Pharmaceutical Medicine

, Volume 32, Issue 6, pp 429–444 | Cite as

Adulterants and Contaminants in Psychotropic Herbal Medicines Detected with Mass Spectrometry and Next-Generation DNA Sequencing

  • Claire L. HobanEmail author
  • Ian F. Musgrave
  • Megan L. Coghlan
  • Matthew W. P. Power
  • Roger W. Byard
  • Christine Nash
  • Rachael Farrington
  • Garth Maker
  • Elly Crighton
  • Robert Trengove
  • Michael Bunce
Original Research Article



The role of herbal medicine in the treatment of common psychiatric disorders such as anxiety, depression and insomnia has become more established over the past decade. Some herbal preparations such as St John’s wort (Hypericum perforatum) have demonstrated clinical evidence but have also been included in recent reports of widespread adulteration and contamination. Herbal medicines sold in Australia are required to be listed on the Therapeutic Goods Administration’s (TGA) Australian Register of Therapeutic Goods (ARTG) and must comply with strict ingredient and manufacturing guidelines to assure quality and safety.


The aim of this research was to assess whether pharmaceutical adulterants and contaminants were present in psychotropic herbal medicines available in Australia, as a measure of quality, and the effectiveness of regulation.


A two-pronged approach combining next-generation DNA sequencing and small-molecule analysis techniques was undertaken to audit a subset of herbal medicines for the presence of prescription medications, illicit drugs, pesticides, herbicides, heavy metals and contaminant DNA. Small-molecule analysis included liquid chromatography with quadrupole time-of-flight mass spectrometer (LC-QTOF-MS) detection, liquid chromatography with UV/vis diode array (LC-UV) detection, gas chromatography with nitrogen–phosphorus and mass spectrometer detection (GC-NPD/MS) and heavy metal analysis using inductively coupled plasma with mass spectrometer (ICP-MS) detection.


In total, 49% (29 of 59) of the investigated herbal medicines had one or more materials not listed on their labels or ARTG registration, including Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES)-listed material (one medicine), heavy metals (12%) or components that could trigger food sensitivity, such as wheat (12%). In contrast to previous studies, no prescription pharmaceutical adulterants were detected, although 10% had undeclared caffeine. Twenty-four percent of herbal medicines had DNA from animal species, including mice and bats, indicating poor quality control. The surveyed herbal medicines included both traditional Chinese medicines (TCM) and Western herbals. Ninety-four percent of TCMs were contaminated or adulterated, compared with 37% of the Western herbals. Only two of the 59 samples contained the listed active ingredient(s) without additional adulterants and contaminants, or missing ingredients.


The high levels of contamination found in this study suggests that closer surveillance of herbal medicines is needed in order to assure the required level of quality of herbal medicines available in Australia. The results suggest that the TGA’s low-/high-risk system for regulation coupled with post-market auditing is not keeping unapproved and/or unsafe herbal medicines from the market.


Compliance with Ethical Standards


This study was funded by the Australian National Health and Medical Research Council (NHMRC) (grant #1061329).

Conflict of interest

Claire L. Hoban, Ian F. Musgrave, Megan L. Coghlan, Matthew W.P. Power, Roger W. Byard, Christine Nash, Rachael Farrington, Garth Maker, Elly Crighton, Robert Trengove and Michael Bunce report no known conflicts of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

40290_2018_252_MOESM1_ESM.pdf (102 kb)
Supplementary material 1 (PDF 101 kb)


