The laboratory carries out the determination of several groups of plant alkaloids and other biologically active substances in various plant and animal materials and foods. The specifications of each group of substances and the methods for their analysis are given below. The analytical methods used comply with the requirements of Implementing Regulation (EU) 2023/2783 laying down methods of sampling and analysis for the control of plant toxins in foodstuffs.
Tropane alkaloids
Tropane alkaloids are natural toxins produced by plants, for example, in the Solanaceae family, such as Datura stramonium, or in the Convolvulaceae family, such as Convolvulus arvensis. Their determination is performed by multidetection U-HPLC-MS/MS (KM 23A) or HRMS/MS, which includes up to 21 alkaloids, including atropine and scopolamine. Qualitative screening for the presence of other tropane alkaloids for which analytical standards are not available can also be performed using the U-HPLC-HRMS/MS method. Tropane alkaloids are mainly determined in foods of plant origin, cereals, pseudocereals, herbal teas, spices, food supplements based on herbal extracts, baby food, crops and feeds. Commission Regulation (EU) 2023/915 set maximum levels for atropine and scopolamine.
Pyrrolizidine and quinolizidine alkaloids
Pyrrolizidine alkaloids and their N-oxides belong to a wide group of natural toxins occurring in medicinal and weedy plants of many families such as Borageaceae, Asteraceae, Fabaceae, Rhamnaceae, Heliotropicae, and others. Quinolizidine alkaloids are naturally occurring toxic substances produced by plants of the legume family (Fabaceae), such as lupine or lupine in particular. The determination of these alkaloids is performed by multidetection U-HPLC-MS/MS (KM 23A) or HRMS/MS, which includes up to 33 pyrrolizidine and 2 quinolizidine alkaloids. Qualitative screening can also be performed using U-HPLC-HRMS/MS to detect the presence of additional alkaloids for which analytical standards are not available. Pyrrolizidine and quinolizidine alkaloids are determined in foods of plant origin, cereals, pseudocereals, herbal teas, spices, food supplements based on herbal extracts, baby food, crops, animal feed, as well as pollen and honey. Commission Regulation (EU) 2023/915 set maximum levels for the sum of all 21 pyrrolizidine alkaloids, including their N-oxides.
Opium alkaloids
Opium alkaloids are typical representatives of the natural toxins contained in the capsule of the poppy (Papaver somniferum). Their determination is performed by multidetection U-HPLC-HRMS (KM 23B), which includes up to 7 alkaloids, including morphine and codeine. Opium alkaloids are determined in poppy seeds and other oilseeds, in poppy seeds and in bakery products. The determination of opium alkaloids allows the quality and safety of the raw material used to be assessed. This allows us to detect poor purification of poppy seeds from poppy seed pulp, or even the possible substitution of food poppy seeds with technical poppy seeds, the consumption of which may be a health risk for consumers. The maximum limit set by Commission Regulation (EU) 2023/915 in products for final consumers applies to the sum of morphine + 0.2 x codeine. According to Decree 329/1997 Coll., poppy containing more than 25 mg/kg of morphine alkaloids (morphine, codeine and thebaine) may not be used for food purposes.
Glycoalkaloids
Glycoalkaloids are natural toxins which are found mainly in potatoes, a crop belonging to the aubergine family (Solanaceae). Their determination is carried out by the multidetection method U-HPLC-HRMS (KM 23C), which includes the glycoalkaloids alpha-solanine and alpha-chaconine but also their degradation products, which have the same toxicity. Glycoalkaloids are determined in potatoes and potato products. Commission Recommendation (EU) 2022/561 gives an indicative (maximum) value of 100 mg/kg for the sum of solanine and chaconine for these commodities.
Capsaicinoids and piperine
Capsaicinoids are among the alkaloids uniquely found in the fruits of the Capsicum or chilli plant and are responsible for their pungency. Piperine is the main pungent component of various peppers, especially black pepper (Piper nigrum). The determination of these compounds is carried out by the multidetection method U-HPLC-HRMS (KM 23E), which includes capsaicin, dihydrocapsaicin, nordihydrocapsaicin and N-vanillylnonanamide or piperine. The determination of capsaicinoids and piperine is carried out in spices, fresh and dried peppers, and chilli chips. The study of the chemical processes involved in the pungency of peppers can serve to improve the taste and quality of foods or flavourings containing them; of importance in this respect is not only information on the exact content of capsaicinoids, but also the determination of their abundance and distribution in different parts of the plant. Outside the scope of accreditation, we can also estimate the hotness of peppers (chilli) in Scoville units (SHU) or determine the content of extractable dyes in ASTA units (EN ISO 7541).
