Download PDF
Planta Med 2022; 88(07): 538-547
DOI: 10.1055/a-1534-6928
Natural Product Chemistry and Analytical Studies
Original Papers

Quantitative Removal of Pyrrolizidine Alkaloids from Essential Oils by the Hydrodistillation Step in Their Manufacturing Process

David S. Giera
1   G. Pohl-Boskamp GmbH & Co. KG, Hohenlockstedt, Germany
,
Michael Preisitsch
2   PhytoLab GmbH & Co. KG, Vestenbergsgreuth, Germany
,
Hugues Brevard
3   Robertet SA, Grasse, France
,
Jörn Nemetz
1   G. Pohl-Boskamp GmbH & Co. KG, Hohenlockstedt, Germany
› Author Affiliations
› Further Information
    Permissions and Reprints

Abstract

Pyrrolizidine alkaloids are naturally occurring toxins produced by certain weeds that can, if accidentally co-harvested, contaminate plant-based food, feed, and herbal medicinal products. Focusing on herbal medicinal products, the presence of pyrrolizidine alkaloids is restricted by regulatory prescribed thresholds to assure patient safety. Among the multitude of different herbal active substances utilized in herbal medicinal products, the class of pharmaceutically effective essential oils is considered to exhibit a negligible contribution to pyrrolizidine alkaloid contamination. Within the present investigation, this hypothesis should be scientifically scrutinized. For this purpose, an experimental set-up was chosen that reproduces the typical manufacturing step of hydrodistillation. Essential oils of eucalyptus and lemon were selected exemplarily and spiked with 3 representative pyrrolizidine alkaloids (retrorsine, retrorsine-N-oxide, and lycopsamine), whereupon hydrodistillation was performed. Analysis of the resulting distillates by LC-MS/MS proved that artificially added pyrrolizidine alkaloids were removed completely. Moreover, quantitative pyrrolizidine alkaloid recovery in the aqueous phases was observed. Hence, it was experimentally confirmed that herbal medicinal products employing hydrodistilled essential oils of pharmaceutical quality are intrinsically free of pyrrolizidine alkaloids due to the particularities of their manufacturing process. Furthermore, it can be concluded from theoretical considerations that essential oils produced by cold pressing have a negligible risk of carrying pyrrolizidine alkaloid contamination. Our findings provide a strong indication that the requirement for analytical pyrrolizidine alkaloid testing of essential oils for pharmaceutical use should be fundamentally reconsidered.

Key words

pyrrolizidine alkaloids - herbal medicinal products - essential oils - hydrodistillation

Supporting Information

    A sample overview, as well as details on the preparation of spiked essential oil samples, analytics, and calculations, are available as Supporting Information.

  • Supporting Information


Publication History

Received: 16 December 2020

Accepted after revision: 22 June 2021

Article published online:
22 July 2021

© 2021. Thieme. All rights reserved.

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

 

