AbstractThe in vitro method in use for the determination of β‐carotene bioaccessibility involves simulated gastrointestinal digestion followed by ultracentrifugation to separate the micellar fraction containing bioaccessible β‐carotene and its quantitation. In this study, the suitability of two alternatives viz., membrane filtration and equilibrium dialysis were examined to separate the micellar fraction. Values of β‐carotene bioaccessibility obtained with the membrane filtration method were similar to those obtained by the ultracentrifugation method. Equilibrium dialysis was found not suitable for this purpose. Among the vegetables analyzed, fenugreek leaves had the highest content of β‐carotene (9.15 mg/100 g), followed by amaranth (8.17 mg/100 g), carrot (8.14 mg/100 g) and pumpkin (1.90 mg/100 g). Percent bioaccessibility of β‐carotene ranged from 6.7 in fenugreek leaves to 20.3 in carrot. Heat treatment of these vegetables by pressure cooking and stir‐frying had a beneficial influence on the bioaccessibility of β‐carotene from these vegetables. The increase in the percent bioaccessibility of β‐carotene as a result of pressure‐cooking was 100, 48 and 19% for fenugreek leaves, amaranth and carrot, respectively. Stir‐frying in presence of a small quantity of oil led to an enormous increase in the bioaccessibility of β‐carotene from these vegetables, the increase being 263% (fenugreek leaves), 192% (amaranth leaves), 63% (carrot) and 53% (pumpkin).