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Bioanalytical Method Development: High-Quality Data for your IND or NDA Filing to US FDA

Analytical method development is a procedure for reliable detection of analytes of interest. High-grade method development ensures that the method is suitable for full validation and high-quality analysis of drugs, metabolites, and biomarkers in biological matrices.As the number of drugs released in the market is increasing by the day, so is the need for method development and validation. These new drugs may be compounds never used before or modification of existing drugs. Inability to relate analytical procedures, as well as reference and internal standards for new drugs, hinders their thorough R&D. Hence, it becomes necessary to develop new analytical methods for studying the effect of drug compounds. These methods are designed to understand the identity, characteristics, purity, and potency of drug compounds.

During drug research, the developed analytical methods are utilized to measure and define the required dosage, acceptable impurity, observed stability, and bioavailability of the drug. Overall, analytical method development ensures product consistency, allowing the release of reliable drug products in the market.

Bioanalytical Method Development and Validation: From Drug Discovery to Clinical Trials

Drug development is an intricate process involving several phases. Analytical method development starts closer to the beginning of the drug development process at the discovery stage itself. Method development for a New Chemical Entity (NCE) enables analysis in biological matrices for evaluation of drug pharmacology properties, such as bioavailability.  Generally, a non-validated method is deployed at this stage of early drug discovery. As the drug progresses to IND enabling studies, however, further method development is required given the need for method robustness and full validation. Even more method development may be needed eventually, due to the change in matrices from animal species to human, as the drug moves farther into Phase I.

Drug Discovery

During drug discovery, scientists identify novel treatment targets, synthesize new molecules, screen these molecules in ‘in vivo’ and ‘in vitro’ assay, and study physicochemical aspects of the molecules. Multiple NCE are tested to determine the compound that may be best suited for the development of a drug, and each of these NCE requires prompt method development.

Preclinical

Several analytical methods are developed for testing the samples of different compounds in a variety of species. These methods eventually would be adapted to human samples. Adequate method development during the preclinical stage is essential for a straightforward transition from animal species to Clinical phase.

Clinical

During clinical trials, method development helps mark a safe dose for human consumption. These trials are performed on human volunteers to ensure the stability, reliability, and effectiveness of the drug products and associated compounds. Usually, methods developed during the preclinical phase are modified and applied to human plasma samples at this stage. Further analytical method development is often required as more is discovered about the drug and its metabolism in this phase, necessitating the analysis for metabolites as well.

Bioanalytical Method Development and Validation by NorthEast BioLab

Various method development techniques are utilized to analyze drugs and metabolites in biological fluids with high precision and accuracy. Other parameters such as analyte stability, interaction with additional medications are also determined. Selecting the right analytical method is imperative for the sample analysis data integrity and drug advancement in the right direction.

LC MS MS

LC MS MS has become one of the most popular drug analysis technique given rapid method development on the platform. Liquid Chromatography with tandem mass spectrometry relies on their combined strengths. While liquid chromatography has excellent separation capabilities, triple quadrupole mass spectrometry offers nuanced selectivity and sensitivity.

HPLC/UV

These methods use a UV detector to analyze samples and measure their absorption ability for analyte identification. The ultraviolet (UV) detector measures the absorbance of monochromatic light of a fixed wavelength in the UV or visible wavelength range against a reference beam and relates the magnitude of the absorbance to the concentration of an analyte in the eluent passing through a flow cell contained within the instrument. The HPLC UV method is preferable for solutions that have an unknown molecule count. Similarly, drugs with analytes suitable for UV detection have good chromophore due to unsaturated bonds, aromatic groups, or functional groups containing heteroatoms.  Although in some instances, this technique may be necessary, analysts prefer LC MS method development over UV, as detection can be time-consuming and transition to Clinical samples difficult due to inconsistent interferences in plasma from subject to subject.

HPLC/FL

HPLC/FL method development can be extremely swift due to the selective nature of this technique. Here, it’s possible to detect even a single analyte molecule in the flow cell. Put differently, fluorescence sensitivity can be 100x more than that of the UV detector for some specific compounds. This is typically an advantage in the measurement of fluorescent species in samples.

ELISA

Enzyme-linked immunosorbent assay is a method utilized for specific proteins. The technique captures an antigen on a solid surface which is then complexed to an antibody. The detection is achieved by assessing the activity of the enzyme.  Although there has been a notable shift towards using LC/MS/MS analysis for large molecules, it is still quite challenging to achieve the same sensitivity levels as with ELISA.  That said, method development with ELISA can be tricky as other endogenous substances in the biological matrices interfere with analyte detection.

When it comes to analytical method development, scientists have several critical choices to make from a variety of sample preparation techniques, chromatographic conditions, and detection systems. We make every effort to develop a robust and straightforward method that can be used frequently for producing reliable results. The first step in method development is to select a suitable sample clean-up procedure such as simple protein precipitation. Other available options include liquid extraction or solid phase extraction. In some case, it may be necessary to use an even more involved procedure such as chemical derivatization. Once a proper sample clean-up technique is decided, suitable chromatographic conditions are selected next, followed by the determination of an appropriate detection system. Although older detection systems such as ultraviolet (UV) and fluorescence are still in use, mass spectrometry has emerged as the detection system of choice over the last couple of decades due to its high selectivity.

Basis of Bioanalysis

The Basics of Bioanalysis

Bioanalysis is an essential tool in drug discovery and development for determining the concentration of drugs and their metabolites as well as various pharmacodynamics biomarkers in biological fluids. In these analyses, scientists use developed an….

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Sturdy Bioanalytical Method Development and Validation for Independent Data Reproducibility

Method development is not a cut-and-dry science, and rather requires some art that only comes with experience. Our scientists have 15+ years of experience in analytical method development for drugs, metabolites, and biomarkers. We emphasize robust and straightforward method development and validation to ensure that multiple analysts and laboratories can reproduce the method independently under minimal supervision. For instance, we always start with a thorough sample clean up, and clear chromatographic separation as these two steps are critical for selectivity and reproducibility of LC-MS methods. Similarly, we focus on careful selection of mass spectrometric parameters essential for method sensitivity. Our veteran team’s fundamentals focused approach eases the progression of these methods from preclinical to clinical stage with minimal changes, leading to streamlined clinical sample analysis and accelerated data submission.

NorthEast BioLab is a drug testing laboratory that helps clients follow a scientific and known approach for various analysis. Our team believes in finding proper methods for superior outcomes and reliable drug testing. Expertise in techniques such as LC/MS/MS, HPLC, and ELISA offers us the capability of delivering solid, quantifiable results. In addition to our experience and efficiency, we provide complete transparency to ensure the reliability of critical data and reduce the turnaround time of method development and validation.

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