Determination of pyrrolizidine alkoloids in oregano (Origanum onites) produced in Denizli by LC-MS Q-TOF method

Yıl 2022, , 115 - 123, 15.06.2022

Seda Dicle Korkmaz , Özlem Küplülü

https://doi.org/10.33188/vetheder.1037994

Cited By: 1

Öz

Pyrrolizidine alkaloids (PAs) in certain plant species create contamination, especially in production areas, and pose a risk to public health with the consumption of foods. Due to their sensitivity to intoxications, it has hepatotoxic, pneumotoxic, genotoxic, mutagenic, teratogenic and carcinogenic effects in all age groups, especially in infants and children. In this study, the presence and level of PAs and N-oxide compounds in oregano grown in Denizli, which has an important share in Turkey's oregano export, were investigated by LC-MS Q-TOF to determine potential public health risks. In the study, a total of 3 processed and packaged oregano samples taken from different enterprises producing oregano in Denizli and 8 unprocessed (stem) oregano samples from different regions of Denizli (11 samples ) were used as material. Identification and quantification of PAs and PANOs compounds in the samples were done by LC-MS Q-TOF. A total of 280 ±22 µg/kg of europine and europine N-oxide were determined in a processed packaged sample from the analyzed samples. According to the analysis results; PA value in oregano samples was determined below the PA limit value (<1000 µg/kg) specified in the regulation no 2020/2040/EU. It was determined that the samples did not pose a risk to public health in terms of PAs. However, the fact that no PAs was found in the processed and packaged sample in which europin was detected and in the intact oregano samples taken from the same region indicates PAs contamination caused by weed and seed pieces, albeit in trace amounts, during the processing of oregano. It is reported that the PAs limit value for plant-derived foods are specified in the Commission Regulation No 2020/2040/EU, and this value going to be 1000 µg/kg for oregano with the regulations to be made in 2022. It is thought that this study will lay the ground work for PAs regulations within the scope of the Turkish Food Codex.

Anahtar Kelimeler

Denizli, LC-MS Q-TOF, Oregano, Pyrrolizidine alkaloids

Denizli ilinde üretilen kekiklerde (Origanum onites) pirolizidin alkaloidlerinin LC-MS Q-TOF yöntemi ile belirlenmesi

Öz

Belirli bitki türlerinde bulunan Pirolizidin alkaloidleri (PA), özellikle üretim alanlarında kontaminasyon oluşturarak gıdaların tüketimiyle halk sağlığı açısından risk oluşturmaktadır. PA'lar, intoksikasyonlara duyarlılıklarından dolayı başta bebek ve çocuklar olmak üzere tüm yaş gruplarında hepatoksik, pnömotoksik, genotoksik, mutojenik, teratojenik ve karsinojenik etkilidir. Çalışmada, Türkiye’nin kekik ihracaatında önemli payı olan Denizli’de yetiştirilen kekiklerde (Origanum onites) PA ve N-oksit (PANO) bileşiklerinin varlığının ve düzeyinin LC-MS Q-TOF ile araştırılarak potansiyel halk sağlığı risklerinin belirlenmesi amaçlandı. Denizli’de kekik üretimi yapan farklı işletmelerden alınan işlenmiş, paketlenmiş 3 kekik örneği ile Denizli ilinin farklı bölgelerine ait işlenmemiş 8 kekik örneği olmak üzere toplam 11 örnek materyal olarak kullanıldı. Örneklerde PA ve PANO bileşiklerinin tanımlanması ve miktarlandırılması LC-MS Q-TOF ile yapıldı. Analize alınan örneklerden işlenmiş bir örnekte, europine ve europine N-oksit toplam 280 ±22 µg/kg tayin edildi. Analiz sonuçlarına göre; kekik örneklerindeki PA değeri, 2020/2040/AB sayılı regülasyonda belirtilen PA limit değerin (1000 µg/kg) altında saptandı. Örneklerin, PA yönüyle halk sağlığı açısından risk oluşturmadığı belirlendi. Ancak europin saptanan işlenmiş ve paketlenmiş örnek ile aynı bölgeden alınan bütünlüğü bozulmamış kekik örneklerinde PA'ya rastlanmamış olması kekiğin işlenme aşamasında iz miktarda da olsa yabani ot ve tohum parçalarından kaynaklanan kontaminasyonunu işaret etmektedir. Çalışma, alkaloid kontaminasyonunu önlemek amacıyla kekik üretiminde işlenme sürecinin önemini ve ürünün yabani otlardan arındırılması gerekliliğini ortaya koymuştur. Bitki kaynaklı gıdalar için PA sınır değerlerinin 2020/2040/AB sayılı Komisyon Tüzüğü’nde belirtildiği, 2022 yılında yapılacak düzenlemelerle kekik için bu değerin 1000µg/kg olacağı bildirilmektedir. Çalışmanın Türk Gıda Kodeksi kapsamında PA düzenlemelerine zemin oluşturacağı öngörülmektedir.

