Abstract: Described herein are automated and customizable sample preparation and analysis systems for detection and quantification of biomarkers (e.g., metabolites and/or lipids) in biological samples (e.g., blood, serum, or plasma) in a clinical setting. The automated systems are controlled by scripts that integrate communication between the components of the sample preparation system. Also described herein are mass spectrometry-based analytical methods featuring efficient system calibration and sample analysis that provide for accurate quantification of a set of markers in biological samples. The methods are capable of automatic high sample throughput in a clinical setting for detection and quantification using a mass spectrometry system and high performance liquid chromatography column.