Simultaneous Determination of Seven Active Components in Mian’anning Mixture by HPLC Under Multiple UV Wavelengths

OBJECTIVE To develop an HPLC method with multiple wavelength ultraviolet for the simultaneous determination the content of 5-hydroxymethyl-2-furaldehyde, polygalaxanthone Ⅲ, atractylenolide Ⅰ, atractylenolide Ⅲ, salvianolic acid B, tanshinone ⅡA and catalpol in Mian’anning mixture. METHODS The sample was separated on a Hypersil C₁₈ column(4.6 mm×250 mm, 5 μm) with a mobile phase consisting of methanol-acetonitrile(1∶3) and 0.1% phosphoric acid solution. The flow rate was 0.7 mL·min^-1. The detection wavelength was set at 284 nm for 5-hydroxymethyl-2-furaldehyde, 320 nm for polygalaxanthone Ⅲ, 220 nm for atractylenolide Ⅰ and atractylenolide Ⅲ, 270 nm for salvianolic acid B and tanshinone ⅡA, 210 nm for catalpol. The column temperature was 25 ℃. RESULTS The calibration curves of 5-hydroxymethyl-2-furaldehyde, polygalaxanthone Ⅲ, atractylenolide Ⅰ, atractylenolide Ⅲ, salvianolic acid B, tanshinone ⅡA and catalpol were linear in the range of 10.08-100.80 μg·mL^-1(r=0.999 8), 3.55-35.50 μg·mL^-1(r=0.999 3), 5.80-58.00 μg·mL^-1(r=0.999 1), 4.20-42.00 μg·mL^-1(r=0.999 9), 40.50-405.00 μg·mL^-1(r=1.000), 12.28-122.80 μg·mL^-1(r=0.999 5), 7.90-79.00 μg·mL^-1(r=0.999 9), and the average recoveries(n=6) were 98.56%(RSD=0.97%), 96.94%(RSD=1.73%), 99.61% (RSD=1.03%), 97.54%(RSD=1.35%), 100.04%(RSD=0.86%), 98.59%(RSD=1.70%), 99.81%(RSD=1.43%), respectively. CONCLUSION The method is feasible, accurate and stable, which can control the quatity of Mian’anning Mixture.