Figure 5.
SEC-MALLS analyses of choroidal HA. (A) Chromatograms from SEC-MALLS of choroid samples from day 8 recovering and contralateral control chick eyes. Lines: refractive index traces indicating the amount of HA. Individual points: Mw values calculated from the laser light scattering data for each time (eluate) interval. Each sample represents four choroids pooled separately from control (thin line, open circles) and recovering eyes (thick line, solid circles). (B) To confirm that the SEC-MALLS signals detected were caused by HA, a portion of each sample was treated at 37°C for 24 hours with 6 U of S. hyalurolyticus hyaluronidase, which specifically digests only HA. Refractive index (concentration) traces are shown for a choroid sample from a day 7 recovering eye that was untreated (thick line) or hyaluronidase-treated (thin line). Signal reduction in this digested sample was 96%. Overall, the HA content of samples was at least 91% ± 5% (n = 28).