Lipids, a class of biomolecules, play a signi cant role in the physiological system. In this study, gas-phase hydroxyl radicals (OH•) and atomic oxygens (O) were introduced into the collision cell of a triple quadruple mass spectrometer (TQ-MS) to determine the positions of the double bond in unsaturated phospholipids. A microwave-driven compact plasma generator was used as the OH•/O source. e reaction between OH•/O and the precursor ions passing through the collision cell generates product ions that correspond to the double bond positions in the fatty acyl chain. is double bond position speci c fragmentation process initiated by the attachment of OH•/O to the double bond of a fatty acyl chain is a characteristic of oxygen attachment dissociation (OAD). A TQ-MS incorporating OAD, in combination with liquid chromatography, permitted a high throughput analysis of the double bond positions in complex biomolecules. It is important to know the precise position of double bonds in lipids, since these molecules can have widely different functionalities based on the position of the double bonds. e assignment of double bond positions in a mixture of eight standard samples of phosphatidylcholines (phospholipids with choline head groups) with multiple saturated fatty acyl chains attached was successfully demonstrated.