Simultaneous determination of icariin,tetrahydropalmatine and osthole in Zhuanggu Zhitong granules by RP-HPLC

Objective: To simultaneously determine the contents of icariin,tetrahydropalmatine and osthole in Zhuanggu Zhitong granules. Methods: RP-HPLC method was used. The determination was performed on a Symmetry C18column(150 mm × 3. 9 mm,5 μm) with mobile phase consisting of acetonitrile(A)-0. 1% phosphoric acid(B; adjusted pH to 6. 5 with triethylamine) in gradient elution(0-24 min,24. 5% A; 24-70 min,24. 5% A→57. 5% A). The flow rate was 0. 8 mL·min-1. The column temperature was 35 ℃. The detection wavelength was set at 270 nm in 0-25 min,282 nm in 25-55 min,and 322 nm in 55-70 min. Injection volume was 20 μL. Results: The linear ranges of icariin,tetrahydropalmatine and osthole were 0. 255-0. 561 mg · mL-1(r = 0. 9996),6. 40-14. 08 μg·mL-1(r = 0. 9998) and 0. 170-0. 510 mg·mL-1(r = 0. 9999),respectively. The average recoveries(n = 6) were 99. 19%(RSD = 1. 2%),99. 97%(RSD = 0. 89%) and 99. 71%(RSD = 1. 2%),respectively. Conclusion: The method can be used for the quality control of Zhuanggu Zhitong granules by methodology validation.