Ceramides (Cer) are implicated in obesity-associated skeletal muscle and perhaps adipocyte insulin resistance. We examined whether the sphingolipid content of human subcutaneous adipose tissue and plasma varies by obesity and sex as well as the relationship between ceramide content and metabolic indices. Abdominal subcutaneous adipose biopsies were performed on 12 lean adults (males = 6), 12 obese adults (males = 6) for measurement of sphingolipid content and activity of the main ceramide metabolism enzymes. Blood was sampled for glucose, insulin (to calculate homeostasis model assessment-estimated insulin resistance (HOMAIR)) adiponectin, and interleukin-6 (IL-6) concentrations. Compared to lean controls, total ceramide content (pg/adipocyte) was increased by 31% (P < 0.05) and 34% (P < 0.05) in obese females and males, respectively. In adipocytes from obese adults sphingosine, sphinganine, sphingosine-1-phosphate, C14-Cer, C16-Cer, and C24-Cer were all increased. C18:1-Cer was increased in obese males and C24:1-Cer in obese females. For women only, there was a negative correlation between C16-Cer ceramide and plasma adiponectin (r = −0.77, P = 0.003) and a positive correlation between total ceramide content and HOMAIR (r = 0.74, P = 0.006). For men only there were significant (at least P < 0.05), positive correlations between adipocyte Cer-containing saturated fatty acid and plasma IL-6 concentration. We conclude that the sexual dimorphism in adipose tissue behavior in humans extends to adipose tissue sphingolipid content its association with adiponectin, IL-6 and insulin resistance.