1. In platelet rich plasma (PRP), chondroitin 4-sulfate release from platelets occurred after stimulation with ADP (5 microM), collagen (5-10 micrograms/ml), or adrenaline (10 microM). Release started within 60 s and maximum release (0.7-2.0 mg/l) was reached within 180 s. TXA2 formation and dense granule release reached a maximum within 90 s after stimulation. 2. Using washed platelets (1.5 x 10(8) cells/ml), the platelet responses were faster. Release of chondroitin 4-sulfate and TXA2 started within 20-30 s after thrombin addition (100 mU/ml). Maximum release was reached within 60 s in both cases. Dense granule release started in the first 5 s of stimulation (34.6 +/- 12.4%) reaching maximum secretion (74.4 +/- 8.7%) within 60 s. 3. Our results demonstrate that maximal chondroitin 4-sulfate release occurs after the dense granule release reaction in both PRP and washed platelets. This observation suggests that chondroitin 4-sulfate is unlikely to be stored in the dense granules but may be stored in the alpha-granules.