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Determination of flavonoids in Hypericum perforatum by HPLC analysis
1Department of Pharmacognosy, China Pharmaceutical University, Nanjing 210038, China.
Aim:
To establish an HPLC method for simultaneous determination of four flavonoids in Hypericum perforatum L., ruitn, hyperin, avicularin and quercetin.
Methods:
C18 column was used, the chromatography was carried out with a linear gradient program. The mobile phase was A: H2O (pH 3.0-3.5 adjusted with phosphoric acid) and B: acetonitrile, at flow rate of 1.0 mL.min-1, peaks were detected at 254 nm.
Results:
The linear range of rutin was 1.376-8.256 micrograms.mL-1 (gamma = 0.9999), hyperin 3.160-18.960 micrograms.mL-1 (gamma = 0.9996), avicularin 0.968-5.808 micrograms.mL-1 (gamma = 0.9998) and quercetin 0.776-4.656 micrograms.mL-1 (gamma = 0.9993). The average recovery of rutin was 97.8%, RSD 4.8% (n = 3), hyperin 100.7%, RSD 3.7% (n = 3), avicularin 97.3%, RSD 0.8% (n = 3) and quercetin 100.5%, RSD 4.4% (n = 3). All of RSDs of precision were less than 2% (n = 5), reproducibilities less than 3%.
Conclusion:
The method is simple, effective and feasible. It can be used to determine flavonodis in Hypericum perforatum.