  • References

  • 1 Brown ME. Survivors over six millennia: essential oils. Pharm Hist (Lond) 2014; 44: 13-18
    PubMedGoogle Scholar
  • 2 Feyaerts AF, Luyten W, Van Dijck P. Striking essential oil: tapping into a largely unexplored source for drug discovery. Sci Rep 2020; 10: 2867
    CrossrefPubMedGoogle Scholar
  • 3 Edris AE. Pharmaceutical and therapeutic potentials of essential oils and their individual volatile constituents: a review. Phytother Res 2007; 21: 308-323
    CrossrefPubMedGoogle Scholar
  • 4 Bakkali F, Averbeck S, Verbeck D, Idaomar M. Biological effects of essential oils – A review. Food Chem Toxicol 2008; 46: 446-475
    CrossrefPubMedGoogle Scholar
  • 5 Reichling J, Schnitzler P, Suschke U, Saller R. Essential oils of aromatic plants with antibacterial, antifungal, antiviral, and cytotoxic properties–an overview. Forsch Komplementmed 2009; 16: 79-90
    PubMedGoogle Scholar
  • 6 Angelini P, Tirillini B, Akhtar MS, Dimitriu L, Bricchi E, Bertuzzi G, Venanzoni R. Essential Oil with anticancer Activity: An Overview. In: Akhtar M, Swamy M. eds. Anticancer Plants: Natural Products and biotechnological Implements. Singapore: Springer; 2018: 207-231
    Google Scholar
  • 7 Fitsiou E, Pappa A. Anticancer activity of essential oils and other extracts from aromatic plants grown in Greece. Antioxidants 2019; 8: 290
    CrossrefPubMedGoogle Scholar
  • 8 Bayala B, Bassole IHN, Scifo R, Gnoula C, Morel L, Lobaccaro JMA, Simpore J. Anticancer activity of essential oils and their chemical components–a review. Am J Cancer Res 2014; 4: 591-607
    PubMedGoogle Scholar
  • 9 Lesgards JF, Baldovini N, Vidal N, Pietri S. Anticancer activities of essential oils constituents and synergy with conventional therapies: a review. Phytother Res 2014; 28: 1423-1446
    CrossrefPubMedGoogle Scholar
  • 10 Sobral MV, Xavier AL, Lima TC, de Sousa DP. Antitumor activity of monoterpenes found in essential oils. Sci World J 2014; 2014: 953451
    CrossrefPubMedGoogle Scholar
  • 11 Privitera G, Luca T, Castorina S, Passanisi R, Ruberto G, Napoli E. Anticancer activity of Salvia officinalis essential oil and its principal constituents against hormone-dependent tumour cells. Asian Pac J Trop Biomed 2019; 9: 24-28
    CrossrefPubMedGoogle Scholar
  • 12 Dahham SS, Hassan LEA, Ahamed MBK, Majid ASA, Majid AMSA, Zulkepli NN. In vivo toxicity and antitumor activity of essential oils extract from agarwood (Aquilaria crassna). BMC Complement. Altern Med 2016; 16: 326
    PubMedGoogle Scholar
  • 13 Nikolić M, Glamočlija J, Ferreira ICFR, Calhelha RC, Fernandes Â, Marković T, Marković D, Giweli A, Soković M. Chemical composition, antimicrobial, antioxidant and antitumor activity of Thymus serpyllum L., Thymus algeriensis Boiss. and Reut and Thymus vulgaris L. essential oils. Ind Crops Prod 2014; 52: 183-190
    CrossrefPubMedGoogle Scholar
  • 14 Hensel A, Bauer R, Heinrich M, Spiegler V, Kayser O, Hempel G, Kraft K. Challenges at the time of COVID-19: opportunities and innovations in antivirals from nature. Planta Med 2020; 86: 659-664
    Article in Thieme ConnectPubMedGoogle Scholar
  • 15 Thomsen J, Röschmann-Doose K, Wittig T, Kraft K. Virucidal and virostatic in vitro activity of ELOM-080 against respiratory pathogens. Phytomedicine Plus 2021; 1: 100035
    CrossrefPubMedGoogle Scholar
  • 16 da Silva JKR, Figueiredo PLB, Byler KG, Setzer WN. Essential oils as antiviral agents, potential of essential oils to treat SARS-CoV-2 infection: an in-silico investigation. Int J Mol Sci 2020; 21: 3426
    Google Scholar
  • 17 Ma L, Yao L. Antiviral effects of plant-derived essential oils and their components: an updated review. Molecules 2020; 25: 2627
