Simultaneous determination of nine kinds of active components in Hugan Agent by RP-HPLC

Hong-Sheng LIU

ABSTRACT

Objective:

To establish an RP-HPLC method for simultaneous determining the contents of nine kinds of components in Hugan Agent such as loganin, paeonflorin, scuteuarin, baicalin, baicalein, deoxyschizandrin, schisandrin, schizandrin C, and ursolic acid, and to provide the quality guarantee for it.

Methods:

Agilent Zorbax SB-C18 (150 mm×4.6 mm, 5 μm) column was used. Acetonitrile and 0.1% phosphoric acid water-solution were used as mobile phase of gradient elution, and the volume flow was at 1.0 mL/min; Ultraviolent determination wavelength was 236, 280, and 210 nm; Column temperature was at 30℃, and injection volume was 5 μL.

Results:

The lowest detection limit in loganin, paeonflorin, scuteuarin, baicalin, baicalein, deoxyschizandrin, schisandrin, schizandrin C, and ursolic acid respectively was 12.0, 2.60, 5.75, 9.75, 14.39, 19.06, 14.90, 15.63, and 16.08 ng; The linear range was 583.33-18.229, 916.67-28.65, 541.67-16.93, 416.67-13.02, 500.00-15.63, 458.33-14.32, 625.00-19.53, 458.33-14.32, and 1 000.00-31.25 mg/L, respectively; The average recovery in Hugan Agent was 103.51%, 104.19%, 95.16%, 96.71%, 105.61%, 96.12%, 97.09%, 96.87%, and 105.90%, respectively; The repeatability RSD was 1.39%, 1.41%, 1.87%, 1.91%, 1.79%, 1.45%, 1.32%, 1.71%, and 1.49%, respectively; The stability RSD was 1.36%, 1.22%, 1.87%, 1.91%, 1.93%, 1.56%, 1.39%, 1.78%, and 1.61%, respectively. 6 batche of Hugan Agent contained the average quality of loganin, ursolic acid, paeonflorin, scuteuarin, baicalin, baicalein deoxyschizandrin, schisandrin, and schizandrin respectively was 216.5-222.5, 40.8-42.8, 125.4-136.3, 144.0-147.3, 1640.8-1947.3, 497.5-515.0, 15.0-17.3, 33.6-36.0, and 3.0-3.9 mg/L.

Conclusion:

The method is believable for determining the concent of loganin, paeonflorin, scuteuarin, baicalin, baicalein, deoxyschizandrin, schisandrin, schizandrin C, and ursolic acid in Hugan Agent with its simplicity, sensibility, repeatability, and better recovery rate.