Study on simultaneous analysis of chlorogenic acid and flavonoid constituents in Flos Lonicerae by HPLC

OBJECTIVE To develop a RP-HPLC method with gradient elution for the separation and simultaneously determination of chlorogenic acid and four flavonoids, rutin, luteolin-7-O-β-D-galactoside, lonicerin and quercetin in Flos Lonicerae.METHODS The separation was performed on a Aglient Zorbax 80A column (4.6 mm×250 mm,5 μm). Acetonitril-0.5% H3PO4 was used as the mobile phase with gradient elution. The flow rate was 1.0 mL·min-1.The detection wavelength was at 355 nm. RESULTS Good linearities of chlorogenic acid, rutin, luteolin-7-O-β-D-galactoside, lonicerin and quercetin were obtained over the ranges of 0.275 - 22 μg ( r = 0.999 8 ) ,7.725 -772.5 ng(r = 0.9997),8.025 - 802.5 ng(r = 0.9999),7.725 - 772.5 ng (r = 0.999 9) and 3.338 - 333.8 ng( r = 0.999 9). The average recoveries(n = 9) were 99.42% ,99.11% , 101.46% ,98.61% and 100.65%, respectively. CONCLUSION This method is accurate , rapid and reproducible, will be used to determine chlorogenic acid and flavonoids in the raw material of Flos Lonicerae.