Improving LC/MS/MS-based bioanalytical method performance and sensitivity via a hybrid surface barrier to mitigate...

Area of Science:

• Analytical Chemistry
• Pharmacokinetics
• Bioanalysis

Background:

• Accurate pharmacokinetic (PK) determination is crucial for drug discovery and development.
• Liquid chromatography-mass spectrometry (LC/MS) is the standard for PK studies due to its sensitivity and selectivity.
• Interactions between analytes and metal components in LC systems can compromise bioanalysis sensitivity and peak shape.

Purpose of the Study:

• To evaluate the benefits of an inert hybrid surface technology (HST) in mitigating unwanted analyte-metal interactions in LC systems.
• To assess the impact of HST on the bioanalysis of steroid phosphate drugs and an EGFR inhibitor in human plasma.
• To compare the performance of an HST-equipped LC system against a conventional LC system.

Main Methods:

• Employed an inert hybrid surface technology (HST) on metallic components within the LC flow path, column frits, and column wall.
• Conducted comparative bioanalysis of dexamethasone phosphate, hydrocortisone phosphate, and gefitinib in human plasma using both HST-equipped and conventional LC systems.
• Evaluated peak shape (width, tailing factor), assay sensitivity, limit of detection, and reproducibility (coefficient of variation).

Main Results:

• For steroid phosphates, HST reduced peak width by 20% and peak tailing factors by up to 30%.
• Assay sensitivity improved significantly (7.5- and 10-fold) for steroid phosphates, enhancing the limit of detection.
• For gefitinib, HST reduced coefficients of variation (%CV) from >10% to <5% at the lower limit of quantification, improving reproducibility.

Conclusions:

• Inert hybrid surface technology effectively mitigates adverse analyte-metal interactions in LC systems.
• HST significantly improves bioanalytical method performance, including peak shape, sensitivity, and reproducibility.
• This technology enhances the accurate determination of drug pharmacokinetics, particularly for low-concentration analytes in biological matrices.