HPLC Analysis of Horny Goat Weed Extract on Ascentis^® Express C18

sample preparation

to 100 mg of Horny Goat Weed herbal supplement add 10 mL ethanol, sonicate for 30 min, centrifuge 10,000 rpm for 1 min, filter supernatant using 0.45 μm filter, dilute to 30:70, water:ethanol

column

Ascentis Express C18, 15 cm x 4.6 mm I.D., 5 μm particles (50537-U)

mobile phase

(A) 0.1% acetic acid in water; (B) 0.1% acetic acid in acetonitrile;

gradient

5% B for 0.75 min; to 40% B in 3.75 min; to 100% B in 3.5 min; held at 100% B for 2 min

flow rate

1.5 mL/min

pressure

2120 psi (146 bar)

column temp.

35 °C

detector

UV, 250 nm

injection

5 μL

sample

each compound, 50 μg/mL in water:methanol (70:30)