Contamination of Teas and Herbal Infusions – a Current Review of Findings for Pyrrolizidine Alkaloids and Further Contaminants

Contamination of teas and herbal infusions – a current review of findings for pyrrolizidine alkaloids and further contaminants

Eurofins Dr. Specht Laboratorien GmbH Johannes Jaschik CoTeCa 2018

www.eurofins.de Contaminants ABC

Anthraquinone Nicotine Biphenyl, Bromide Ortho-Phenylphenol (2-Phenylphenol) Chlorate, Copper Pyrrolizidine alkaloids, Perchlorate, PAHs … Diethyltoluamid (DEET) QACs… Ethoxyquin Rare earth elements Folpet, Fluoride Smoke, S421 G… Trimethylsulfonium, Tallow amines Hexachlorobenzene U… I… V… J… W… K… X… Lead Y… Mercury Z…

Johannes Jaschik, COTECA 2018 2 Residues vs. Contaminants

Residues: Contamination: Whatever remains after the Presence of an unwanted use of use or application of a chemical which not originate of certain chemical an use in the material.

from analytical perspective there is no difference

A residue may act as a contaminant in material or environment it was not intented to be used in.

Johannes Jaschik, COTECA 2018 3 Residues vs. Contaminants

Why is it important to differ between residues and contaminants:

• finding the source of the residue/contamination

• residues: checking the application (limit/change use)

• contaminants: checking environment (unavoidable?) • distribution of the residue/contamination

• often spot contamination, not equally distributed

• residues are more equally distributed • maybe different legal requirements

• e.g. organic production

Johannes Jaschik, COTECA 2018 4 Analytical options

How do you approach the assessment of contamination in tea and herbal infusion: Looking at your product…

There are as many different substances which act as contaminants as products available.

Johannes Jaschik, COTECA 2018 5 Examples for Categories

Mineral oils Photo initiators Heavy metals (MOSH/MOAH) (illegal) dyes Dioxins Radioactivity Allergenes Plasticizers Desinfectans Pesticides PAHs (e.g. QACs) GMO Irradiation Chlorate, Microbiology Perchlorate, Phosphoric acid, etc. Vet drugs … Fluoride … and other products in your portfolio

Johannes Jaschik, COTECA 2018 6 Offer with different methods

sample tea There is no „C.S.I. – analysis“ but quite a lot of specific work! all contaminants known What Eurofins can offer:

group specific Specific methods Multi methods e.g. heavy metals e.g. mycotoxins Method Multi pesticides Method many different methods, Specific methods vet drugs modern equipment, e.g. microbiology experience in evaluation

Johannes Jaschik, COTECA 2018 7 Keeping your risk low

Not to analyse everything but to control for your risks and specific checking these.

In order to have time and opportunity to…

Enjoy your product !

Johannes Jaschik, COTECA 2018 8 Example Lapsang souchong

Lapsang souchong […], sometimes referred to as smoked tea [..] is a black tea (Camellia sinensis) originally from the mountainous Wuyi region in the Chinese province of Fujian. It is distinct from other types of tea, as the leaves are traditionally smoke-dried over pinewood fires, imparting a distinctive flavor of smoky pine. (Wikipedia, 2018)

THIE Compendium of Guidelines for Tea, 2018

Johannes Jaschik, COTECA 2018 9 Contamination of Lapsang

Phthalimid Anthraquinone Biphenyl Cypermethrin Anthrachinon 2-Phenylphenol Bifenthrin Phthalimid (PI) Imidacloprid Bifenthrin Chlorfenapyr different ranking but Cypermethrin Thiamethoxam more important also Thiamethoxam Acetamiprid different concentrations Cyhalothrin, lambda- Cyhalothrin, lambda- Acetamiprid Thiacloprid Thiacloprid Carbendazim Glyphosate

ranking for ranking for all teas Lapsang

Johannes Jaschik, COTECA 2018 10 Lapsang contaminants

Teas except Lapsang Lapsang 90,0 90,0 80,0 80,0 70,0 70,0 60,0 60,0 50,0 50,0 Anthrachinon 40,0 40,0 2-Phenylphenol 30,0 30,0 Phthalimid (PI) 20,0 20,0 Biphenyl 10,0 10,0 0,0 0,0

• completely different concentration • but each substance has slightly different distribution • keeping in mind, that Biphenyl/2-Phenyphenol are not common on Tea except Lapsang at all

Johannes Jaschik, COTECA 2018 11 Biphenyl (1/2)

Further contaminant from smoke and other sources. In the past used in citrus fruits. Detection in foodstuff with GC-MS using multi method possible.