  1. 1.
    Sarris J. Herbal medicines in the treatment of psychiatric disorders: 10-year updated review. Phytother Res. 2018;32(7):1147–62.CrossRefGoogle Scholar
  2. 2.
    Coghlan ML, Maker G, Crighton E, Haile J, Murray DC, White NE, et al. Combined DNA, toxicological and heavy metal analyses provides an auditing toolkit to improve pharmacovigilance of traditional Chinese medicine (TCM). Sci Rep. 2015;10:17475.CrossRefGoogle Scholar
  3. 3.
    Newmaster SG, Grguric M, Shanmughanandhan D, Ramalingam S, Ragupathy S. DNA barcoding detects contamination and substitution in North American herbal products. BMC Med. 2013;11:222.CrossRefGoogle Scholar
  4. 4.
    Parveen I, Gafner S, Techen N, Murch SJ, Khan IA. DNA barcoding for the identification of botanicals in herbal medicine and dietary supplements: strengths and limitations. Planta Med. 2016;82(14):1225–35.CrossRefGoogle Scholar
  5. 5.
    Sarwat M, Yamdagni MM. DNA barcoding, microarrays and next generation sequencing: recent tools for genetic diversity estimation and authentication of medicinal plants. Crit Rev Biotechnol. 2016;36(2):191–203.CrossRefGoogle Scholar
  6. 6.
    Ivanova NV, Kuzmina ML, Braukmann TW, Borisenko AV, Zakharov EV. Authentication of herbal supplements using next-generation sequencing. PLoS One. 2016;11(5):e0156426.CrossRefGoogle Scholar
  7. 7.
    Mishra P, Kumar A, Nagireddy A, Mani DN, Shukla AK, et al. DNA barcoding: an efficient tool to overcome authentication challenges in the herbal market. Plant Biotechnol J. 2016;14(1):8–21.CrossRefGoogle Scholar
  8. 8.
    Blacksell L, Byard RW, Musgrave IF. Forensic problems with the composition and content of herbal medicines. J Forensic Leg Med. 2014;23:19–21.CrossRefGoogle Scholar
  9. 9.
    Hoban CL, Byard RW, Musgrave IF. A comparison of patterns of spontaneous adverse drug reaction reporting with St. John’s wort and fluoxetine during the period 2000–2013. Clin Exp Pharmacol Physiol. 2015;42:747–51.CrossRefGoogle Scholar
  10. 10.
    Gilbert JD, Musgrave IF, Hoban C, Byard RW. Lethal hepatocellular necrosis associated with herbal polypharmacy in a patient with chronic hepatitis B infection. Forensic Sci Int. 2014;24:138–40.CrossRefGoogle Scholar
  11. 11.
    Najafian J, Abdar-Esfahani M, Arab-Momeni M, Akhavan-Tabib A. Safety of herbal medicine in treatment of weight loss. ARYA Atheroscler. 2014;10:55–8.PubMedPubMedCentralGoogle Scholar
  12. 12.
    Kostakis C, Byard RW. Sudden death associated with intravenous injection of toad extract. Forensic Sci Int. 2009;188:e1–5.CrossRefGoogle Scholar
  13. 13.
    O’Callaghan C, Quine S. How older Vietnamese Australian women manage their medicines. J Cross Cult Gerontol. 2007;22:405–19.CrossRefGoogle Scholar
  14. 14.
    Sims DN, Felgate PD, Felgate HE, Lokan RJ. Application of a simple extraction procedure using aqueous ammonia to the analysis of basic drugs in blood by GC. Forensic Sci Int. 1991;49:33–42.CrossRefGoogle Scholar
  15. 15.
    Coghlan ML, Haile J, Houston J, Murray DC, White NE, et al. Deep sequencing of plant and animal DNA contained within traditional Chinese medicines reveals legality issues and health safety concerns. PLoS Genet. 2012;8:e1002657.CrossRefGoogle Scholar
  16. 16.
    Taberlet P, Coissac E, Pompanon F, Gielly L, Miquel C, et al. Power and limitations of the chloroplast trnL (UAA) intron for plant DNA barcoding. Nucl Acid Res. 2007;35:e14.CrossRefGoogle Scholar
  17. 17.
