Figure 3:Mild acid hydrolysis-fluorometric anion-exchange chromatography analysis (MH-FAEC). The PEB and PAB crude samples (1 mg as BSA) were mild acid hydrolyzed and simultaneously labeled with DMB. Released and labeled oligo/polySias were separated with DNA-Pac PA-100 anion exchange chromatography and eluted with a gradient of NaCl (0-15 min, 0 M; 15-185 min, 0 →0.6 M; 185-190 min, 0.6 →1.0 M) in 20 mM Tris-HCl (pH 8.0). The flow rate was 1 ml/min. Eluted oligo/ polySia-DMBs were monitored with fluorescent detector (Ex 373 nm, Em 448 nm). (A) Chromatograms of MH-FAEC. Colominic acid, PEB and PAB were analyzed. Three independent experiments were performed and typical chromatograms were shown. The average of Maximum DP (B) and the relative amounts of oligo/polySia ((Neu5Ac) n-DMB, n ≥ 6) obtained by three independent experiments. The sum of intensity of oligo/polySia (Neu5Ac) n-DMB, n ≥ 6) was set to 1.0.