Although orange juice is a popular beverage worldwide, fruit distribution, storage, and processing can facilitate fungal infection by Penicillium digitatum; leading to the production of tremorgenic alkaloids, specifically tryptoquialanines A (TA) and B (TB). An Analytical method was developed and validated based on QuEChERS and LC-MS/MS analysis to determine the levels of TA and TB in fresh, industrial, and homemade orange juices. Excellent linearity was observed in the method over a high range of 1-1000 mug/kg and low range of 1-75 mug/kg with R(2) >/= 0.998. The LOD and LOQ were 1 and 3 mug/kg, respectively. Recoveries showed values between 57 and 83 %, with RSD </= 13 %. Our data indicated a higher prevalence of mycotoxin TA in fresh and industrial orange juices. Reduction in TA and TB content after thermal and HPP treatments were </= 32 %. However, thermal treatment was more effective in reducing TA and TB contents. CI - Copyright (c) 2023 Elsevier Ltd. All rights reserved.