Abstract

A reversed phase high performance liquid chromatography (RP-HPLC) method was described for simultaneous determination of seven effective constituents (orientin, isoorientin, vitexin, isovitexin, chlorogenic acid, caffeic acid and ferulic acid) in Bamboo leaf from nine cultivars of Phyllostachys pubescens. The seven compounds were simultaneously determined by HPLC with Sunfire C₁₈ ODS column (4.6 × 250 mm, 5 μm) by gradient elution using acetonitrile (A) and 0.8% acetic acid (B) as the mobile phase. The flow rate was 1.0 mL·min^-1; the detection wavelength was 335 nm with column temperature at 25°C. Finally, these effective components were separated clearly and respectively in 28 min, the linear ranges of orientin, isoorientin, vitexin, isovitexin,chlorogenic acid, caffeic acid and ferulic acid were 0.093 to 0.623 μg (r ＝ 0.9994), 0.058 to 0.385 μg (r ＝ 0.9994), 0.045 to 0.301 μg (r ＝ 0.9987), 0.041 to 0.272 μg (r ＝ 0.9992), 0.059 to 0.396 μg (r ＝ 0.9997), 0.046 to 0.312 μg (r ＝ 0.9992), 0.103 to 0.684 μg (r ＝ 0.9983), respectively; The average recoveries (n = 3) were 98.75, 98.34, 98.32, 99.70, 98.61, 98.20 and 99.31%, respectively. The validated method is simple, accurate and can be widely applied to quantification of the four flavonoids and three phenolic acid compounds in 9 different cultivars of P. pubescens.

Key words: Reversed phase high performance liquid chromatography (RP-HPLC), orientin, isoorientin, vitexin, isovitexin, chlorogenic acid, caffeic acid, ferulic acid.