Elsevier

Analytica Chimica Acta

Volume 555, Issue 2, 12 January 2006, Pages 217-224
Analytica Chimica Acta

Multiple chromatographic fingerprinting and its application to the quality control of herbal medicines

https://doi.org/10.1016/j.aca.2005.09.037Get rights and content

Abstract

Recently, chromatographic fingerprinting has become one of the most powerful approaches to quality control of herbal medicines. However, the performance of reported chromatographic fingerprinting constructed by single chromatogram sometimes turns out to be inadequate for complex herbal medicines, such as multi-herb botanical drug products. In this study, multiple chromatographic fingerprinting, which consists of more than one chromatographic fingerprint and represents the whole characteristics of chemical constitutions of the complex medicine, is proposed as a potential strategy in this complicated case. As a typical example, a binary chromatographic fingerprinting of “Danshen Dropping Pill” (DSDP), the best-sold traditional Chinese medicine in China, was developed. First, two HPLC fingerprints that, respectively, represent chemical characteristics of depsides and saponins of DSDP were developed, which were used to construct binary chromatographic fingerprints of DSDP. Moreover, the authentication and validation of the binary fingerprints were performed. Then, a data-level information fusion method was employed to capture the chemical information encoded in two chromatographic fingerprints. Based on the fusion results, the lot-to-lot consistency and frauds can be determined either using similarity measure or by chemometrics approach. The application of binary chromatographic fingerprinting to consistency assessment and frauds detection of DSDP clearly demonstrated that the proposed method was a powerful approach to quality control of complex herbal medicines.

Introduction

With tremendous expansion in the use of herbal medicines worldwide, their quality control has been an important concern for both health authorities and the public [1], [2]. Among a variety of quality control methods, chromatographic fingerprinting has gained more and more attention recently. It has been widely introduced and accepted by WHO [3], FDA [4], EMEA [5], German Commission E [6], British Herbal Medicine Association [7], Indian Drug Manufacturers’ Association [8], and some other official or nonofficial organizations as a strategy for the assessment of herbal medicines. Lately, Chinese manufacturers are also required by Chinese State Food and Drug Administration (SFDA) to standardize their botanical injections using chromatographic fingerprinting [9].

Chromatographic fingerprint is a chromatogram that represents the chemical characteristics of herbal medicine [10]. Generally, samples with similar chromatographic fingerprint have similar properties. As a result, chromatographic fingerprinting has potential to determine the identity, authenticity, and lot-to-lot consistency of herbal medicines. So far, there are lots of chromatographic fingerprints reports of herbal medicines such as Ginkgo biloba[11], [12], [13], [14], Rhizoma chuanxiong[13], Salvia miltiorrhiza Bunge [15], [16], Angelica sinensis (Oliv.) diels[17], Forsythia suspensa (Thunb.) Vahl[18], Flos Carthami[19], Shenmai injection[10], [20], Tianjihuang[21], Cassia bark[22], etc. Meanwhile, high performance thin layer chromatography (HPTLC) [14], gas chromatography (GC) [18], high performance liquid chromatography (HPLC) [10], [11], [12], [17], [21], [22], capillary electrophoresis (CE) [19], high-speed counter-current chromatography (HSCCC) [15], [16], and some hyphenated chromatographic approaches are already employed to develop fingerprints. In all previous reports, chromatographic fingerprints for testing lot-to-lot consistency of herbal medicines were constructed by a single chromatogram. It was inadequate to represent all chemical patterns or characteristics when the compositions of the herbal medicine are too complex, e.g. multi-herb botanical drug product. It is almost impossible to develop appropriate analytical method (including samples preparation and chromatographic procedure) to represent all chemical characteristics of constituents in a chromatogram. In this complicated case, a combination of analytical methods with different separation principles and test conditions is recommended by FDA [4]. SFDA also indicates that combined chromatographic fingerprints might be useful [9]. Consequently, it is necessary to develop “multiple chromatographic fingerprints” for complex herbal medicine, which consist of more than one fingerprint and represent the whole characteristics of chemical constitutions of the complex medicine, to evaluate their quality, though obtaining reliable multiple chromatographic fingerprints and assessing the quality of herbal medicines through their multiple fingerprints are not trivial work.

