UPLC method for mycophenolate in injection

What is it about?

This study explores the ultra performance liquid chromatography for the quantification of mycophenolate mofetil (an immunosuppressant agent) in an injection formulation. Mycophenolate mofetil was separated from its seven specified impurities on HSS (high strength silica) C18 column (100 × 2.1 mm, 1.8 μm). The optimized mobile phase is compatible to mass spectrometry and hence the method can be equally run on LC-MS system for mass determinations. Efficient UV detection at 250 nm enabled determination of mycophenolate mofetil with no interference from placebo solution, diluent and other specified and unspecified impurities. The retention time of mycophenolate mofetil in the method was 1.79 min and all other impurities were eluted within in 8 min. The method optimization studies show that the selectivity of mycophenolate mofetil and its impurities was dependent on the type of organic solvent and its elution strength. The method was validated for specificity, linearity, precision, accuracy and robustness in range of 50 to 150 % of target analyte concentration (i.e. 62.50-195.31 μg mL-1). The linearity of peak area responses versus concentrations was demonstrated by linear least square regression analysis. An excellent linear relation exists between the concentration and detector response (R2 > 0.9999). Precision of the test method was proved with a relative standard deviation less than 1.0 %

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Why is it important?

A rapid, simple, sensitive, accurate and reliable RP-UPLC method was developed and validated for the determination of mycophenolate in injection formulations as per the ICH guidelines. In this method mycophenolate mofetil was separated from its seven impurities in 9 min of chromatographic run time and quantified with high degree of accuracy and precision. The retention time for mycophenolate was found to be 1.7 ± 0.2 min. Another advantage with this method is, it can be employed for mass determinations on LC-MS system, as its optimized buffer and mobile phase are compatible to mass spectrometry. The stability indicating power of the method was established through stress studies. All the degradation products formed during stress studies were well separated from the analyte peak which is evident from the peak purity data. The method discusses the nature of the molecule under stress conditions where it was shown that the compound is sensitive towards base hydrolysis (68.1 %), followed by oxidation (10 %), acid hydrolysis (3.7 %) and thermal degradations (2 %). The validation data meets the acceptance criteria for the validation parameters as per the current ICH (Q2R) and AOAC guidelines

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