A basic nonapeptide P2 (antiflammin-2, HDMNKVLDL) which is identical to a portion of the amino acid sequence (residues 246–254) of lipocortin I, has been described to have antiinflammatory activity in a rat paw edema model (Nature 335: 726–730 [1988]). P2 (0.05 μM) was also reported to inhibit porcine pancreatic phospholipase A₂ (PLA₂). The effect of synthetic P2 (98% pure) on PLA₂ was evaluated in two assay systems. Using porcine pancreatic PLA₂ and phosphatidylcholine/deoxycholate mixed micellar substrate, P2 (0.005–50 μM) had no effect on PLA₂ activity, even in the presence of 2-mercaptoethanol to prevent peptide oxidation. In another assay, using human synovial fluid PLA₂ as the enzyme and [¹⁴C]-oleate-labelledE. coli substrate, P2 (0.005–50 μM) had no significant effect on PLA₂ activity. A reported PLA₂ inhibitor, manoalide, was a potent inhibitor of PLA₂ in both assay systems. On the basis of these results, we conclude that P2 is devoid of PLA₂ inhibitory activity.