    Taylor PG. Reproducibility of ancient DNA sequences from extinct Pleistocene fauna. Mol Biol Evol. 1996;13(1):283–5.CrossRefGoogle Scholar
  18. 18.
    Murray DC, Coghlan ML, Bunce M. From benchtop to desktop: important considerations when designing amplicon sequencing workflows. PLoS ONE. 2015;10(4):e0124671.CrossRefGoogle Scholar
  19. 19.
    Poinar HN, Hofreiter M, Spaulding WG, Martin PS, Stankiewicz BA, et al. Molecular coproscopy: dung and diet of the extinct ground sloth Nothrotheriops shastensis. Science. 1998;281:402–6.CrossRefGoogle Scholar
  20. 20.
    Kearse M, Moir R, Wilson A, Stones-Havas S, Cheung M, et al. Geneious Basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinf. 2012;28(12):1647–9.CrossRefGoogle Scholar
  21. 21.
    Edgar RC. Search and clustering orders of magnitude faster than BLAST. Bioinf. 2010;26:2460–1.CrossRefGoogle Scholar
  22. 22.
    Edgar RC, Haas BJ, Clemente JC, Quince C, Knight R. UCHIME improves sensitivity and speed of chimera detection. Bioinf. 2011;27:2194–200.CrossRefGoogle Scholar
  23. 23.
    Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic local alignment search tool. J Mol Biol. 1990;215:403–10.CrossRefGoogle Scholar
  24. 24.
    Huson DH, Auch AF, Qi J, Schuster SC. MEGAN analysis of metagenomic data. Genome Res. 2007;17(3):377–86.CrossRefGoogle Scholar
  25. 25.
    Therapeutic Goods Administration, Australian Regulatory Guidelines, Good Manufacturing Practice (GMP) clearance for overseas manufacturers. 17th ed. Version 1.0 May 2011. Accessed 22 Jun 2017.
  26. 26.
    Grantley J, McPherson F, Petroeschevsky A. Recognising water weeds: plant identification guide. The State of New South Wales Industry & Investment; 2009. Accessed 22 Jun 2017.
  27. 27.
    Real A, Comino I, de Lorenzo L, Merchán F, Gil-Humanes J, et al. Molecular and immunological characterization of gluten proteins isolated from oat cultivars that differ in toxicity for celiac disease. PLoS ONE. 2012;7:e48365.CrossRefGoogle Scholar
  28. 28.
    Convention on International Trade in Endangered Species of Flora and Fauna, The CITES Appendices. Accessed Jul 2016.
  29. 29.
    Australian Government. Therapeutic Goods Order No. 69 general requirements for labels for medicines (27/08/2001)Google Scholar
  30. 30.
    Johnston A, Holt DW. Substandard drugs: a potential crisis for public health. Br J Clin Pharmacol. 2014;78(2):218–43.CrossRefGoogle Scholar
  31. 31.
    Calisher CH, Childs JE, Field HE, Holmes KV, Schountz T. Bats: important reservoir hosts of emerging viruses. Clin Microbiol Rev. 2006;19:531–45.CrossRefGoogle Scholar
  32. 32.
    Churchill S. Australian bats. 2nd ed. Crows Nest: Allen & Unwin; 2008.Google Scholar
  33. 33.
    Zhou J, Xie G, Yan X. Encyclopedia of traditional Chinese medicines. Molecular structures, pharmacological activities, natural sources and applications. Berlin: Springer; 2011.Google Scholar
  34. 34.
    Byard RW. Traditional medicines and species extinction: another side to forensic wildlife investigation. Forensic Sci Med Pathol. 2016;12:125–7.CrossRefGoogle Scholar
  35. 35.
    Allison DB, Cutter G, Poehlman ET, Moore DR, Barnes S. Exactly which synephrine alkaloids does Citrus aurantium (bitter orange) contain? Int J Obes (Lond). 2005;29:443–6.CrossRefGoogle Scholar
  36. 36.
    Arbo MD, Larentis ER, Linck VM, Aboy AL, Pimentel AL, Henriques AT, et al. Concentrations of p-synephrine in fruits and leaves of Citrus species (Rutaceae) and the acute toxicity testing of Citrus aurantium extract and p-synephrine. Food Chem Toxicol. 2008;46:2770–5.CrossRefGoogle Scholar
