Artificial neural network (ANN) analysis is a new technique in NMR spectroscopy. It is very often considered only as an efficient ‘black-box’ tool for data classification, but we emphasize here that ANN analysis is also powerful for data quantification. The possibility of finding out the biochemical rationale controlling the ANN outputs is presented and discussed. Furthermore, the characteristics of ANN analysis, as applied to plasma lipoprotein lipid quantification, are compared to those of sophisticated lineshape fitting (LF) analysis. The performance of LF in this particular application is shown to be less satisfactory when compared to neural networks. The lipoprotein lipid quantification represents a regular clinical need and serves as a good example of an NMR spectroscopic case of extreme signal overlap. The ANN analysis enables quantification of lipids in very low, intermediate, low and high density lipoprotein (VLDL, IDL, LDL and HDL, respectively) fractions directly from a 1H NMR spectrum of a plasma sample in <1 h. The ANN extension presented is believed to increase the value of the 1H NMR based lipoprotein quantification to the point that it could be the method of choice in some advanced research settings. Furthermore, the excellent quantification performance of the ANN analysis, demonstrated in this study, serves as an indication of the broad potential of neural networks in biomedical NMR.