Explained: HPLC Testing In Early Nonclinical Drug Development Workflows

Bioanalytical testing methods play a crucial role in supporting the needs of both nonclinical and clinical research studies. Bioanalytical services techniques are especially important during early nonclinical phases as the molecule of interest has to pass a stringent process to reach the subsequent non-clinical and clinical phases.

HPLC analysis is one such essential testing modality that has been employed over and over in the drug development process. HPLC stands for high-pressure liquid chromatography. From estimating dosage levels to identifying impurities in drug compounds, the HPLC method is widely used to verify a drug's safety and efficacy profile. From the initiation of an experiment to its completion, HPLC testing involves a lot of critical steps. Let us focus on the use of HPLC in early nonclinical drug development.

HPLC workflow during early nonclinical drug development

During the nonclinical phase, drugs are hardly studied in their pure form. Dissolved, diluted, or encapsulated drugs are among the most common modes of administration for an active pharmaceutical ingredient. During early nonclinical studies, drug molecules are evaluated for the estimation of the correct dosage levels. Such analysis involves the separation of the analyte from other impurities for the accurate quantification of the active ingredient. Such early nonclinical studies heavily rely on hplc lab.

All bioanalytical techniques consist of a separation method and a detector unit. The type of method depends on the chemical nature of the analyte and the mode of administration. HPLC is the most widely used separation method in the drug development process. HPLC is made up of a stationary phase, mobile phase, high-pressure pump, and detector unit. The stationary phase is a steel column comprising the absorbent material. A high-pressure pump drives the mobile phase, i.e., the solvent through the HPLC column. The analyte of interest is injected with the mobile phase and allowed to run through the HPLC column. Due to differences in affinity towards the HPLC column, each component of the drug compound travels at different speeds to move across the stationary phase.

The detector unit is placed at the end of the HPLC column. The choice of detector unit is based on the study sample to be analyzed. As each component of the drug molecule travels at its rates, the detector unit picks each of them accordingly. The detection data accumulated is then generated into graphical peaks through computer software. Each graphical peak corresponds to an individual component of the drug molecule, and the total curve area of distinct peaks corresponds to the concentration of that particular component. But it is critical to have prior knowledge of the retention times for accurate quantification of the drug components. Hence, HPLC method development and hplc method are vital before initiating any HPLC analysis.

Conclusion

HPLC testing has become a cornerstone technique in nonclinical and clinical research. The sensitivity and detection capabilities of HPLC testing are further increased with the addition of mass spectrometry as detector units, at times doubled with the addition of two mass spectrometry units, called LC-MS/MS. The advent of such innovative alternatives has made HPLC testing the go-to technique for early nonclinical drug development research.