Extensive assignments of resonances in the 600 MHz 1H-NMR spectra of cerebrospinal fluid are reported. These have been achieved by the measurement of a combination of two-dimensional experiments comprising homonuclear J-resolved, COSY45, and double-quantum filtered COSY (DQCOSY) spectra. By these means the previous total of 18 endogenous metabolites, of which in general only selected resonances have been assigned, has been augmented to 46 molecules including all of the resonances of both alpha- and beta-anomers of glucose. With only a few exceptions all resonances have been assigned for all of the metabolites. In addition, the effect of freeze-drying on the 600 MHz 1H-NMR spectrum of human cerebrospinal fluid (CSF) is presented using both lyophilization with reconstitution into either H2O or D2O. Freeze-drying and reconstitution into H2O causes a significant sharpening of many small molecule resonances, including notably those of glutamate and glutamine as well as other amino acids and in addition causes the loss of volatile components, principally acetone. Further exchange of the H2O solvent by D2O causes no additional changes in the spectra.