HPLC Analysis of Withania Extract on Ascentis^® Express F5

column

Ascentis Express F5, 10 cm x 2.1 mm I.D., 2.7 μm particles (53569-U)

mobile phase

[A] water; [B] acetonitrile

gradient

20 to 100% B in 5 min; held at 100% B for 0.5 min

flow rate

0.3 mL/min

pressure

2750 psi (190 bar)

column temp.

35 °C

detector

UV, 227 nm

injection

5 μL

sample

20 mg/mL in water:acetonitrile (50:50); sonicate for 15 minutes; filter 0.45 μm; dilute to water:acetonitrile (80:20)