  37. 37.
    Musgrave IF, Farrington RL, Hoban CL, Byard RW. Caffeine toxicity in forensic practice: possible effects and under-appreciated sources. Forensic Sci Med Pathol. 2016;12(3):299–303.CrossRefGoogle Scholar
  38. 38.
    Pendleton M, Brown S, Thomas CM, Odle B. Potential toxicity of caffeine when used as a dietary supplement for weight loss. J Diet Suppl. 2013;10(1):1–5.CrossRefGoogle Scholar
  39. 39.
    Ching CK, Chen SPL, Lee HHC, Lam YH, Ng SW, et al. Adulteration of proprietary Chinese medicines and health products with undeclared drugs: experience of a tertiary toxicology laboratory in Hong Kong. Br J Clin Pharmacol. 2018;84(1):172–8.CrossRefGoogle Scholar
  40. 40.
    Morales ME, Derbes RS, Ade CM, Ortego JC, Stark J, et al. Heavy metal exposure influences double strand break DNA repair outcomes. PLoS One. 2016;11(3):e0151367.CrossRefGoogle Scholar
  41. 41.
    Vickers KA, Jolly KB, Greenfield SM. Herbal medicine: women’s views, knowledge and interaction with doctors: a qualitative study. BMC Compl Altern Med. 2006;6:40.CrossRefGoogle Scholar
  42. 42.
    Blödt S, Mittring N, Schützler L, Fischer F, Holmberg C, et al. A consultation training program for physicians for communication about complementary medicine with breast cancer patients: a prospective, multi-center, cluster-randomized, mixed-method pilot study. BMC Cancer. 2016;16:843.CrossRefGoogle Scholar
  43. 43.
    MacLennan AH, Myers SP, Taylor AW. The continuing use of complementary and alternative medicine in South Australia: costs and beliefs in 2004. Med J Aust. 2006;184:27–31.PubMedGoogle Scholar
  44. 44.
    Brazier NC, Levine MAH. Drug-herb interaction among commonly used conventional medicines: a compendium for health care professionals. Am J Therap. 2003;10:163–9.CrossRefGoogle Scholar
  45. 45.
    Byard RW. The potential forensic significance of traditional herbal medicines. J Forensic Sci. 2010;55:89–92.CrossRefGoogle Scholar
  46. 46.
    de Boer HJ, Ichim MC, Newmaster SG. DNA barcoding and pharmacovigilance of herbal medicines. Drug Saf. 2015;38:611–20.CrossRefGoogle Scholar
  47. 47.
    McIntyre E, Saliba AJ, Wiener KK, Sarris J. Herbal medicine use behaviour in Australian adults who experience anxiety: a descriptive study. BMC Complement Altern Med. 2016;16:60–72.CrossRefGoogle Scholar
  48. 48.
    McIntyre E, Adams J, Foley H, Harnett J, Leach MJ. Consultations with naturopaths and Western herbalists prevalence of use and characteristics of users in Australia. J Altern Complement Med. 2018. Scholar

Copyright information

© Springer Nature Switzerland AG 2018

Authors and Affiliations

  • Claire L. Hoban
    • 1
    Email author
  • Ian F. Musgrave
    • 1
  • Megan L. Coghlan
    • 3
  • Matthew W. P. Power
    • 3
  • Roger W. Byard
    • 1
    • 2
  • Christine Nash
    • 2
  • Rachael Farrington
    • 1
  • Garth Maker
    • 4
    • 5
  • Elly Crighton
    • 4
    • 5
  • Robert Trengove
    • 4
  • Michael Bunce
    • 3
  1. 1.Adelaide Medical SchoolThe University of AdelaideAdelaideAustralia
  2. 2.Forensic Science SAAdelaideAustralia
  3. 3.Trace and Environmental DNA Laboratory, Department of Environment and AgricultureCurtin UniversityBentleyAustralia
  4. 4.Separation Science and Metabolomics Laboratory and the Advanced Mass Spectrometry FacilityMurdoch UniversityMurdochAustralia
  5. 5.School of Veterinary and Life SciencesMurdoch UniversityMurdochAustralia

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