Table 6. LC separation gradient for the purification of compound 7 and 8 (method A and B) and methods C and D used for quantification.

Instrument Parameter	Method A preparative	Method B preparative	Method C Analytical	Method D Analytical
Column	Sunfire C18 10 μm 10 x 250 mm D	YMC-Pack pro C4, 3 μm 12 nm, 150 x 4.6 mm D	Agilent XBD-Eclipse C18, 4.6 x 150 mm D Poresize 5 μm	YMC-Pack Pro C4 150 x4.6 mmD, S-3 μm
Flowrate	1.5 mL/min	1.0 mL/min	0.9 mL/min	0.9 mL/min
Injection Volume	200 μL	100 μL	20 μL	20 μL
Column temperature	30 °C	30 °C	35 °C	35 °C
Solvent A	milliQ water	milliQ water	0.1 % (v/v) Formic acid in water	0.1 % (v/v) Formic acid in water
Solvent B	Acetonitrile	Acetonitrile	0.1 % (v/v) Formic acid in Methanol	0.1 % (v/v) Formic acid in Methanol
Gradient	0- 20 min: 0 – 100 % B 20 – 32 min 100 % B	0- 25 min: 0 – 100 % B 20 – 32 min 100 % B	0 -25 min: 0 – 100 % B 25 – 35 min 100 % B	0 – 20 min: 10 – 100 % B 20 – 25 min 100 % B