HPLC fingerprint analysis on Qibai Pingfei Granules and multi-components determination / 中草药

ABSTRACT

Objective: To establish an HPLC fingerprint of the compounds in Qibai Pingfei Granules (QPG), and to make a quantitative analysis. Methods: Sample was extracted by 50% methanol. Phenomenex Luna C18 column (250 mm × 4.6 mm, 5 μm) was used with a mobile phase of methanol-0.2% formic acid gradient elution. The flow rate was 1.0 mL/min, the detection wavelength was 250 nm, and the column temperature was 30℃. The chemical component fingerprint similarity of 10 batches of QPG was calculated with Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2012) published by National Pharmacopoeia Committee and the common peaks were identified by reference compounds. Results: Fingerprints of 10 batches of QPG were established and the similarities to the common mode were above 0.96. Totally 25 common peaks were found. Among them, peak 1 belonged to Pheretima, peaks 2 and 3 belonged to Pheretima and Ginseng Radix et Rhizoma (GRR), peak 4 belonged to GRR, Schisandrae Chinensis Fructus (SCF), and Chuanxiong Rhizoma (CR), peak 5 belonged to GRR, Allii Macrostemonis Bulbus (AMB), CR, and Pheretima, peaks 6, 8, 22, 23, 24, and 25 belonged to SCF, peak 7 belonged to Astragali Radix (AR) and SCF, peak 9 belonged to GRR and AMB, peak 10 belonged to Descurainiae Semen Lepidii Semen (DSLS) and AMB, peaks 11, 12, 13, 15, and 16 belonged to CR, peak 14 belonged to AR and DSLS, peaks 17, 18, 19, 20, and 21 belonged to AR. Based on the retention time, and UV absorption spectra of reference compounds, six constituents including caffeic acid (peak 12), ferulic acid (peak 13), schizandrol A (peak 22), schisantherin A (peak 23), deoxyschizandrin (peak 24), and schisandrin (peak 25) were identified. The linear ranges of caffeic acid, ferulic acid, schizandrol A, schisantherin A, deoxyschizandrin, and schisandrin were 3.38-108.02, 3.60-115.33, 2.99- 95.61, 2.81-89.77, 3.26-104.17, and 2.89-92.45 μg/mL, respectively. In 10 batches of QPG samples, the contents were as follows caffeic acid of 0.412-0.429 mg/g, ferulic acid of 0.302-0.317 mg/g, schizandrol of A 0.182-0.195 mg/g, schisantherin A of 0.179-0.195 mg/g, deoxyschizandrin of 0.203-0.215 mg/g, and schisandrin of 0.131-0.144 mg/g, the amount of each indicator composition among different batches changed a litte, and the sample quality is stable. Conclusion: The method has good precision, reproducibile, stability, separation, and can be used for the quality control of QPG.