Anahtar Kelimeler

Denizli, Kekik, LC-MS Q-TOF, Pirolizidin alkaloidleri

Kaynakça

• BfR (Federal Institute for Risk Assessment) (2016): Opinion No. 030/2016: Pyrrolidine Alkaloids: Level in Food should Continue to be Kept as Low as Possible; BfR: Berlin, Germany, Available from: URL:https://www.bfr.bund.de/cm/349/pyrrolizidine-alkaloids-levels-in-foods-should-continue-to-be-kept-as-low-as-possible.pdf
• Boppre, M (2011): The ecological context of pyrrolizidine alkaloids in food, feed and forage: an overview. Food Addit Contam Part A, 28(3), 260-281. Chen L, Mulder PPJ, Louisse J, Peijnenburg A, Wesseling S, Rietjens IMCM (2017): Risk assesment for pyrrolizidine alkaloids detected in (herbal) teas and plant food supplements. Regul Toxicol Pharmacol, 86, 292-302.
• Chen Z, Huo JR (2010): Hepatic veno-occlusive disease associated with toxicity of pyrrolizidine alkoloids in herbal preparations. Neth J Med, 68(6), 252-260.
• Crews C, Berthiller F, Krska R (2010): Update on analytical methods for toxic pyrrolizidine alkaloids. Anal Bioanal Chem, 396 (1), 327-338.
• Dusemund B, Nowak N, Sommerfeld C, Lindtner O, Schäfer B, Lampen A (2018): Risk assessment of pyrrolizidine alkaloids in food of plant and animal origin. Food Chem Toxicol, 115, 63,72.
• Edgar JA, Colegate SM, Boppre M., Molyneux RJ (2011): Pyrrolizidine alkaloids in food: a spectrum of potential health consequences. Food Addit Contam Part A, 28(3), 308-324.
• European Food Safety Authority (2011). Scientific Opinion on Pyrrolizidine alkaloids in food and feed. EFSA J. 2011, 9, 1–134.
• European Union (2020). Commission Regulation (EU) 2020/2040 of 11 December 2020 amending Regulation (EC) No 1881/2006 as regards maximum levels of pyrrolizidine alkaloids in certain foodstuffs. Off J Eur Union L. 420:1–5. [accessed 2021 Jun 18]. Available from: URL:https://eur-lex.europa.eu/legal-content/EN/TXT/?uri=CELEX:32020R2040
• Fu PP, Xia Q, Lin G., Chou, MW (2004): Pyrrolizidine alkaloids-genotoxicity, metabolism enzymes, metabolic activation, and mechanisms. Drug Metabol Rev, 36(1), 1-55.
• Izcara S, Casado N, Zarcero SM, Sierra I (2020): A Miniaturized QuEChERS Method Combined with Ultrahigh Liquid Chromatography Coupled to Tandem Mass Spectrometry for the Analysis of Pyrrolizidine Alkaloids in Oregano Samples. Foods, 9(9), 1319.
• Kaltner F, Rychlik M, Gareis M, Gottschalk C (2018): Influence of Storage on the Stability of Toxic Pyrrolizidine Alkaloids and Their N-Oxides in Peppermint Tea, Hay, and Honey. J Agric Food Chem, 66(20), 5221-5228.
• Kaltner F, Rychlik M, Gareis M, Gottschalk C (2020): Occurrence and Risk Assessment of Pyrrolizidine Alkaloids in Spices and Culinary Herbs from Various Geographical Origins. Toxins, 12(3), 155.
