In recent years, there has been a stark decline in bee colony populations. Many studies believe neonicotinoids to be a significant contributor to this problem. Neonicotinoids are insecticides that are often used in farming or agricultural settings. One of the neonicotinoids suspected to play a role in the bee colony decline is Imidacloprid. Imidacloprid is believed to directly negatively affect the navigational skills of bees. It interrupts their homing abilities, making it difficult for them to relocate their hive after coming in contact with Imidacloprid. Previous research projects have been conducted to study the affect of Imidacloprid on bees, but the proportion of Imidacloprid to the bee itself is not realistic of what they would be affected by in a veritable agricultural setting. Here we demonstrate an extraction protocol based on sample clean-up with both ion-exchange and reversed phase separations prior to targeted gas chromatography-mass spectrometry (GC-MS) analysis. This protocol is similar to the QuEChERS (Sigma, St. Louis, IL) method, however, the extraction protocol presented here shows how capturing and concentrating the Imidacloprid while washing away the other matrix components can help improve sensitivity and selectivity for Imidacloprid. Our targeted detection strategy involves selected ion monitoring (SIM) to decrease limits of detection. Initial findings show that C-18 separation is optimized by adding a centrifugation step prior to SPE (solid phase extraction). As we continue to develop a targeted detection strategy for Imidacloprid, the hope is that the strategy will be used in future farming and agricultural environments to determine if the level of neonicotinoids being used in specific fields is detrimental to the bee colony population in the area.