Download PDF
Planta Med 2005; 71(9): 841-846
DOI: 10.1055/s-2005-871230
Original Paper
Biochemistry and Molecular Biology
© Georg Thieme Verlag KG Stuttgart · New York

Identification of Botanicals and Potential Contaminants through RFLP and Sequencing

Michelle R. Lum1 , 4 , Ellie Potter1 , 4 , Thanh Dang1 , David Heber2 , Mary Hardy2 , Ann M. Hirsch1 , 3
  • 1Department of Molecular, Cell and Developmental Biology, University of California, Los Angeles, CA, USA
  • 2Center for Human Nutrition, School of Medicine, University of California, Los Angeles, CA, USA
  • 3Molecular Biology Institute, University of California, Los Angeles, CA, USA
  • 4These authors should both be considered jointly as first author
Further Information

Publication History

Received: November 9, 2004

Accepted: March 30, 2005

Publication Date:
29 July 2005 (online)

    Permissions and Reprints

Abstract

Botanical supplements for health enhancement are being increasingly used in the United States, but no safeguards are formally in place to ensure that they are not contaminated with non-efficacious or potentially harmful plant material. A molecular approach, which allows the authentication of botanical ingredients and detection of contaminating plant material by analyzing the ITS-1 region by PCR-RFLP and subsequent sequencing, is described. When using starting material from which DNA can be obtained, this method has the potential for identifying both primary and contaminating plant material in botanical dietary supplements.

Key words

PCR amplification - alfalfa - red clover - repair reaction - RFLP - dietary supplements

References

  • 1 Slifman N R, Obermeyer W R, Aloi B K, Musser S M, Correll W A, Cichowicz S M. et al . Contamination of botanical dietary supplements by Digitalis lanata .  New Eng J Med. 1998;  339 806-11
    CrossrefPubMedGoogle Scholar
  • 2 Vanherweghem J -L, Depierreux M; Tielemans C, Abramowicz D, Dratwa M, Jadoul M. et al . Rapidly progressive interstitial fibrosis in young women: association with slimming regimen including Chinese herbs.  Lancet. 1993;  341 387-91
    CrossrefPubMedGoogle Scholar
  • 3 Vanherweghem J -L. Misuse of herbal remedies: the case of an outbreak of terminal renal failure in Belgium (Chinese herbs nephropathy.) .  J Altern Complement Med. 1998;  4 9-13
    PubMedGoogle Scholar
  • 4 Frasca T, Brett A S, Yoo S D. Mandrake toxicity. A case of mistaken identity.  Arch Intern Med. 1997;  157 2007-9
    CrossrefPubMedGoogle Scholar
  • 5 Quality control methods for medicinal plant materials. Geneva; World Health Organization 1998
    Google Scholar
  • 6 Brisken D P, Leroy A, Gawienowski M E. Influence of nitrogen on the production of hypericins by St. John’s wort.  Plant Physiol Biochem. 2000;  38 1-8
    CrossrefPubMedGoogle Scholar
  • 7 Pusch C M, Giddings I, Scholz M. Repair of degraded duplex DNA from prehistoric samples using Escherichia coli DNA polymerase I and T4 DNA ligase.  Nucleic Acids Res. 1998;  26 857-9
    CrossrefPubMedGoogle Scholar
  • 8 LeRoy A, Potter E, Woo H -Y, Heber D, Hirsch A M. Characterization and identification of alfalfa and red clover dietary supplements using a PCR-based method.  J Agric Food Chem. 2002;  50 5063-9
    CrossrefPubMedGoogle Scholar
  • 9 Baldwin B, Sanderson M, Porter J, Wojciechowski M, Campbell C, Donoghue M. The ITS region of nuclear ribosomal DNA: a valuable source of evidence on angiosperm phylogeny.  Ann Missouri Bot Garden. 1995;  82 247-277
    CrossrefPubMedGoogle Scholar
  • 10 Blattner F R. Direct amplification of the entire ITS region from poorly preserved plant material using recombinant PCR.  Biotechniques. 1999;  27 1180-6
    PubMedGoogle Scholar
  • 11 Altschul S F, Gish W, Miller W, Myers E W, Lipman D J. Basic local alignment search tool.  J Mol Biol. 1990;  215 403-10
    CrossrefPubMedGoogle Scholar
  • 12 Thompson J D, Higgins D G, Gibson T J. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position specific gap penalties and weight matrix choice.  Nucleic Acids Res. 1994;  22 4673-80
    PubMedGoogle Scholar
  • 13 Mihalov J J, Marderosian A D, Pierce J C. DNA identification of commercial ginseng samples.  J Agric Food Chem. 2000;  48 3744-52
    CrossrefPubMedGoogle Scholar
  • 14 Zhang K TB, Leung H W, Yeung H W, Wong R NS. Differentiation of Lycium barbarum from its related Lycium species using random amplified polymorphic DNA.  Planta Med. 2001;  67 379-81
    Article in Thieme ConnectPubMedGoogle Scholar
  • 15 Cheng K T, Tsay H S, Chen C F, Chou T W. Determination of the components in a Chinese prescription, Yu-Ping-Feng San, by RAPD analysis.  Planta Med. 1998;  64 563-5
    PubMedGoogle Scholar
  • 16 Ha W -Y, Yau C -F, Shaw P -C, Wang J. Differentiation of Panax ginseng from P. quinquefolius by Amplified Fragment Length Polymorphism. In: Shaw PC, Wang J, But PP-H, editors Authentication of Chinese Medicinal Materials by DNA Technology. Singapore; World Scientific 2002: pp 109-24
    Google Scholar
  • 17 Ngan F, Shaw P, But P, Wang J. Molecular authentication of Panax species.  Phytochemistry. 1999;  50 787-91
    CrossrefPubMedGoogle Scholar
  • 18 Fu R -Z, Wang J, Zhang Y -B, Wang Z -T, But P P-H, Li N. et al . Differentiation of medicinal Codonopsis species from adulterants by polymerase chain reaction-restriction fragment length polymorphism.  Planta Med. 1999;  65 648-50
    Article in Thieme ConnectPubMedGoogle Scholar
  • 19 Hillis D, Dixon M. Ribosomal DNA: Molecular evolution and phylogenetic inference.  Q Rev Biol. 1991;  66 411-53
    PubMedGoogle Scholar

Ann M. Hirsch

Department of Molecular, Cell and Developmental Biology

University of California

Los Angeles

CA 90095-1606

USA

Phone: +1-310-206-8673

Fax: +1-310-206-5413

Email: ahirsch@ucla.edu

      >