    CrossrefPubMedGoogle Scholar
  • 18 Wani AR, Yadav K, Khursheed A, Rather MA. An updated and comprehensive review of the antiviral potential of essential oils and their chemical constituents with special focus on their mechanism of action against various influenza and coronaviruses. Microb Pathog 2021; 152: 104620
    CrossrefPubMedGoogle Scholar
  • 19 Asif M, Saleem M, Saadullah M, Yaseen HS, Al Zarzour R. COVID-19 and therapy with essential oils having antiviral, anti-inflammatory, and immunomodulatory properties. Inflammopharmacology 2020; 28: 1153-1161
    Google Scholar
  • 20 Nadjib BM. Effective antiviral activity of essential oils and their characteristic terpenes against coronaviruses: an update. J Pharmacol Clin Toxicol 2020; 8: 1138
    PubMedGoogle Scholar
  • 21 Ojah EO. Exploring essential oils as prospective therapy against the ravaging coronavirus (SARS-CoV-2). Iberoam J Med 2020; 02: 322-330
    CrossrefPubMedGoogle Scholar
  • 22 Patne T, Mahore J, Tokmurke P. Inhalation of essential oils: could be adjuvant therapeutic strategy for COVID-19. Int J Pharm Sci Res 2020; 11: 4095-4103
    PubMedGoogle Scholar
  • 23 Li X, He X, Chen S, Guo X, Bryant MS, Guo L, Manjanatha MG, Zhou T, Witt KL, Mei N. Evaluation of pyrrolizidine alkaloid-induced genotoxicity using metabolically competent TK6 cell lines. Food Chem Toxicol 2020; 145: 111662
    CrossrefPubMedGoogle Scholar
  • 24 Roeder E. Medicinal plants in China containing pyrrolizidine alkaloids. Pharmazie 2000; 55: 711-726
    Google Scholar
  • 25 Kempf M, Schreier P, Reinhard A, Beuerle T. Pyrrolizidinalkaloide in Honig und Pollen. J Verbr Lebensm 2010; 5: 393-406
    CrossrefPubMedGoogle Scholar
  • 26 Wiedenfeld H, Edgar J. Toxicity of pyrrolizidine alkaloids to humans and ruminants. Phytochem Rev 2011; 10: 137-151
    CrossrefPubMedGoogle Scholar
  • 27 Merz KH, Schrenk D. Interim relative potency factors for the toxicological risk assessment of pyrrolizidine alkaloids in food and herbal medicines. Toxicol Lett 2016; 263: 44-57
    CrossrefPubMedGoogle Scholar
  • 28 Gao L, Rutz L, Schrenk D. Structure-dependent hepato-cytotoxic potencies of selected pyrrolizidine alkaloids in primary rat hepatocyte culture. Food Chem Toxicol 2020; 135: 110923
    CrossrefPubMedGoogle Scholar
  • 29 Rutz L, Gao L, Küpper JH, Schrenk D. Structure-dependent genotoxic potencies of selected pyrrolizidine alkaloids in metabolically competent HepG2 cells. Arch Toxicol 2020; 94: 4159-4172
    CrossrefPubMedGoogle Scholar
  • 30 Glück J, Waizenegger J, Braeuning A, Hessel-Pras S. Pyrrolizidine alkaloids induce cell death in human HepaRG cells in a structure-dependent manner. Int J Mol Sci 2021; 22: 202
    CrossrefPubMedGoogle Scholar
  • 31 Zheng P, Xu Y, Zhenhui R, Wang Z, Wang S, Xiong J, Zhang H, Jiang H. Toxic prediction of pyrrolizidine alkaloids and structure-dependent induction of apoptosis in HepaRG cells. Oxid Med Cell Longev 2021; 2021: 8822304
    CrossrefPubMedGoogle Scholar
  • 32 Lester C, Troutman J, Obringer C, Wehmeyer K, Stoffolano P, Karb M, Xu Y, Roe A, Carr G, Blackburn K, Mahony C. Intrinsic relative potency of a series of pyrrolizidine alkaloids characterized by rate and extend of metabolism. Food Chem Toxicol 2019; 131: 110523
    CrossrefPubMedGoogle Scholar
  • 33 Ning J, Chen L, Strikwold M, Louisse J, Wesseling S, Rietjens IMCM. Use of an in vitro-in silico testing strategy to predict inter-species and inter-ethnic human differences in liver toxicity of the pyrrolizidine alkaloids lasiocarpine and riddelliine. Arch Toxicol 2019; 93: 801-818
