Adequate intake of n-3 fatty acids plays an important role in human health. The analysis of various blood lipids is used as a measure of fatty acid status in humans. Cheek cell phospholipids (PL) have also been proposed as biological markers, but are rarely used in clinical studies due to limitations in sample quality and quantity. An improved method for the analysis of cheek cell glycerophospholipid fatty acids is applied in a 29 d supplementation trial with 510 mg DHA daily. The DHA increases in cheek cell, plasma and erythrocyte glycerophospholipids are compared. High correlations are shown for glycerophospholipid DHA between cheek cells and plasma (r 0·88) and erythrocytes (r 0·76) before study commencement. After the daily supplementation of DHA, the half-maximal glycerophospholipid DHA level is reached after about 4 d in plasma, 6 d in erythrocytes and 10 d in cheek cells. The mean DHA increase (mol%) relative to baseline was most prominent in plasma (186 %), followed by cheek cells (180 %) and erythrocytes (130 %). Considering a lag phase of about 5 d, cheek cells reflect short-term changes in dietary fat uptake. Based on the data of the present study, they can be used alternatively to plasma and erythrocyte PL as non-invasive n-3 fatty acid status markers.