MULTIPLE COMPOUNDS DETERMINATION AND FINGERPRINT ANALYSIS OF PULSATILLA CHINENSIS (BUNGE) REGEL BY HPLC COUPLED WITH EVAPORATIVE LIGHT SCATTERING DETECTION FOR QUALITY CONTROL

Abstract

For comprehensive quality control of Pulsatilla chinensis (Bunge) Regel (Baitouweng in Chinese), the methods including multiple compounds determination (MCD) and chemical fingerprint analysis (CFA) were carried out. High-performance liquid chromatography (HPLC) coupled with evaporative light scattering detection (ELSD) was developed for analysis. In the process of MCD, methodology for analysis was evaluated and good results were obtained for simultaneous determination of the five marker compounds. In CFA, the similarities were calculated based on the relative peak areas (RPA) and relative retention times (RRT) of 18 characteristic peaks of the medicinal plants and two adulterants (Rhaponticum uniflorum (L.) DC and Potentilla discolor Bge) collected from different parts of China. Hierarchical cluster analysis (HCA) and principal component analysis (PCA) were effectively applied to distinguish the origins of different samples. The results indicate that the method by combination of MCD and CFA is suitable for quantitative analysis and quality control of P. chinensis (Bunge) Regel.

Keywords:

• chemical fingerprint analysis
• evaporative light scattering detection
• high-performance liquid chromatography
• Multiple compounds determination
• Pulsatilla Chinensis (Bunge) Regel
• quality control

ACKNOWLEDGMENTS

This work was supported financially by the Key Laboratory of University of Liaoning Provence (No. 2008S072) and the Science and Technology Foundation of Dalian (No. 2008E11SF 223), China. We express thanks to Dr. Yunpeng Diao (College of Pharmacy, Dalian Medical University, Dalian, China) for identification of the herbs collected from different locations.

Notes

^a y is the lg value of the peak area; x is the lg value of sample amount.

^b Recovery (%) = [(found amount-original amount)/spiked amount] × 100.

n.d.: not detected.

RRT: the ratio between peak retention time of target and internal reference substance.

n.d.: not detected.

RPA, the ratio between peak area of target and internal reference substance.

n.d.., not detected.

^a The reference fingerprint was developed with the median of all chromatograms.