Specific dietary polyunsaturated fatty acids (PUFAs) and long chain PUFAs (LCPUFAs) elevate femur calcium content and enhance calcium balance. Mother's milk is associated with enhanced calcium balance and contains LCPUFAs; arachidonic acid (AA), and docosahexanoic acid (DHA). However, the effect of AA and DHA on calcium metabolism and other bone minerals during infancy is unknown. Thus, piglets received one of four formulas (15 d): control or with AA:DHA (0.5:0.1 g, 1.0:0.2 g, or 2.0:0.4 g/ 100 g of fat). Calcium absorption, femur mineral composition, and urinary mineral excretion. Main effects identified using two-way analysis of variance (ANOVA) and post hoc analysis conducted using Duncan's multiple range test. Significant effects of diet were observed in femur calcium and zinc content, but not calcium absorption, urinary mineral excretion, femur ash weight, femur phosphorus, or femur magnesium content. The piglets receiving AA:DHA as 1.0:0.2 g/100 g of fat had 1.9% higher mg calcium/g of ash, but 8.6% lower g zinc/g of ash compared with control. Thus, AA:DHA in a ratio of 5:1 does not affect mineral accretion, but AA plus DHA, in amounts similar to the upper limit of human milk, might be detrimental to bone mineralization over time due to lower zinc.