Simultaneous determination of baicalin,chlorogenic acid and phillyrin in Shuang huanglian Oral Liquid by multiple wavelength RP-HPLC

AIM: To establish a method for simultaneously determining baicalin,chlorogenic acid and phillyrin in Shuanghuanglian Oral Liquid(Radix Scutellariae,Flos Lonicerae japonicac,Fructus Forsythiae). METHODS: A multiple wavelength HPLC method was devoloped.The analysis was performed on an Agilent Zorbax SB C_18 column(250 mm × 4.6 mm,5 μm) with linear gradient elution using 0.3% aqueous formic acid(A) and acetoniterile(B) in the order of 14%-20%/0-12 min,20%-30%/12-13 min,30%-65%/13-25 min and 65%/25-35 min.Baicalin,chlorogenic acid and phillyrin were detected at 280 nm,324 nm and 277 nm wavelength,respectively.The flow rate was 1.0 mL/min and the column temperature was 30 ℃. RESULTS: The calibration curves of baicalin,chlorogenic acid and phillyrin showed good linearity in the ranges of 0.225-3.60 μg,0.032 4-0.518 4 μg,0.135-2.16 μg,respectively.The average recoveries of them were 100.5 %,102.0 %,99.62% with RSD of 0.75%,3.9%,1.1%,respectively. CONCLUSION: The method is simple,accurate and reproducible for determining baicalin,chlorogenic acid and phillyrin in Shuanghuanglian Oral Liquid.