Products MRLs Dried chilli 0.01 Dried fruits 0.01 Herbal infusions 0.05 Herbs 0.1 Mate 0.5 Nutmeg/Macis 1 Rice 0.01 Spices 0.05 Tea 0.05

Johannes Jaschik, COTECA 2018 12 Biphenyl (2/2)

Biphenyl: overview distribution 70,0

60,0

50,0

Evaluation for organic 40,0 conventional products takes into account 30,0 organic the different contamination 20,0 sources of Biphenyl. 10,0 But if the MRL is exceeded – 0,0 there is no further tolerance. <=0,01<=0,02<=0,05 <=0,1 <=0,2 <=0,5 >0,5

Graph for conventional and organic products comparable – contamination probable. Higher concentrations in mate and nutmeg.

Johannes Jaschik, COTECA 2018 13 Anthraquinone (1/3)

pesticide

Detection in foodstuff with GC-MS/MS using multi method possible.

Johannes Jaschik, COTECA 2018 14 Anthraquinone (2/3)

Products:

Products MRLs* Tea 0.02 herbal infusions 0.02 Packaging / inks Mate 0.02 packaging material Inks Herbs 0.01 Spices 0.02 dried fruits/vegetables 0.01 Anthraquinone: overview distribution *without concentration factor 60,0

50,0 Graph for conventional and 40,0 30,0 Conventional Organic organic products 20,0 comparable – 10,0 0,0 contamination probable. <=0,01 <=0,02 <=0,05 <=0,1 >0,1

Johannes Jaschik, COTECA 2018 15 Anthraquinone – source (3/3)

Smoke

Johannes Jaschik, COTECA 2018 16 2-Phenylphenol (1/2)

Detection in foodstuff with GC-MS 2-Phenolphenol: overview using multi method possible. distribution 35,0 30,0 25,0 20,0 conventional 15,0 organic More than one effect. 10,0 5,0 0,0 <=0,01<=0,02<=0,05 <=0,1 <=0,2 <=0,5 >0,5

Johannes Jaschik, COTECA 2018 17 2-Phenylphenol (2/2)

2-Phenylphenol: overview (without citrus) 50,0 Products MRLs 45,0 Citrus /oil 5 40,0 Coffee 0,1 35,0 30,0 Fruits 0,05 25,0 conventional Herbal infusions 0,1 20,0 organic 15,0 Ink / packaging 10,0 Spices 0,1 5,0 Tea 0,1 0,0 <=0,01<=0,02<=0,05 <=0,1 <=0,2 <=0,5 >0,5

Now the graph shows the typical curve for contamination Contamination source

Either use, as in citrus fruits, or Approved use contamination from packaging/ink or environment (smoke).

Johannes Jaschik, COTECA 2018 18 Folpet/Phthalimide

Phthalimide

Molar Mass: 147,13 g/mol

Phthalimide residues have Folpet to be multiplied by 2 in order to gain the Folpet residue.

Regulation (EC) No 396/2005 as it stood before being amended by this Regulation shall continue to apply to Molar Mass: 296,56 g/mol products which were produced before 26 August 2016.

Johannes Jaschik, COTECA 2018 19 Folpet/Phthalimide Relana/BNN

forming under heating conditions

Considering all arguments including the above mentioned, a positive finding of Phthalimide only is in our oppinion not sufficient to decide if regulation (EC) 396/2005 should be applied in this case.

Johannes Jaschik, COTECA 2018 20 Folpet/Phthalimide data review

Phthalimide: overview distribution Although some use of Folpet 45,0 (or Captan which contains 40,0 35,0 Folpet additionally) is known, 30,0 25,0 conventional contamination is the 20,0 prevalent factor of 15,0 organic 10,0 Phthalimide findings. 5,0 0,0 <=0,01<=0,02<=0,05 <=0,1 <=0,2 <=0,5 >0,5

Products MRLs If Folpet is actively used you can expect Tea 0,1 Phthalimide in neighbouring fields. Spices 0,1 Apple 0,03 Analysis possible with GC/MS(/MS) using multi Dried Chillies 0,03 methods for pesticides but use of isotope Herbal infusions 0,1 marked standards for Folpet and Phthalimide other fruits/concentrates 0,03 - 6 necessary.

