An in vitro gastrointestinal human digestion model, with and without additional rapeseed oil, was used to measure the bioaccessibility of the major lipophilic nutrients enriched in chloroplasts: β-carotene; lutein; α-tocopherol; and α-linolenic acid. Chloroplast-rich fraction (CRF) material for this work was prepared from post-harvest pea vine field residue (pea vine haulm, or PVH), an abundant source of freely available, underutilised green biomass. PVH was either steam sterilised (100 °C for 4 min) and then juiced (heat-treated PVH, or HPVH), or was juiced fresh and the juice heated (90 °C for 3 min) (heat-treated juice, or HJ); the CRF from all biomass treatments was recovered from the juice by centrifugation. The impact of postharvest heat treatment of the biomass (HPVH), or of heat treatment of the juice (HJ) derived from the biomass, on the retention and bioaccessibility of the target nutrients was determined. The results showed that both heat treatments increased the apparent retention of β-carotene, lutein, α-tocopherol, and α-linolenic acid in the CRF material during digestion. The presence of edible oil during digestion did not dramatically affect the retention of these nutrients, but it did increase the bioaccessibility of β-carotene, lutein, and α-tocopherol from CRF material derived from heated biomass or juice. The presence of oil also increased the bioaccessibility of β-carotene, but not of lutein, α-tocopherol, or α-linolenic acid, from fresh CRF material.