Alkaloids of Mitragyna sp. (kratom)
Indole alkaloids of Mitragyna speciosa are the main active substances of this tree belonging to the Rubiaceae family. Their determination is performed by a multidetection U-HPLC-HRMS/MS method (KM 20A), which includes 6 alkaloids including mitragynine, 7-hydroxymitragynine and mitraphylline and also allows qualitative screening of other alkaloids of this plant. The determination of these alkaloids is carried out in Mitragyna speciosa-based products, such as the typically dried leaf powder referred to as kratom. The analysis carried out may serve to check the quality and authenticity of the products and raw materials offered, or contribute to a better understanding of the relationship between the alkaloid content and the effects of kratom on the human body.
Alkaloids of mushrooms
Mushroom alkaloids are biologically active substances contained in certain genera of mushrooms, such as Psilocybe and Amanita. Their determination is carried out by the multidetection U-HPLC-HRMS method (KM 20C), which has two variants of design, namely for the separate determination of alkaloids of Psylocibe (psilocin and psilocybin) and for the separate determination of alkaloids of Amanita (muscimol, ibotenic acid and muscarinic acid). The above alkaloids are determined in fresh and dried mushrooms, extracts and tinctures. The determination of these alkaloids provides a deeper understanding of the composition and biological activity of these mushrooms. Psilocin and psilocybin are known for their psychotropic effects, reflecting their importance in ethnobotany and human behavioural research. On the other hand, Amanita alkaloids, particularly muscimol and ibotenic acid, have neurological effects and are being studied for their potential importance in the treatment of neurological diseases.
Coca alkaloids
Coca alkaloids are a group of tropane alkaloids found in plants of the genus Erythroxylon (coca bushes). Their determination is carried out by a multidetection U-HPLC-HRMS/MS method (KM 20B), which includes cocaine and ecgonine, and also allows qualitative screening of other alkaloids of this plant, such as anhydroecgonine and benzoylecgonine. Coca alkaloids are determined in solid and liquid samples of plant materials, extracts and products containing them.
Silymarin
Silymarin compounds are biologically active substances from the group of flavonolignans or flavonoids found in the extract of the plant Silybum marianum. The mixture of predominantly these substances is referred to as the silymarin or silymarin complex, the main component being a mixture of two diastereoisomers, silybin A and silybin B, collectively referred to as silibinin. Determination of the silymarin components is performed by multidetection U-HPLC-HRMS (KM 23D), which allows quantitative determination of silybin as well as qualitative screening (with the possibility of semiquantitative estimation) of up to 11 other components of this complex. Silymarin components are determined in milk thistle seeds and in various dietary supplements often intended to support proper liver function. The analysis performed allows the quality control of the raw material for the grower or processor, or verification of the declaration of silymarin content in dietary supplements.
Synephrine
Synephrine is a natural alkaloid that is one of the active ingredients in dietary supplements for weight loss, typically in the form of bitter orange (Citrus aurantium) extract. Its determination in dietary supplements is performed by U-HPLC-HRMS(MS) (KM 23F). Decree 58/2018 Coll. on food supplements and food composition sets out the conditions for the use of Citrus aurantium (or its extract converted to synephrine) in food supplements, so that the maximum permissible amount in a daily dose is 20 mg of synephrine.
Vanillin
Vanillin is a bioactive plant metabolite chemically classified in the group of phenols. It is the main aromatic component of the extract of vanilla pods, i.e. the fruits of plants of the vanilla genus (Vanilla; especially the species V. planifolia). The determination of vanillin in vanilla pods is carried out using the U-HPLC-HRMS(MS) method (KM 23G). It is possible to determine the water content in vanilla pods and to express the vanillin content in dry matter of the sample (out of the accreditation scope).