Azide → Sinefungin

Participants

Reaction Steps

To a mixture consisting of Compound 8 (<671 μg, <0.0016 mmol) in water (100 μL) with sodium ascorbate (1 mg, 0.005 mmol) in water (200 μL) was added copper(II) sulfate (500 μg, 0.0031 mmol) in water (100 μL) and CAL Fluor® Red 610 azide (Biosearch Technologies, Inc. Lot #MVR1716, 1.4 mg, 0.0016 mmol) in DMF (400 μL). The reaction mixture was stirred at room temperature overnight. HPLC (Gemini C18, 110 {acute over (Å)}, 5 {acute over (Å)}, 250×4.6 mm, 1 mL/min, 5/95/0.05→90/10/0.05 ACN/H2O/TFA) showed mainly one product (13.1 min, 3%) and starting fluorophore azide (19.4 min, 75%). Prep HPLC (Gemini C18, 110 {acute over (Å)}, 10μ, 250×21.2 mm, 20 mL/min, 15/85/0.05→90/10/0.05 ACN/H2O/TFA) was done to purify the product which eluted from the column at around 8.6 min. The product solution was concentrated to remove the ACN and then lyophilized to afford the title detection analyte compound (12 μg as quantified by UV (MeOH, extinction coefficient 91,000), ˜0.6% yield) as a dark purple solid; LC/MS (Gemini C18, 3μ, 2.0×50 mm, 400 μL/min, A: 90/10/0.01 H2O/MeOH/HOAc, B 90/10/0.01 MeOH/H2O/HOAc 0-6 min 0-100% B, 6-9 hold at 100% B, 9.1-15 min re-equilibrate at 0% B) consistent with that expected for pure title detection analyte compound (5.1 min, ESI+ m/z=1162/2=581).

Provenance & Evidence