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85031229892
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For the stability measurements, a solution of 250 μg/mL of prodrug in 0.02 M phosphate buffer (pH 7.2) was incubated at 37°C. Aliquots (40 μL) were taken at various times and analyzed by HPLC after dilution with eluent (120 μL). HPLC conditions: analysis was carried out on a reverse-phase column (Spherisorb C18) using isocratic conditions (1 mL/min) of 60% phosphate buffer (0.02 M, pH 3) and 40% acetonitrile with UV detection at 265 nm.
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For the kinetic measurements, a solution of 300 μM of prodrug in 0.02 M phosphate buffer (pH 7.2) was incubated at 37°C in the presence of 10 units/mL of β-D-glucuronidase (E. coli). Aliquots (40 μL) were taken at various times and analyzed by HPLC after dilution with eluent (120 μL). HPLC conditions: analysis was carried out on a reverse-phase column (WATERS X-terra, 250/4.6 mm) using isocratic conditions (1 mL/min) of 70% triethylammonium acetate buffer (0.1%, pH 9) and 30% acetonitrile with UV detection at 265 nm (extracted from PDA 3-D spectra).
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