[Optimization of the separation conditions of fingerprint for Rhubarb by high performance liquid chromatography].

Objective: To optimize the separation conditions of fingerprint for Rhubarb by high performance liquid chromatography and lay the foundation of studies on its fingerprint by high performance liquid chromatography.
Methods: The reversed-phase high performance liquid chromatographic method was adopted. The samples were extracted by ultrasound and separated by a linear gradient elution on Shim-pack CLC-ODS column (6.0 mm i.d. x 150 mm, 5 microm), and then, with the methanol-1 ml/L phosphoric acid solution as mobile phase, were detected at UV 254 nm and column temperature 40 degrees C. During the linear gradient elution program, the volume fraction of methanol in mobile phase changed as follows: 0 min-5 min, 20%; 5 min-25 min, from 20% to 50%; 25 min-35 min, from 50% to 80%; 35 min-60 min, from 80% to 90%; 60 min-70 min, 90%. The flow rate was 0.8 ml/min from 0 min to 35 min, 0.6 ml/min from 35 min to 50 min and 1.0 ml/min after 50 min.
Results: The separation conditions described above achieved good result in the aspects of separable effect, stability, sensitivity, and reproducibility.
Conclusion: The optimum separation conditions of fingerprint would be useful for establishing the method of fingerprint for Rhubarb by high performance liquid chromatography.