In this study, “Danshen Dropping Pill” (DSDP), which is composed of S. miltiorrhiza Bunge (Chinese Danshen) and Panax notoginseng (Chinese Sanqi), was investigated as a typical example, to develop multiple chromatographic fingerprinting for quality control. DSDP is the most popular traditional Chinese medicine for the prevention and treatment of coronary arteriosclerosis, angina pectoris, and hyperlipaemia [23], [24], [25], [26], and has also been well sold as a diet supplement or a drug in a number of countries such as the USA, Russia, Singapore, South Korea, and UAE. In general, depsides and saponins, extracted from S. miltiorrhiza Bunge and P. notoginseng, respectively, are considered as the major active constituents of DSDP. Nevertheless, danshensu, a kind of depside, is the unique marking compound for the evaluation of the quality of this formula currently, which is set to at least 0.13 mg of danshensu per pill [23]. Apparently, this standard is not sufficient for quality control. To improve the standard of quality control of DSDP, chromatographic fingerprinting is necessary. Although a number of analytical methods including HPLC [27], [28], [29], direct MS [30], HPLC/MS [31], [32] for S. miltiorrhiza Bunge or P. notoginseng have been reported, it is still difficult to represent the whole chemical characteristics of depsides and saponins of DSDP in a single chromatogram because of their significant difference in chemical properties. In the pilot study, we found that the major depsides can be clearly represented in a single chromatogram, but some of the major saponins, such as notoginsenoside R1, cannot be well represented in the chromatogram due to their low concentration and weak UV absorption. In this work, two HPLC fingerprints that, respectively, represent chemical characteristics of depsides and saponins in DSDP were developed, which constructed binary chromatographic fingerprints of DSDP. Further, the binary chromatographic fingerprints were applied to testing lot-to-lot consistency and detecting frauds. The proposed methods are equally applicable to other complex herbal medicines for quality control.

Section snippets

Methodology of multiple chromatographic fingerprinting

In general, multiple chromatographic fingerprinting is composed of the multiple chromatographic fingerprints acquirement procedure (including analytical methods, authentication, and analytical methods validation) and fingerprints comparison procedure.

In chromatographic fingerprints acquirement procedure, we are able to obtain multiple chromatograms, which demonstrate that they could chemically represent characteristics of the analyte, and validate the analytical methods. Moreover, chemical

Authentication of binary chromatographic fingerprints

The resulting chromatograms are shown in Fig. 1. The main constituents of DSDP samples were identified by UV spectra and LC/MS as described in Section 2.4.2. On the basis of these UV spectra, MS spectra of [M  H] ions and previous results of our lab [34] and others [27], [32], the main peaks of two chromatograms were identified in Table 1, Table 2. Some of the results were confirmed by comparison with the standards.

Obviously, the main constituents of DSDP in chromatogram I are depsides,

Conclusion

In this study, multiple chromatographic fingerprinting, which consists of more than one chromatographic fingerprint and represent the whole chemical characteristics of the analyte, is proposed as a strategy for quality control of complex herbal medicines instead of reported single chromatographic fingerprinting. As a typical example, binary chromatographic fingerprinting of DSDP was developed for consistency assessment and frauds detection. The results indicate that multiple chromatographic

Acknowledgements

This study was financially supported by the Chinese Key Technologies R&D Program (Grant No. 2001BA701A01) and a key grant from the National Natural Science Foundation of China (Grant No. 90209005). The authors would like to thank Dr. Leming Shi of the National Center for Toxicological Research of the US Food and Drug Administration for constructive suggestions and helpful discussions.

References (35)

  • P. Pietta et al.

    J. Chromatogr.

    (1991)
  • A. Hasler et al.

    J. Chromatogr.

    (1992)
  • F. Gong et al.

    J. Chromatogr. A

    (2003)
  • M. Gu et al.

    J. Chromatogr. A

    (2004)
  • M. Gu et al.

    J. Chromatogr. A

    (2004)
  • Y. Sun et al.

    J. Chromatogr. B

    (2003)
  • L.W. Yang et al.

    J. Chromatogr. A

    (2005)
  • X.Y. Ji et al.

    Life Sci.

    (2003)
  • F. Liao et al.

    Biorheology

    (1995)
  • A.E. Lau et al.

    J. Chromatogr. A

    (2003)
  • J.L. Zhang et al.

    J. Pharm. Biomed. Anal.

    (2005)
  • D. Normile

    Science

    (2003)
  • D.M. Eisenberg et al.

    J. Am. Med. Assoc.

    (1998)
  • General Guidelines for Methodologies on Research and Evaluation of Traditional Medicine, World Health Organization,...
  • FDA Guidance for Industry—Botanical Drug Products (Draft Guidance), US Food and Drug Administration,...
  • Final Proposals for Revision of the Note for Guidance on Quality of Herbal Remedies, EMEA,...
  • J.B. Calixto

    Braz. J. Med. Biol. Res.

    (2000)
  • Cited by (0)

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