There is evidence that carotenoids, the fat-soluble pigments in fruits and vegetables, are linked to improvements in certain chronic diseases including agerelated macular degeneration, cell-transformation and cardiovascular diseases. It is also known that for humans the bioavailability of carotenoids present in fruits and vegetables is comparatively low and strongly dependent on carotenoid species, chemical structure and food matrix. In this study, an in vitro digestion approach was used to screen the bioaccessible carotenoid content of raw and processed vegetables after three main digestion steps with saliva, gastric- and duodenal juices. The experimental setup was based on the in vitro digestion model of Oomen et al. (2003). First raw or cooked vegetable samples were chopped and incubated at 37°C for 10 min in a shaking water bath with human saliva (pH 6.5; α-amylase). Afterwards, simulation of the gastric phase was achieved by addition of gastric juice (pH 1.0; porcine pepsin) and incubated for an additional 1 h. In a third step, duodenal juice (pH 7.8; porcine pancreatin, porcine lipase) and porcine bile solution (pH 8.0) were added. Samples were then incubated under nitrogen for 2 h, centrifuged and the supernatants were freeze-dried. The six most important nutritional carotenoids (lutein, zeaxanthin, β-crypthoxanthin, α-carotene, β-carotene, lycopene) were quantified by HPLC-UV/VIS. The carotenoids were separated in 47 min on a C30 column with a methanol/MTBE gradient and detected by diode array detection (DAD). Freeze-dried supernatants were redissolved in the eluent, centrifuged and filtered before injection. External standards were used for quantification. In raw green leafy vegetables the carotenoid accessibility was low compared to orange/red colored vegetables and ranged between 4-16% for lutein, 7-25% for zeaxanthin and 12-22% for β-carotene. The bioaccessibility of carotenes and xanthophylls from raw red and orange vegetables was considerably higher and up to 57%. Generally, an increased carotenoid accessibility was observed after thermal processing. This adapted in vitro digestion method is a promising tool to investigate the carotenoid bioaccessibility from vegetables.