Malnutrition, as a result of deficiency in essential nutrients in cereal food products and consumption of a poorly balanced diet, is a major challenge facing millions of people in developing countries. However, developing maize inbred lines that are high yielding with enhanced nutritional traits for hybrid development remains a challenge. This study evaluated 40 inbred lines: 26 quality protein maize (QPM) lines, nine non-QPM lines, and five checks (three QPM lines and two non-QPM lines) in four optimum environments in Zimbabwe and South Africa. The objective of the study was to identify good-quality QPM inbred lines for future hybrid breeding efforts in order to increase the nutritional value of maize. The QPM lines had a lower protein content (7% lower) than that of the non-QPM lines but had 1.9 times more tryptophan and double the quality index. The lysine- and tryptophan-poor α-zein protein fraction was 41% lower in QPM than in non-QPM, with a subsequent increase in γ-zein. There was significant variation within the QPM inbred lines for all measured quality characteristics, indicating that the best lines can be selected from this material without a yield penalty. QPM lines that had both high protein and tryptophan levels, which can be used as parents for highly nutritious hybrids, were identified.