The characteristics of high-performance liquid chromatographic and gas chromatographic-electron capture assay systems are described for the separation of nitroglycerin from its metabolic and hydrolysis products: 1,2-dinitroglycerin, 1,3-dinitroglycerin, 1-mononitroglycerin and 2-mononitroglycerin. The methods quantitate the amount of parent compound and its major metabolites and were used to measure the rate of acid hydrolysis of nitroglycerin and its dinitro compounds. Assignment of the several peaks in the chromatograms is unequivocal as shown through the use of synthesized metabolites and hydrolysis studies.