    CrossrefPubMedGoogle Scholar
  • 34 Ning J, Chen L, Rietjens IMCM. Role of toxicokinetics and alternative testing strategies in pyrrolizidine alkaloid toxicity and risk assessment; state-of-the-art and future perspectives. Food Chem Toxicol 2019; 131: 110572
    CrossrefPubMedGoogle Scholar
  • 35 Enge AM, Kaltner F, Gottschalk C, Braeuning A, Hessel-Pras S. Active transport of hepatotoxic pyrrolizidine alkaloids in HepaRG cells. Int J Mol Sci 2021; 22: 3821
    CrossrefPubMedGoogle Scholar
  • 36 Bundesinstitut für Arzneimittel und Medizinprodukte (BfArM). Bekanntmachung zur Prüfung des Gehalts an Pyrrolizidinalkaloiden zur Sicherstellung der Qualität und Unbedenklichkeit von Arzneimitteln, die pflanzliche Stoffe bzw. pflanzliche Zubereitungen oder homöopathische Zubereitungen aus pflanzlichen Ausgangsstoffen als Wirkstoffe enthalten. Announcement from 01. March 2016. Accessed April 28, 2021 at: http://www.bfarm.de/SharedDocs/Bekanntmachungen/DE/Arzneimittel/besTherap/bm-besTherap-20160301-pa-pdf.pdf
    Google Scholar
  • 37 Nowak M, Wittke C, Lederer I, Klier B, Kleinwächter M, Selmar D. Interspecific transfer of pyrrolizidine alkaloids: an unconsidered source of contaminations of phytopharmaceuticals and plant derived commodities. Food Chem 2016; 213: 163-168
    CrossrefPubMedGoogle Scholar
  • 38 Selmar D, Wittke C, Beck-von Wolffersdorff I, Klier B, Lewerenz L, Kleinwächter M, Nowak M. Transfer of pyrrolizidine alkaloids between living plants: a disregarded source of contamination. Environ Pollut 2019; 248: 456-461
    CrossrefPubMedGoogle Scholar
  • 39 Wiesner J, Reh K, Knöss W. Regulatorische Aspekte zu PA-Kontaminationen in Arzneipflanzen (Regulatory aspects concerning PA contamination in medicinal plants). J Kulturpflanzen 2020; 72: 84-87
    PubMedGoogle Scholar
  • 40 Steinhoff B. Pyrrolizidine alkaloid contamination in herbal medicinal products: limits and occurrence. Food Chem Toxicol 2019; 130: 262-266
    CrossrefPubMedGoogle Scholar
  • 41 Bundesinstitut für Risikobewertung (BfR). Bestimmung von Pyrrolizidinalkaloiden (PA) in Pflanzenmaterial mittels SPE-LC-MS/MS. Method description BfR-PA-Tee-2.0/2014. Accessed April 28, 2021 at: http://www.bfr.bund.de/cm/343/bestimmung-von-pyrrolizidinalkaloiden.pdf
    Google Scholar
  • 42 Bundesverband der Arzneimittel-Hersteller (BAH) & Bundesverband der Pharmazeutischen Industrie (BPI). Mögliche Kontaminationen mit Pyrrolizidinalkaloiden: Anforderungen an die produktspezifische Dokumentation der Qualitätskontrolle. Recommendations of the associations BAH and BPI from 16. December 2016. Accessed April 28, 2021 at: BAH-Homepage; BPI-Homepage: membernet.bpi.de/bibliothek/alle-downloads/8883.download?cHash=3742927fe8665;b2fea606f7d9999e6d5: http://www.bah-bonn.de/themen/meldung/pyrrolizidinalkaloide-empfehlungen-fuer-die-firmen-zur-umsetzung-der-bfarm-bekanntmachung-vom-1-maerz-2016
    Google Scholar
  • 43 Steinhoff B. Pflanzliche Arzneimittel: Aktuelle Anforderungen an die Prüfung auf Kontaminationen (Herbal medicinal products: current requirements for testing for contamination). Z Phytother 2017; 38: 166-169
    Article in Thieme ConnectPubMedGoogle Scholar
  • 44 Rassem HHA, Nour AH, Yunus RM. Techniques for extraction of essential oils from plants: a review. Aust J Basic Appl Sci 2016; 10: 117-127
    PubMedGoogle Scholar
  • 45 Burger P, Plainfossé H, Brochet X, Chemat F, Fernandez X. Extraction of natural fragrance ingredients: history overview and future trends. Chem Biodivers 2019; 16: e1900424
    CrossrefPubMedGoogle Scholar
