Diatom cell quantity and their biochemical composition vary among species and are greatly affected by harvest stage or culture conditions. This study compares growth pattern, cell attachment, and biochemical composition of four diatoms suitable for abalone post-larvae: Navicula incerta, Proschkinia sp., Nitzschia sp., and Amphora sp. The four diatoms were grown in F/2 medium at 28.5 ± 1.4°C, under 62 ± 8 μmol photons m−2 s−1, at different original inoculating densities (0.05 × 106, 0.10 × 106, and 0.25 × 106 cells mL−1) and were harvested in log and stationary phase of growth for biochemical analysis. Total protein, carbohydrate, lipid, and ash composition, as well as fatty acid composition, were determined. All diatoms grew better when inoculated at 0.10 × 106 cell mL−1 with Proschkinia sp. reaching the highest cell density of 6.56 × 106 cells mL−1 in log phase. Amphora sp. had the highest cell attachment capacity when inoculated at 0.10 × 106 cell mL−1 (11,580 cells mm−2), whereas N. incerta had the lowest (7,750 cells mm−2). Protein and lipid (percent dry weight) contents were generally highest in cells during log phase of growth; Amphora sp. in log phase of growth had the highest lipid content of 9.74% DW, whereas significant differences in carbohydrate between the two growth phases were only observed for Proschkinia sp. Besides, all diatoms had higher energy contents in log phase of growth. There were no significant differences in ash content among the four diatoms. Polyunsaturated fatty acid (PUFA) content ranged from 23.25% to 38.62% of the total fatty acids, and the four diatoms tested were richer in n-3 PUFA than in n-6 PUFA. All the diatoms had significant quantities of 20:5n-3 (EPA) (between 12.69% and 17.68% of TFA), and Proschkinia sp., in log phase of growth, had the highest quantity of arachidonic acid (20:4n-6; ARA). The results highlight the influence of culture conditions and harvest protocols on diatom nutritive value and enabled a preliminary approach towards the selection of novel diatom species.