Download PDF
Planta Med 2009; 75(8): 860-863
DOI: 10.1055/s-0029-1185447
Analytical Studies
Letter
© Georg Thieme Verlag KG Stuttgart · New York

Composition of Indigo naturalis

Inken Plitzko1 , Tobias Mohn1 , Natalie Sedlacek1 , Matthias Hamburger1
  • 1Institute of Pharmaceutical Biology, University of Basel, Basel, Switzerland
Further Information

Publication History

received September 19, 2008 revised January 19, 2009

accepted January 22, 2009

Publication Date:
18 March 2009 (online)

Also available at
    Permissions and Reprints

Abstract

A proposal for a European Pharmacopoeia monograph concerning Indigo naturalis has recently been published, whereby the indigo (1) and indirubin (2) content should be determined by HPLC‐UV. This method was tested, but problems were seen with the dosage of indigo due to poor solubility. A quantitative assay for indigo based on 1H‐NMR was developed as an alternative. The HPLC and qNMR assays were compared with eight Indigo naturalis samples. The HPLC assay consistently gave much lower indigo concentrations because solubility was the limiting factor in sample preparation. In one sample, sucrose was identified by 1H‐NMR as an organic additive. Simple wet chemistry assays for undeclared additives such as sugars and starch were tested with artificially spiked Indigo naturalis samples to establish their limits of detection, and sulfate ash determinations were carried out in view of a better assessment of Indigo naturalis in a future European monograph.

Key words

Indigo naturalis - indigo - indirubin - qNMR - HPLC - Pharmacopoeia monograph

References

  • 1 Stöger E, Friedel F. Aktualisierung, Monographie „Indigo naturalis“. Arzneibuch der chinesischen Medizin. Stuttgart; Deutscher Apotheker Verlag 2006: 11
    Search in Google Scholar
  • 2 Chen D H, Xie J X. Chemical constituents of traditional Chinese medicine Qing Dai.  Chinese Tradit Herbal Drugs. 1984;  1 6-8
    PubMedSearch in Google Scholar
  • 3 Leclerc S, Garnier M, Hoessel R, Marko D, Bibb J A, Snyder G L, Greengard P, Biernat J, Wu Y Z, Mandelkow E M, Eisenbrand G, Meijer L. Indirubins inhibit glycogen synthase kinase-3β and CDK5/P25, two protein kinases involved in abnormal tau phosphorylation in Alzheimer's disease.  J Biol Chem. 2001;  276 251-260
    PubMedSearch in Google Scholar
  • 4 Hoessel R, Leclerc S, Endicott J A, Nobel M E, Lawrie A, Tunnah P, Leost M, Damiens E, Marie D, Marko D, Niederberger E, Tang W, Eisenbrand G, Meijer L. Indirubin, the active constituent of a Chinese antileukaemia medicine, inhibits cyclin-dependent kinases.  Nat Cell Biol. 1999;  1 60-67
    CrossrefPubMedSearch in Google Scholar
  • 5 [no authors] .Monograph of the Chinese Pharmacopoeia, Engl. edition. Beijing; The Pharmacopoeia Commission of PRC 2005
    Search in Google Scholar
  • 6 [no authors] . Monograph proposal „natural indigo“.  Pharmeuropa. 2008;  20 (1) 118
    PubMedSearch in Google Scholar
  • 7 Hamburger M. Isatis tinctoria – from the rediscovery of an ancient medicinal plant towards a novel anti-inflammatory phytopharmaceutical.  Phytochem Rev. 2002;  1 333-344
    CrossrefPubMedSearch in Google Scholar
  • 8 Oberthür C, Schneider B, Graf H, Hamburger M. The elusive indigo precursors in woad (Isatis tinctoria L.) – identification of the major indigo precursor, isatan A, and a structure revision of isatan B.  Chem Biodivers. 2004;  1 174-182
    PubMedSearch in Google Scholar
  • 9 Oberthür C, Graf H, Hamburger M. The content of indigo precursors in Isatis tinctoria leaves – a comparative study of selected accessions and post-harvest treatments.  Phytochemistry. 2004;  65 3261-3268
    CrossrefPubMedSearch in Google Scholar
  • 10 Mohn T, Hamburger M. Glucosinolate pattern in Isatis tinctoria and I. indigotica seeds.  Planta Med. 2008;  74 885-888
    Thieme ConnectPubMedSearch in Google Scholar
  • 11 Yee S K, Chu S S, Xu Y M, Choo P L. Regulatory control of Chinese proprietary medicines in Singapore.  Health Policy. 2005;  71 133-149
    CrossrefPubMedSearch in Google Scholar
  • 12 Benthin B, Danz H, Hamburger M. Pressurized liquid extraction of medicinal plants.  J Chromatogr A. 1999;  837 211-219
    CrossrefPubMedSearch in Google Scholar
  • 13 Basalo C, Mohn T, Hamburger M. Are extraction methods in quantitative assays of pharmacopoeia monographs exhaustive? A comparison with pressurized liquid extraction.  Planta Med. 2006;  72 1157-1162
    Thieme ConnectPubMedSearch in Google Scholar
  • 14 Pauli G F, Jaki B U, Lankin D C. A routine experimental protocol for qHNMR illustrated with taxol.  J Nat Prod. 2007;  70 589-595
    CrossrefPubMedSearch in Google Scholar
  • 15 Mohn T, Potterat O, Hamburger M. Quantification of active principles and pigments in leaf extracts of Isatis tinctoria by HPLC/UV/MS.  Planta Med. 2007;  73 151-156
    Thieme ConnectPubMedSearch in Google Scholar
  • 16 [no authors]. European Pharmacopeia, 6th edition. Strasbourg; Directorate for the quality of medicines of the Council of Europe 2008
    Search in Google Scholar
  • 17 Yang T, Wang Y, Xu F, Sun Q, Hu Y. Fermentation with microorganism for indigo.  Chin Tradit Patent Med. 2005;  27 914-915
    PubMedSearch in Google Scholar
  • 18 Wei Q, Yang M, Xu R, Sun Q, Hu Y. New conception about the processing of Indigo naturalis.  Chin Tradit Patent Med. 2004;  26 116-118
    PubMedSearch in Google Scholar
  • 19 Friedländer P, Roschdestwenski N. Über ein Oxidationsprodukt des Indigoblaus.  Ber Dtsch Chem Ges. 1915;  48 1841-1847
    CrossrefPubMedSearch in Google Scholar

Prof. Dr. Matthias Hamburger

Department of Pharmaceutical Sciences
Institute of Pharmaceutical Biology
University of Basel

Klingelbergstrasse 50

4056 Basel

Switzerland

Phone: + 41 6 12 67 14 25

Fax: + 41 6 12 67 14 74

Email: Matthias.Hamburger@unibas.ch

    www.thieme-connect.de/ejournals/toc/plantamedica