  • 46 Sadgrove N, Jones G. A contemporary introduction to essential oils: chemistry, bioactivity and prospects for Australian agriculture. Agriculture 2015; 5: 48-102
    CrossrefPubMedGoogle Scholar
  • 47 Committee on Herbal Medicinal Products (HMPC). Guideline on good agricultural and collection practice (GACP) for starting materials of herbal origin. EMEA/HMPC/246816/2005 from 20. February 2006. Accessed April 28, 2021 at: http://www.ema.europa.eu/en/documents/scientific-guideline/guideline-good-agricultural-collection-practice-gacp-starting-materials-herbal-origin_en.pdf
    Google Scholar
  • 48 Kempf M, Heil S, Haßlauer I, Schmidt L, von der Ohe K, Theuring C, Reinhard A, Schreier P, Beuerle T. Pyrrolizidine alkaloids in pollen and pollen products. Mol Nutr Food Res 2010; 54: 292-300
    CrossrefPubMedGoogle Scholar
  • 49 Kempf M, Reinhard A, Beuerle T. Pyrrolizidine alkaloids (PAs) in honey and pollen – legal regulation of PA levels in food and animal feed required. Mol Nutr Food Res 2010; 54: 158-168
    CrossrefPubMedGoogle Scholar
  • 50 Committee on Herbal Medicinal Products (HMPC). Public statement on contamination of herbal medicinal products/traditional herbal medicinal products with pyrrolizidine alkaloids. EMA/HPMC/328782/2016 from 31 May 2016. Accessed April 28, 2021 at: http://www.ema.europa.eu/docs/en_GB/document_library/Public_statement/2016/06/WC500208195.pdf
    Google Scholar
  • 51 U.S. Environmental Protection Agency (EPA). Predicted average values of octanol-water partition coefficients (log P) of retrorsine (0.150), retrorsine N-oxide (− 1.01), lycopsamine (0.190) and limonene (4.55) can be found here: Accessed April 28, 2021 at: https://comptox.epa.gov/dashboard/dsstoxdb/results?search=retrorsine, https://comptox.epa.gov/dashboard/dsstoxdb/results?search=retrorsine N-oxide, https://comptox.epa.gov/dashboard/dsstoxdb/results?search=lycopsamine, https://comptox.epa.gov/dashboard/dsstoxdb/results?search=limonene.
    Google Scholar
  • 52 Committee on Herbal Medicinal Products (HMPC). Public statement on the use of herbal medicinal products containing toxic, unsaturated pyrrolizidine alkaloids (PAs) including recommendations regarding contamination of herbal medicinal products with pyrrolizidine alkaloids. EMA/HPMC/893108/2011 Rev. 1 (draft) from 08. July 2020. Accessed April 28, 2021 at: http://www.ema.europa.eu/en/documents/public-statement/draft-public-statement-use-herbal-medicinal-products-containing-toxic-unsaturated-pyrrolizidine_en-0.pdf
    Google Scholar
  • 53 European Medicines Agency (EMEA). ICH Topic Q2 (R1). Note for guidance on validation of analytical procedures: text and methodology. Step 5 (CPMP/ICH/381/95) June 1995. Accessed April 28, 2021 at: http://www.ema.europa.eu/en/documents/scientific-guideline/ich-q-2-r1-validation-analytical-procedures-text-methodology-step-5_en.pdf
    Google Scholar
  • 54 [Anonymous] PA/PH/Exp. PA/T (18) 1 ANP, 2.8.26. Contaminant pyrrolizidine alkaloids. Pharmeuropa 32.1 (January 2020). Accessed April 28, 2021 at: http://pharmeuropa.edqm.eu/app/Archives/content/Archives-0/Pharmeuropa3201E.pdf
    Google Scholar
  • 55 [Anonymous] Pesticide residues. European Directorate for the Quality of Medicines & HealthCare. European Pharmacopoeia, 10th Edition (Supplement 10.5). General Chapter 2.8.13. Accessed April 28, 2021 at: https://www.edqm.eu/en
    Google Scholar
  • 56 [Anonymous] European Commission. Analytical quality control and method validation procedures for pesticide residues analysis in food and feed. SANTE/12682/2019, 01/01/2020. Accessed April 28, 2021 at: http://www.eurl-pesticides.eu/userfiles/file/EurlALL/AqcGuidance_SANTE_2019_12682.pdf
    Google Scholar