Johannes Jaschik, COTECA 2018 21 Heavy metals

“…and some green tea leaves, particularly those from China, are contaminated with lead…”

https://well.blogs.nytimes.com/2013/05/23/whats-in-your-green-tea/

Plants can take up the heavy metals e.g. lead from the soil and under certain conditions high levels can be accumulated in the leaves and other edible parts of the plant. For tea plants it is known, that they can uptake heavy metals from the soils and a proportion will be transported to the tea leaves which are used for the tea infusion.

https://www.researchgate.net/publication/295858621_Metals_Contents_in_Black_Tea_and_Evaluation_of_Potential_Human_Health_Risks_to_Consumers Typically detected by single element analysis with AAS or ICP/MS.

Johannes Jaschik, COTECA 2018 22 Lead

Lead 40,0

35,0

30,0

25,0

20,0 conventional 15,0 organic 10,0

5,0

0,0 tea tobacco packaging material herbal infusions Only few data for organic available. spices

No MRLs for tea and herbal infusions in general, evaluation is only possible for selected commodoties according to regulation 1881/2006 .

Johannes Jaschik, COTECA 2018 23 Copper (1/2)

Plants can take up the heavy metals from the soil and under certain conditions high levels can be accumulated in leaves and other edible parts of the plant. Typically detected by single element analysis with AAS or ICP/MS.

Products MRLs Cocoa 50 Coffee 50 Herbal infusions 100 Herbs 20 Oilseeds 30-40 Products with copper Spices 40 Tea 40 concentrations exceeding 10mg/kg Tobacco

Johannes Jaschik, COTECA 2018 24 Copper (2/2)

Copper: overview distribution 70,0 Contamination? Use? 60,0 Depending on the product. 50,0 40,0

Use of copper is allowed in 30,0 conventional organic organic farming. 20,0

10,0

0,0 <=0,1 <=0,5 <=2 <=10 <=20 <=40 >40

Evaluation is done accordingly to EU regulation 396/2005.

Johannes Jaschik, COTECA 2018 25 Mercury (1/2)

Produkte MRLs Tea 0,02 Shrimps 0,5 Herbs 0,01* Herbal infusions 0,02 Bigger topic in seafood! Spices 0,02 Mushrooms 0,01 Tobacco

Johannes Jaschik, COTECA 2018 26 Mercury (2/2)

Mercury: overview distribution 60,0

50,0

40,0

Not surprisingly the 30,0 conventional graph shows a sign for 20,0 organic contamination. 10,0 0,0

Evaluation for products is done only according regulation 396/2005 with exception of

seafood where the evaluation is done according to contaminant regulation 1881/2006!

Johannes Jaschik, COTECA 2018 27 Chlorate / Perchlorate

EPRW 2016

BfR 2018

EFSA 2015

Johannes Jaschik, COTECA 2018 28 Chlorate (1/3)

• Defined as pesticide

• current toxicological evaluation (PRIMO)  high contents are being tolerated

• Draft document for new maximum residue levels SANTE 10684_2015  drastic lowering compared to PRIMO-model Detection in foodstuff with LC-MS/MS using a SANTE MRLs single method including Products [mg/kg] perchlorate. Milk 0.2 Starch (cereals) 0.04 Tea 0.15 Vegetables 0.01 - 0.7 Herbicide effects: destroys oxidative plant Fruits 0.015 - 0.03 tissue Herbal infusions 0.15 Typical byproduct of disinfectants, e.g. used for Herbs 0.6 chlorination of drinking water Coffee 0.15 …

Johannes Jaschik, COTECA 2018 29 Chlorate (2/3)

This would lead to serious problems with a wide range of foodstuff.