• Karlı B, Demir Z, Dalgıç A (2020): Denizli ilinde kekik üretimi yapan işletmelerin sosyo-ekonomik yapısı ve sorunları. Isparta Uyg Bilim Ziraat Fak Derg, 15(2), 151, 160.
• Kempf M, Wittig M, Schönfeld K, Cramer L, Schreier P, Beuerle T (2011). Pyrrolizidine alkaloids in food: downstream contamination in the food chain caused by honey and pollen. Food Addit Contam Part A, 28(3), 325-331.
• Kempf M, Reinhard A,Beuerle T (2010): Pyrrolizidine alkaloids (PAs) in honey and pollen‐legal regulation of PA levels in food and animal feed required. Mol Nutr Food Res, 54(1), 158-168.
• Molyneux RJ, Gardner, DL, Colegate, SM, Edgar, JA (2011):Pyrrolizidine alkaloid toxicity in livestock: a paradigm for human poisoning. Food Addit Contam: Part A, 28(3), 293-307.
• Muca B, Yıldırım B, Özçelik Ş, Koca A (2012): Isparta’s (Turkey) pisonous plants of public access places. Biological Diversity and Conversation, 5(1), 23-30.
• Mudge, EM, Maxwell PJ, Paula NB (2015): Quantification of pyrrolizidine alkaloids in North American plants and honey by LC-MS:Single laboratory validation. Food Addit Contam Part A, 32(12), 2068,2074.
• Nemli Y, Kaynar A, Kayadan A, Er T, Kaya İ (2015): Cuscuta campestris’in Pyrrolizidine Alkaloid içeriğine ilişkin ilk kanıt. Turk J Weed Sci, 18(2), 23-25.
• Ozansoy G, Küplülü Ö (2017): Importance of Pyrrolizidine Alkaloids in Bee Products. Mellifera, 17(1), 1,8.
• Picron J-F, Herman M, Hoeck EV, Goscinny S (2018):Analytical strategies for the determination of pyrrolizidine alkaloids in plant based food and examination of the transfer rate during the infusion process. Food Chem, 266, 514,523.
• Scharmm S, Köhler N, Rozhon W (2019): Pyrrolizidine alkaloids: biosynthesis, biological activities and occurrence in crop plants. Molecules, 24, 498.
• Schrenk D, Gao L, Lin G, Mahony C, Mulder PPJ, Peijnenburg A, Pfuhler S, Rietjens IMCM, Rutz L, Steinhoff B, These A (2020): Pyrrolizidine alkaloids in food and phytomedicine: Occurence, exposure, toxicity, mechanisms and risk assesment- A review. Food Chem Toxicol, 136, 111107.
• Sokat Y (2020): Kekik üretim alanlarında görülen bazı zararlı yabancı ot türleri. Bahri Dağtaş Bitkisel Araştırma Dergisi, 9(1), 29-42.
• Than KA, Stevens V, Knill A, Gallagher PF, Gaul KL, Edgar JA (2005): Plant-associated toxins in animal feed: screening and confirmation assay development. Anim Feed Sci Tech, 121, 5-21.
• Tosun F, Tamer U (2004): Determination of Pas in the Seeds of Heliotropium Europaeum by GC/MS. J Fac Pharm Ankara, 33 (1), 7,9.
• TUİK (2021): Bitkisel Üretim İstatistikleri, Erişim: https://biruni.tuik.gov.tr/medas/?kn=92&locale =tr, Erişim tarihi:4.9.2021 29. Wiedenfeld H (2011): Toxicity of Pyrrolizidine Alkaloids–a Serious Health Problem. J Marmara Univ Inst Health Sci, 1(2), 79-87. Wiedenfeld, H, Edgar, J (2011). Toxicity of pyrrolizidine alkaloids to humans and ruminants. Phytochem Rev, 10(1), 137-151.