Vitellogenin (Vg) concentration was determined in pupal hemolymph of female southwestern corn borer, Diatraea grandiosella (Dyar). Vitellogenin concentration increased significantly with pupal age. Vitellogenin level reached 7 mg/ml on day 7 after pupation. The vitellogenin was separable into 2 apoproteins on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), with molecular weights of 145,000 for apoVg-I and 38,000 for apoVg-II. SDS-PAGE analysis revealed that vitellogenin polypetides in female pupal hemolymph appeared to be identical to those from eggs, but were absent in male and early female pupal hemolymph. A method to quantify vitellogenin by indirect ELISA is described. Results from this study and those of a previous study suggest that juvenile hormone does not have a role in vitellogenesis but does so in choriogenesis.