After establishing liquid chromatography detection method and should be demonstrated, usually it requires a full demonstrate specificity, linearity, accuracy, precision, recovery, sensitivity of verification carried out by a simplified method. Before the final demonstration method, the written analysis plan should be prepared and checked to make it clear and rigorous. The actual argumentation procedure varies according to the method, and the time varies from one day to two weeks. Ideally, methodological evidence can identify potential problems that may arise from differences in instrumentation and operator.
Inter-column reproducibility may be one of the main problems in daily HPLC analysis. The same method may have different results on different batches of columns, and any unexpected results should be studied to find out the cause and avoid similar errors in future work, so establish a complete liquid chromatograph method and It is especially important to argue with it.
1. Data should be prepared to indicate the performance of the required method;
2. There should be written analytical steps for other operators;
3. Systematically demonstrate the performance of the method with more than one system or operator, including the desired component and the desired concentration of the analyte; compare experimental data between different time and different laboratories;
4. Data on the expected life of the column and the reproducibility between columns should be obtained;
5. Study abnormal results to correct potential problems;
6. Study all conditions that affect separation (temperature, mobile phase composition, pH, etc.);
Limit the limits of these conditions; propose possible measures for possible problems (insufficient separation of key peaks; increase in run-time, increase in peak retention, etc.).
The requirements of these conditions apply to rigorous HPLC standard methods that are accurate, accurate, reliable, and transferable. In other cases, it may be necessary to only require a successful separation or to respond quickly and “roughly†to a particular problem. In this case, there are many suggestions, and it is sufficient to understand and know the true goal of a method to establish a solution.
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