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HPLC Method Validation For Pharmaceuticals: A Quick Review
The validation of HPLC methods is necessary to determine the suitability of the procedure for the intended use. The high-performance liquid chromatography or HPLC analysis is a chromatographic procedure which utilizes small particle columns to pump mobile phases at high pressure. The separation of the corresponding component is dependent on the interaction extent between the stationary phase and the solute component. The stationary phase components that have low affinity elute first.
The HPLC method development is a crucial aspect of the process of drug development due to its capabilities of providing better accuracy, sensitivity, resolution, recovery, preciseness, and reproducibility.
The article describes the method validation process of HPLC analysis. Read the following section for stepwise HPLC method validation process.
HPLC Method Validation
The effective method validation for HPLC analysis assists with quality control, regulatory control, and quality assurance of the drug development.
The validation protocol for HPLC method development is a standard document which indicates the validation procedure and approach. It ensures efficiency and consistency of the validation processes throughout the drug discovery and development. The validation protocol includes:
• Validation policy.
• Validation activities.
• Process documentation.
• Process characteristics.
• Acceptance criteria of processes.
• Relevant SOPs.
• Description of equipment and resources.
• Variation parameters.
Revalidation of the HPLC method development is achieved when a few method parameters fall out of the range. Additionally, any impurity, not previously known, can decrease the specificity of the method. The modifications in chemical quantity or equipment can also critically impact the validation. Hence, revalidation becomes necessary in these cases.
As per regulatory needs, various validation parameters are evaluated during HPLC analysis. These parameters are discussed below:
The specificity of the HPLC method quantitates the analytes when interference is present in the form of degradants, impurities, excipients, etc.
While the selectivity and specificity are interchangeably used, these two aspects are different. The selectivity is assessed for various chemical entities which may or may not be different from each other. The specificity is for one analyte.
Linearity checks the capability of a method to attain outcome which is directly proportional to the given range of HPLC sample concentration. A linear relationship in sample concentration and detector response is attained for assessment.
The accuracy of HPLC analysis is obtained as the closeness of accepted value (accepted reference or conventionally true) to the found value. It is not possible to assess the accurate actual or true value of particular HPLC sample preparation. But, the accepted value can be achieved by analyzing known concertation of the sample.
Precision is defined as the closeness in the measurements of various sampling obtained from the same homogenous HPLC sample under specified conditions. The precision is evaluated at three levels, repeatability, reproducibility, and intermediate precision.
• Repeatability is attained under the exact operating conditions of the same laboratory.
• Intermediate precision is obtained with different lab conditions such as different analysts, days, equipment, etc.
• Reproducibility is obtained by assessing the results of different laboratories.
The detection limit is assessed when the lowest analyte amount in a given HPLC sample can be distinguished but can’t be quantified in the exact value. There are various methods for assessing detection limits such as visual evaluation or noise ratio.
Quantitation limit is assessed when the lowest analyte amount in a given HPLC sample can be distinguished and quantified in exact value with accuracy and precision. It is can be assessed similar to detection limit such as through noise ratio.
The pharmaceutical discovery requires various HPLC method developments, which bring forth the need for method validations. These validations or HPLC analysis is necessary to ensure the analytical predictability and accuracy of drug products.
May 17, 2019