Guayule, a shrub native to the Chihuahuan desert, is a natural source of high quality, hypoallergenic rubber. Unlike rubber trees that produce rubber in laticifers, the rubber in guayule is produced in parenchyma cells of the bark tissue of stems and roots. Consequently, guayule tissue must be mechanically broken before the rubber can be extracted and analyzed. Since rubber extraction and analysis is time-consuming, progress towards increasing the rubber content of guayule through breeding or better cultivation practices has been limited by the slow rate of sample processing. To address the need for faster and more efficient sample throughput, conditions were optimized for automated extraction of dried guayule tissue using accelerated solvent extraction (ASE) and rapid methods were developed to replace gravimetric determination of resin and rubber content. For resin analysis, ultraviolet absorbance was used to determine resin concentration after ASE of the tissue with acetone or acetonitrile. For rubber analysis, evaporative light scattering (ELS) was used to determine the amount of rubber recovered after ASE of the tissue with cyclohexane. Extraction of guayule tissue with high latex rubber content verified that the amounts of resin and rubber determined by these methods were similar to the amounts determined gravimetrically. Since these methods automate extraction and increase the speed of resin and rubber quantification, they could be used in combination with ASE to increase the throughput and efficiency of guayule evaluation in germplasm enhancement and agronomic improvement programs.