20.500.12556/RUL-177318 Ugotavljanje kakovosti prehranskih dopolnil s tetrahidrofolno kislino Evaluation of the quality of food supplements with tetrahydrofolic acid Folati oz. vitamin B9 so ključni za ohranjanje zdravja, njihov zadosten vnos pa se zagotavlja s prehranskimi dopolnili (PD), v katerih se tradicionalno uporablja sintetična folna kislina. V zadnjih letih je opazen porast uporabe njene aktivne oblike – 5-metiltetrahidrofolat (5- MTHF) v obliki kalcijeve ali glukozaminske soli, ki izkazuje številne prednosti. Ker so PD na trgu manj strogo regulirana v primerjavi z zdravili, smo se v okviru magistrske naloge osredotočili na ugotavljanje vsebnosti in kakovosti PD s 5-MTHF. V ta namen smo uporabili tri predhodno razvite metode (A, B in C), ki smo jih validirali v skladu s smernico ICH. Nadalje smo optimizirali ekstrakcijski postopek ter izbrali topilo, s katerim smo zagotovili topnost in stabilnost pripravljenih vzorcev (10 % metanol, 2 mM natrijev hidroksid, 1 mM etilendiamintetraocetna kislina, 0,24 g/L vitamin C). Optimiziran ekstrakcijski postopek in validirane analizne metode, izbrane na podlagi kompleksnosti vzorcev, smo nato uporabili za analizo izbranih PD. Skupno smo ovrednotili 18 različnih PD s 5-MTHF, s katerimi smo ponazorili trenutno stanje na našem trgu. Prisotnost 5-MTHF smo potrdili pri 16 izdelkih, pri dveh pa je bila vsebnost pod mejo zaznave. Ugotovljeno vsebnost smo primerjali z deklarirano vrednostjo in ocenili skladnosti s smernicami Evropske komisije, ki za PD določajo dopustni interval 80-150 %. Sedem izdelkov (39 %) je bilo pod spodnjo mejo, medtem ko nobeden ni presegal zgornje meje. Izdelke smo nato razvrstili glede na farmacevtsko obliko in obliko 5-MTHF ter ocenili njihov morebitni vpliv na stabilnost oz. dejansko vsebnost vitamina. Analizirali smo 7 tablet, 5 kapsul, 4 tekočine in 2 vrečki, pri čemer so tablete in kapsule izkazale najboljše ujemanje z deklarirano vrednostjo. Pri tekočih izdelkih je bilo največ odstopanja (75,0 % pod spodnjo mejo), kar je najverjetneje posledica nestabilnosti 5-MTHF v vodnem mediju. Pri kalcijevi soli je bilo osem izdelkov (66,7 %) znotraj intervala sprejemljivosti, pri glukozaminski pa dva (40,0 %). Na podlagi tega je težko trditi, katera oblika je intrinzično stabilnejša, saj smo imeli manj izdelkov z glukozaminsko soljo, hkrati pa sta bila dva od teh izdelkov v tekoči obliki. Ovrednotili smo tudi povezavo med ceno in kakovostjo izdelkov, pri čemer smo ugotovili, da večja cena ne zagotavlja boljše kakovosti. Na osnovi dobljenih rezultatov menimo, da je PD s folati treba posvetiti večjo pozornost, saj so tovrstni izdelki oglaševani predvsem nosečnicam. Gre namreč za ranljivo skupino prebivalstva, pri kateri je zadostni vnos folatov še posebej pomemben. Folates or vitamin B9, are crucial for maintaining health, and adequate intake is ensured through food supplements, which traditionally use synthetic folic acid. In recent years, the use of its active form – 5-methyltetrahydrofolate (5-MTHF), as calcium or glucosamine salt, in food supplements is increasing, both of which exhibit several advantages. Since food supplements are less strictly regulated compared to medicinal products, this master’s thesis focused on determining the content and quality of supplements with 5-MTHF. For this purpose, we used three previously developed methods (A, B and C) and validated them in accordance with the ICH guideline. We optimized the extraction procedure and determined a solvent mixture that ensured the stability of prepared samples (10 % methanol, 2 mM sodium hydroxide, 1 mM ethylenediaminetetraacetic acid, 0.24 g/L vitamin C). Subsequently, the optimized extraction procedure and validated analytical methods, chosen based on the complexity of the samples, were used to analyse the selected supplements. We evaluated a total of 18 different supplements containing 5-MTHF, with which we aimed to illustrate the current market situation. The presence of 5-MTHF was confirmed in 16 products, whereas in two products the content was below the detection limit. The determined contents were compared with the declared values and assessed for compliance with the European Commission guidelines, which define an acceptable range of 80–150 %. Seven products (39 %) were below the lower limit, while none exceeded the upper limit. Hereinafter, we classified the products according to their dosage form and type of 5-MTHF salt to evaluate their potential impact on actual content. A total of 7 tablets, 5 capsules, 4 liquids, and 2 sachets were analysed. Tablets and capsules showed the highest conformity with the declared values, while liquid formulations exhibited the highest deviations (75.0 % below 80% of the declared value), likely due to vitamin instability in aqueous media. Among both salts, 8 products containing the calcium form (66.7 %) were within the acceptable range, compared to 2 products (40.0 %) with the glucosamine form. However, it is difficult to conclude which form is intrinsically more stable, due to the lower number of glucosaminebased products, two of which were liquid formulations. Price–quality comparison revealed no direct correlation between higher cost and better quality. Based on the obtained results, it can be concluded that folate-containing food supplements deserve greater attention, as they are primarily marketed to pregnant women – a vulnerable population group for whom adequate folate intake is particularly important. 5-MTHF folna kislina UHPLC vsebnost prehranska dopolnila 5-MTHF folic acid UHPLC content food supplements true false false Slovenski jezik Angleški jezik Magistrsko delo/naloga 2025-12-20 08:45:22 2025-12-20 08:45:36 2025-12-30 03:49:36 0000-00-00 00:00:00 2025 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 1970-01-01 124594 31747.pdf 31747.pdf 1 7AC643956DB93640EE25A7C9EFC5C07C b7e7603c9ceccd7c644c99916acb40c91affe4399a65cbf5e8c9432a3fad0152 7223402f-dd77-11f0-9328-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=224280 Fakulteta za farmacijo 0 0 0