Johannes Jaschik, COTECA 2018 30 Chlorate (3/3)

Evaluation only according to PriMo regardless of cultivations status. Chlorate: overview distribution 35,0

30,0 This residue of Chlorate could be derived from a use of chlorinated resources during 25,0 20,0 production or processing or from the use of conventional chlorinated water due to disinfection 15,0 organic purposes. In many countries, a use of 10,0 chlorinated water is legal and partly 5,0

statutory to minimize microbiological risks. 0,0 […] <=0,01<=0,02<=0,05 <=0,1 <=0,2 <=0,5 >0,5 Based on the current EFSA model PRIMo, the exhaustion of ARfD of 0.036 mg/kg Several effects at work. bodyweight in the analyzed sample Chlorate is in use and this amounts x %. Therefore, an exceedance of the ARfD-value is not given. differently in conventional and organic production and Nevertheless, we recommend finding the in different products. source of entry and minimize levels of Chlorate in the product.

Johannes Jaschik, COTECA 2018 31 Perchlorate (1/3)

Exception because it is regulated as contaminant (not defined as pesticide!). However often monitored together with Chlorate.

Detection in foodstuff with LC-MS/MS using a single method including chlorate. Mainly anthropogenic origin - can occur naturally. Fireworks is suggested to be one source in specific origins.

Johannes Jaschik, COTECA 2018 32 Perchlorate (2/3)

Not regulated in the Perchlorate: overview distribution pesticide regulation. 70,0 60,0 50,0 40,0 30,0 conventional 20,0 organic 10,0 0,0

Contamination in food but non-food applications/use which lead to punctual high concentrations.

Johannes Jaschik, COTECA 2018 33 Perchlorate (3/3)

There is no specific conclusion for organic products. Perchlorate is treated in every respect as an contaminant.

Fruits Vegetables Herbs Perchlorate findings are Cereals sometimes specific for Tea Herbal Infusions certain origins e.g. China. Coffee Milk Spices

Johannes Jaschik, COTECA 2018 34 Pyrrolizidine alkaloids (PAs)

. PAs are produced by more than 6000 flowering plants (protection against herbivores)

. Most relevant plant is ragwort (Senecio jacobaea L.)

. 660 PAs and corresponding N-oxides are known; some of them being extremely toxic

. not clear scope: . BfR17 – BfR21 – BfR28 – or 35/38 or EFSA17 ????

Johannes Jaschik, COTECA 2018 35 Structures Pyrrolizidine alkaloids (PAs)

Retronicine

Lycopsamine Type Monocrotaline Type

Senecionine Type

Lycopsamine, Monocrotalin, Intermedine, Jacobine, Echimidine, Trichodesmine Lasiocarpine etc. Senecionine, Senkirkin, Retrorsine, etc.

Johannes Jaschik, COTECA 2018 36 Background Pyrrolizidine alkaloids (PAs)

. Unexpectedly high concentrations of PAs were found in (herbal) tea (0- 3.4 mg/kg dry product) (BfR statement 018/2013)

. 91 % of (herbal) teas contained PAs, 60 % of nutritional supplements; animal derived products only occasionally contaminanated (EFSA study, 2015, analysis of 1.000 samples)

. Affected products are:

tea (camellia sinensis) herbs, herbal teas honey, honey products salad, salad mixtures flour eggs, meat, milk (by carry-over)

Johannes Jaschik, COTECA 2018 37 PA Analysis @ Eurofins

Relevance . Further to the above mentioned matrices: pseudocereals, feed, borage oil, nutritional supplements, esp. products for infants, pregnant and nursing women . Continuous consumption of products containing high concentrations of PA is critical for infants, toddlers, pregnant and nursing women (BfR)

Specht Method . Existing LC-MS/MS method for BfR 32 PAs / N-oxides . Important: also for GMP requirements

Johannes Jaschik, COTECA 2018 38 Analytical view on PA

Retrorsine: negative in black tea

Seneciphylline N-oxide: added in black tea

Johannes Jaschik, COTECA 2018 39 Findings PA tea@ Eurofins

Retrorsine-NOx • Total number of samples: >4500 Lycopsamin-NOx Retrorsine (since Jan 2015) Senecionine-NOx • Results from zero to 10000 µg/kg in Jacobine-NOx Seneciphylline-NOx black tea Senecionine • Main PAs found (in all commodities): Indicine-NOx + Intermedine-NOx Indicine + Lycopsamine Jacobine Heliotrine-NOx Intermedine Senecivernine-NOx Lasiocarpin-NOx Senkirkine Europin-NOx Heliotrine Seneciphylline Europine N-oxide Lasiocarpine Europin Echimidine Senecivernine Echimidine-NOx

