Significance of HPLC Method Development, Validation, and Sample Analysis

Significance of HPLC Method Development, Validation, and Sample Analysis

High-performance liquid chromatography (HPLC) is the part of method development which may seem complex because of the high number of processes. protocols, parameters and equipment for successful completion of the process.

However, scientists working in laboratories where the processes are created will follow straightforward and planned steps to ensure the results are dependable and usable by the pharmaceutical industry. The HPLC method development, validation and sample analysis during drug development are necessary to expedite research and make a drug market ready with the required efficacy and safety profiles.

Let’s look at the steps that are typically adhered to by boutique contract research organizations: Selection of the HPLC Method

Researching previous literature to check method development and validation processes and successful results is the usually first part. Even the scientists who have previously performed the separation or have participated in similar research, need to consult their notes and checkpoints of the research. The reason is simple – it is necessary to increase the probability of the identified and selected HPLC system will help the scientists analyze the sample successfully.

Selection of the Initial Conditions The scientists use this step to determine the optimum conditions which are necessary to retain the analyte in the right concentration within the drug compound. The analyte factor should not be less than 0.5 or exceed 10-15. Negative factor leads to peak overlapping while excess retention can result in time-consuming analysis and poor detectability.

Optimization of the Selectivity During ​HPLC method development and validation, scientists consider the nature of the analytes with the aim to achieve adequate selectivity. They need to consider both mobile and stationary phases, which will further lead to reducing the trial chromatograms involved.

Optimization of the System Once scientists have achieved satisfactory selectivity, they follow it up with optimizing the system.

During optimization, the aim is to achieve the right balance between resolution and analysis time. Some of the parameters used are column-packing particle size, column dimensions, and flow rate.

The Right Method Validation

Correct method validation is an important step during the process as it ensures the safety profile and efficacy of a drug under observation. During the manufacturing process, it is the method validation that establishes the quality and sensitivity of a particular batch.

However, during HPLC analysis, not all the above-mentioned five steps are necessary. The factors which determine the steps are largely dependent on the nature of samples, analytes, previously established satisfactory separation and more.

Importance to HPLC Method Validation As has been established, method validation is necessary to establish the sensitivity of the selected drug profile. While segregating analysis of method validation from method development is important, it is equally important to establish the parameters for measurements and then follow the above steps to achieve accuracy and precision for all processes. Scientists can conduct method development and validation simultaneously, however, it is vital to remember, the two processes are different from each other and are followed to get the correct, viable and accurate results to aid in drug development.