The so-called systematic toxicological analysis (STA) aiming at simultaneous analysis of as many toxicologically relevant compounds in biosamples as possible is an important part of routine analysis in clinical and forensic toxicology. Gas chromatography-mass spectrometry and liquid chromatography with diode array detection have been the most widely used techniques for this purpose. However, in recent years STA methods based on liquid chromatography coupled to mass spectrometry (LC-MS) or tandem mass spectrometry (LC-MS/MS) have become increasingly important, although their widespread use is still hampered by the lack of a universal reference library of mass spectra that can be used on all major instrument platforms. In this review, LC-MS(/MS) methods for STA in urine and/or blood published in the past 6 years will be compared and discussed with regard to sample preparation, separation, instrument types used for mass spectrometric detection, and method validation. In addition, different approaches to achieving the goal of a universal reference library will be summarized.