When conducting biomedical research, biological samples have the characteristics of less sampling, low drug concentration, endogenous substances, and individual differences. These factors may affect the determination of biological samples. Therefore, it is necessary to establish appropriate biological sample analysis methods. The verification of these biological analysis methods and the analysis of test samples should comply with the GLP principle, so GLP biological analysis is very important. After the analytical method is established, method verification must be carried out. The purpose of verification is to ensure that the analytical method is suitable for relevant sample analysis.
The methodological verification of biological sample analysis is mainly divided into three types: comprehensive verification, partial verification and cross-validation. Today we will briefly understand these three verification methods.
Macromolecular biopharmaceuticals are derived from cell culture, and their potency and immunoreactivity vary greatly from batch to batch. In most cases, there is no real “pure” macromolecular drug reference substance. This requires the preparation of standard samples and quality control samples in the same batch as the pre-clinical and clinically administered control substances. When changing batches, analysis and identification and bioanalytical evaluation of the reference substance must be performed before use to confirm that the analysis method has not changed. For an analytical method that is essentially a new drug, and the analytical method established for the first time, a comprehensive analytical method verification is required. When a metabolite is added to the analytical method for quantitative analysis, a comprehensive analytical method verification should also be considered.
The establishment and verification of bioanalytical methods are essential to obtain reliable experimental results. When conducting GLP biological analysis, the personnel participating in the analysis of biological samples under GLP conditions include operators and checkers, and must be trained in GLP regulations and related skills and passed the assessment. It is necessary to understand the relevant rules and regulations before the test. The analytical instruments used, such as mass spectrometry, liquid phase, electronic balance, precision pipette, etc., must also be within the validity period of verification or forced inspection. Medicilon’s biological analysis service department has a professional scientific research team, the analysis laboratory is equipped with advanced equipment, implements comprehensive information management, experimental research meets the requirements of FDA/CFDA GLP standards, and can provide customers with small molecule drugs, biological agents, Screening and development of vaccines and biomarkers, as well as preclinical and clinical research.
Methodological verification is the basic guarantee for the reliability of the entire experimental data, so it is the key content of each laboratory’s standard operating procedures (SOP) . When the verified analysis method is changed, the verification of part of the analysis method can be considered, such as the transfer of biological sample analysis method between laboratories, the change of quantitative concentration range, the change of biological medium, the rare biological medium, and the verification of the analysis method after compound administration Specificity. At this time, all changes should be reported, and the reasons for the scope of re-verification or partial verification should be explained.
Compare the validation parameters of the two analytical methods. It is often used for the change of different analysis methods for the same substance, such as the change from LC/MS to ELISA, or when two or more laboratories analyze the same substance according to the same protocol. For data obtained by using different analytical methods in the same laboratory or using the same analytical method in different laboratories, it is necessary to compare the data and cross-validate the analytical methods. As far as possible, cross-validation should be carried out before the analysis of the research samples. Two analytical methods should be used to analyze the same quality control sample or real sample. For quality control samples, the accuracy of the two analytical methods should be within 15%; for research samples, the accuracy of at least 67% of the samples should be within 20%.
The establishment of a reliable and reproducible quantitative analysis method is one of the keys to biomedical research. In order to ensure the reliability of the analysis method, method verification must be carried out. The establishment, verification and application of biological sample analysis methods are an important part of preclinical pharmacokinetics and phased clinical trial research, which is related to the scientificity and correctness of follow-up research, and investigations should be started during the establishment of analytical methods. Confirmation at the verification stage, and monitoring sample analysis stage.