Objective: To establish a HPLC-DAD model for the simultaneous determination of two selaginellins (selaginellin and selaginellin B) and four biflavones (amentoflavone, sequoiaflavone, hinokiflavone and isocryptomerin) contained in 10 batches of Selaginella tamariscina and 12 batches of S. pulvinata produced in different areas.
Method: The analysis was performed on a Waters Cosmosil C18 column (4.6 mm x 250 mm, 5 microm) eluted with acetonitril-0.1% formic acid as mobile phase in a linear gradient mode. The detection wavelength was set at 280, 337 nm. The flow rate was kept at 1.0 mL x min(-1), and the column temperature was 30 degrees C.
Result: The six active constituents showed significant different in content. Amentoflavone in S. tamariscina contains (5. 628-9. 184 mg x g(-1)) is more than that contained in S. pulvinata (0.823-7.131 mg x g(-1)), while selaginellin in S. pulvinata (0.123-0.593 mg x g(-1)) is more than that contained in S. tamariscina (0.067-0.133 mg x g(-1)). All the calibration curves showed good linear correlation coefficients (r > 0.9997) over the wide test ranges.
Conclusion: The developed HPLC-DAD method is simple, sensitive and accurate and has the good repeatability in separation, which is available for the quality control of S. tamariscina and S. pulvinata.