Chromatographic fingerprinting and quantitative analysis for the quality evaluation of Xinkeshu tablet

Abstract

A simple, sensitive and accurate method based on high performance liquid chromatography (HPLC) with diode array detector (DAD) was developed and validated for systematic quality evaluation of one type of traditional Chinese medicine preparations named Xinkeshu (XKS) tablet. In this study, the chromatographic fingerprints of XKS tablet were developed first, 23 peaks were selected as the common peaks to evaluate the similarities among different batches of XKS samples, which were manufactured in a long time span of three years. Additionally, simultaneous quantification of six markers in XKS tablet, including Danshensu, Protocatechuic aldehyde, Puerarin, Daidzin, Salvianolic acid B and Daidzein, was performed. The validation results showed that the developed method was specific, accurate, precise and robust. The preliminary explanation on why a close similarity between fingerprints did not exactly mean similar contents of chemical components in samples was given. The contribution of each chromatographic peak to similarity was also evaluated. The developed method offers an efficient, reliable and practical approach for systematic quality evaluation of XKS tablet.

Keywords: Fingerprint analysis; High-performance liquid chromatography; Multi-ingredient quantitative analysis; Quality evaluation; Xinkeshu tablet.

Figure 1

Molecular structures of the six compounds.

Figure 2

The HPLC–UV chromatogram of (A) 6 mixed standards and (B) the representative HPLC-fingerprint of extract of XKS. The numbers from 1 to 23 correspond to the common peaks.

Figure 3

HPLC fingerprints of 15 commercial samples of XKS from different batches and their standard fingerprint.

Figure 4

The contribution of each chromatographic peak to the similarity of the 15 batches of XKS. (A) and (B) refer to the difference of the $r^{\cos }$ or r before and after each peak was deleted in turn, and (C) refers to the average relative peak areas in the chromatographic fingerprints of the 15 batches of XKS (The peak of Puerarin was used as the reference peak).