Comprehensive investigation of matrix effect evaluation approaches for reliable quantification in bioanalysis using...

Area of Science:

• Analytical Chemistry
• Mass Spectrometry
• Chromatography

Background:

• Matrix effect (ME) evaluation is essential for ultra-high performance liquid chromatography coupled with mass spectrometry (UHPLC-MS) method validation.
• Rotational (analyte-dependent) and translational (analyte-independent) MEs impact calibration curves.
• Common ME evaluation strategies include post-extraction addition and calibration curve slope comparison.

Purpose of the Study:

• To systematically assess the suitability of the calibration curve slope approach for quantifying ME.
• To compare the calibration curve slope approach with the post-extraction addition method.
• To develop a novel approach for ME calculation that includes translational effects.

Main Methods:

• Systematic comparison of two ME evaluation strategies using ESI-UHPLC-MS/MS in positive and negative ionization modes.
• Analysis of 26 compounds in serum.
• Assessment of various calibration models (1/X^0, 1/X, 1/X^2 weighting, logarithmic transformation).

Main Results:

• The calibration curve slope approach alone can underestimate ME compared to the post-extraction addition method.
• A new ME calculation approach was developed, incorporating translational ME from the calibration curve intercept.
• The novel approach demonstrated accuracy across three matrices and two instrumental platforms.

Conclusions:

• Relying solely on the calibration curve slope approach for ME estimation is inadvisable.
• A novel intercept-based equation accurately calculates translational ME.
• The total ME, combining slope and intercept contributions, closely matches results from the post-extraction addition method.