Johannes Jaschik, COTECA 2018 40 Review findings

Europine N-oxide Overview about PAs found Senecionine-NOx at least 10% in positive Heliotrine-NOx Lasiocarpin-NOx findings in 2018. Lycopsamin-NOx Retrorsine-NOx Senecionine Tea Seneciphylline-NOx Senecivernine-NOx Lycopsamin-NOx Retrorsine Jacobine-NOx Senkirkine Retrorsine-NOx Intermedine-N-Oxide / Indicine-N-Oxide Retrorsine Heliotrine

Jacobine Europin Intermedine Senecionine-NOx Senecivernine Echimidine-NOx More different PAs in herbal infusions. Herbal infusions

Johannes Jaschik, COTECA 2018 41 PAs in herbal infusions and tea

Sum of Pyrrolizidine alkaloids 50000 1000

45000 900

40000 800

35000 700 values values 30000 600 Maximum value (HI) max

average Maximum value (tea) 25000 500 Average values (HI) 20000 400 Average values (tea) 15000 300 concentration 10000 200 concentration

5000 100

0 0 2015 2016 2017 2018

No Monitoring data. Different effects: certain critical origins.

Johannes Jaschik, COTECA 2018 42 PAs in herbal infusions and tea

Average values (tea) 1600 Retrorsine-NOx 1400

High findings in 1200 Senecionine-NOx 2017 strongly 1000 800 Retrorsine connected to 600 Retrosine-NOx. concentration 400 sum of indicine + lycopsamine 200 sum of indicine-NOx + 0 intermedine-NOx 2015 2016 2017 2018

Average values (herbal infusions) 600

500 Composition of Retrorsine-NOx 400 Seneciphylline-NOx PAs depending 300 Senecionine-NOx Retrorsine on analysed 200 concentration Senecionine products. 100 Heliotrine-NOx 0 2015 2016 2017 2018

Johannes Jaschik, COTECA 2018 43 Tropane alkaloids (TAs) in the spotlight

. TA are secondary metabolic products formed to protect against herbivores by a great many of plant species . Well known plants are thorn apple (Datura stramonium), deadly nightshade (Atropa belladonna), henbane (Hyoscyamus niger) . More than 200 Substanzen are known; TAs investigated in detail are atropine ((-)- and (+)- hyoscyamine), scopolamine and cocaine

Toxikology: BfR-Statement 035/2014 and EFSA Statement 2013 . Group ARfD of 0.016 µg/kg might be exceeded by toddlers consuming cereals and cereal based products.

Johannes Jaschik, COTECA 2018 50

Structures Tropane alkaloids (TAs)

Tropin

Atropine, R- and S- Scopolamin (Hyoscyamin) (Hyoscin)

Further: Littorine, Norscopolamine, Homatropine, etc.

Johannes Jaschik, COTECA 2018 51 Findings TA tea@ Eurofins

• Total number of tests: >1800 (since Jan 2015) • Results from zero to > 9000 µg/kg in plant extracts • In tea up to 15 µg/kg (Atropin) • max findings in herbal infusion/raw material 2018

Atropine 9230 µg/kg Scopolamine 4360 µg/kg Atropine-NOx 31,8 µg/kg Anisodamine 290 µg/kg Norscopolamine 46 µg/kg

Johannes Jaschik, COTECA 2018 53 Contaminants in tea/herbal infusions

• contaminants are not caused by intended use of chemicals

• source can not determinated by analysis

• contamination can be reduced if source is known, but sometimes there are multiple sources for the same contaminant

• is not always avoidable (especially from soil, air or water)

• sometimes specific for certain origins

• there are always some more…

Johannes Jaschik, COTECA 2018 54 Thank you very much!

Contact

Johannes Jaschik: Fon: +49-(0)40-881 448-452 ? Fax: +49-(0)40-881 448-103 [email protected] www.eurofins.de

Johannes Jaschik, COTECA 2018 55