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- The reaction may be monitored by HPLC: column: YMC ODS S5 4.6 × 150 mm; solvent A: 10% MeOH/H2O/0.2% H3PO4; solvent B: 90% MeOH/H2O/0.2% H3PO4; start with 0% B, linear gradient to 100% B over 20 min; injection volume: 5μL; flow rate: 1 mL/min; detection: 220nm