GC-MS and GC-MS/MS analytical methods were developed for the simultaneous detection of three 4(3H)-quinazolinone abused substances, including methaqualone, etaqualone, and 2-methoxyqualone, and applied to the analysis of suspected drugs and the hair of abusers, respectively. The standard solutions of methaqualone, etaqualone, and 2-methoxyqualone were analyzed by using the selected ion monitoring (SIM) and multiple reaction monitoring (MRM) modes, respectively, to determine the characteristic fragment ions and characteristic ion pairs of the three substances. The so established two methods were used to test the suspected drugs and hair samples in the actual cases. The established GC-MS analytical method detected components of methaqualone, etaqualone, and 2-methoxyqualone in the suspected drugs at levels of 11 μg/mg, 4.3 μg/mg, and 2.5 μg/mg, respectively. The established GC-MS/MS analytical method detected methaqualone, etaqualone, and 2-methoxyqualone in the hair at levels of 148 ng/mg, 565 pg/mg, and 58.2 ng/mg, respectively. The detection methods of GC-MS and GC-MS/MS established in this study are simple, rapid, and accurate, which can provide a reliable scientific basis for the detection of methaqualone, etaqualone, and 2-methoxyqualone in the suspected drugs and the hair of abusers.