applied to control the quality of loteprednol etabonate and tobramycin compound ophthalmic suspension.