Vrednotenje vsebnosti nitrozaminov v zdravilih, prehranskih dopolnilih in kozmetičnih izdelkih

Planinšek Parfant, Timeja (author), Roškar, Robert (mentor)

N-nitrozamini kot genotoksične nečistote predstavljajo v zadnjih letih eno izmed osrednjih problematik farmacevtske industrije. Zato smo v okviru doktorske naloge celovito obravnavali problematiko N-nitrozaminov v zdravilih, prehranskih dopolnilih in kozmetičnih izdelkih ter izvedli oceno izpostavljenosti N-nitrozaminom. Najprej smo razvili celovito analizno metodologijo za sočasno določanje številnih N-nitrozaminov, ki temelji na tekočinski kromatografiji sklopljeni z masno spektrometrijo visoke ločljivosti ter napredni pripravi vzorca. Razvili smo štiri inovativne in sodobne analizne pristope, ki izkazujejo visoko občutljivost, selektivnost, robustnost in točnost ter omogočajo določanje doslej največjega števila reguliranih nizkomolekularnih N-nitrozaminov v zdravilih in kozmetičnih izdelkih ter novih N-nitrozaminov, ki so strukturno sorodni vitaminom, v zdravilih in prehranskih dopolnilih. Analizne metode smo implementirali na 215 različnih izdelkov (51 zdravil, 53 kozmetičnih izdelkov in 111 prehranskih dopolnil) na slovenskem trgu. Ugotovili smo, da so N-nitrozamini prisotni v teh izdelkih, v zdravilih le v omejenem obsegu in skladno z regulativnimi zahtevami, medtem ko smo N-nitrozamine zaznali v 30 % kozmetičnih izdelkov, pri čemer je 15 % izdelkov presegalo regulativno mejo do približno trikrat. N-nitrozamine, ki so strukturno sorodni vitaminom, smo zaznali v 43 % prehranskih dopolnil, pri čemer so trije izdelki presegali regulativno mejo, veljavno za zdravila. Posebno pozornost smo namenili tudi proučevanju razlogov za pojavnost N-nitrozaminov v kozmetičnih izdelkih in prehranskih dopolnilih ter identificirali strukturne značilnosti in vsebnost predhodnikov N-nitrozaminov ter nitritov kot ključne dejavnike njihovega nastanka med shranjevanjem izdelkov. Kot prvi smo v teh izdelkih zaznali nov N-etil-N-(2-hidroksietil) nitrozamin. V zaključnem delu doktorske naloge smo s sistematičnim pregledom literature in uporabo lastnih podatkov o njihovi vsebnosti kot prvi izvedli celovito oceno dnevne izpostavljenosti preko različnih virov z upoštevanjem njihovega genotoksičnega potenciala. Pokazali smo, da predstavljajo tobačni izdelki največji vir vnosa (68 %), sledijo živila (16 %) in kozmetični izdelki (12 %), v zanemarljivem deležu pa prispevajo zdravila, prehranska dopolnila in pitna voda.

• Language: Slovenian
• Keywords: N-nitrozamini, genotoksične nečistote, analizne metode, masna spektrometrija visoke ločljivosti, ocena izpostavljenosti, nastanek N-nitrozaminov, stabilnostna študija
• Work type: Doctoral dissertation
• Typology: 2.08 - Doctoral Dissertation
• Organization: FFA - Faculty of Pharmacy
• Publisher: [T. Planinšek Parfant]
• Year: 2026
• PID: 20.500.12556/RUL-188410
• UDC: 543.544.5:543.51(043.3)
• COBISS.SI-ID: 292691715
• Publication date in RUL: 23.09.2026

Secondary language

• Language: English
• Title: Evaluation of nitrosamine content in pharmaceuticals, food supplements and cosmetic products

Abstract

N-nitrosamines are genotoxic impurities that have become the focus of the pharmaceutical industry in recent years. This doctoral thesis comprehensively addresses N-nitrosamine-related challenges in medicines, food supplements and cosmetics. The comprehensive analytical methodology for the simultaneous determination of multiple N-nitrosamines was developed, combining advanced sample preparation with liquid chromatography coupled to high resolution mass spectrometry. Four innovative state-of-the-art analytical approaches with high sensitivity, selectivity, robustness, and accuracy were established for the determination of the largest number of regulated low-molecular-weight N-nitrosamines reported to date in medicines and cosmetics, as well as emerging N-nitrosamine drug substance-related impurities (NDSRIs) in medicines and food supplements. Analytical methods were further applied to 215 different products (51 medicines, 53 cosmetics, 111 food supplements) on the Slovenian market. Our findings show that N-nitrosamines are present in these products, with limited occurrence in medicines complying with current regulatory requirements, whereas N-nitrosamines were detected in 30% of cosmetics, with 15% exceeding the regulatory limit by up to threefold. NDSRIs were detected in 43% of food supplements, with three products exceeding the regulatory limit applicable to medicines. Special attention was paid to investigating the factors contributing to N-nitrosamine occurrence in cosmetics and food supplements. Structural characteristics, the content of precursors and nitrites were identified as key factors in N nitrosamine formation during storage. Furthermore, the emerging N-ethyl-N-(2-hydroxyethyl) nitrosamine was identified in these products for the first time. In the final part of the thesis, a comprehensive assessment of daily N-nitrosamine exposure from multiple sources was performed for the first time, combining a systematic literature review with our own data on N-nitrosamine occurrence and considering their genotoxic potential. It was demonstrated that tobacco products are the predominant source of exposure (68%), followed by food (16%) and cosmetics (12%), whereas medicines, food supplements and drinking water made only a negligible contribution to total exposure.

• Keywords: N-nitrosamines, genotoxic impurities, analytical methods, high-resolution mass spectrometry, exposure assessment, N-nitrosamine formation, stability study