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Razvoj in validacija metode določanja elementov v sledovih v nohtih in laseh
ID Žežlina, Maja (Author), ID Marc, Janja (Mentor) More about this mentor... This link opens in a new window, ID France Štiglic, Alenka (Comentor)

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Abstract
Lasje in nohti so lahko pomembni biološki vzorci za oceno pretekle izpostavljenosti elementom v sledovih, zato se vse pogosteje uporabljajo v kliničnih in epidemioloških raziskavah, v katerih ocenjujejo vpliv okoljskih dejavnikov na zdravje ljudi. Koncentracije elementov v sledovih v Laboratoriju za analitiko encimov in elementov v sledovih, Univerzitetnega kliničnega centra Ljubljana (UKCL), določamo v vzorcih krvi in urina. V okviru magistrske naloge smo razvili in validirali metodo za kvantitativno določanje koncentracij 26 elementov v sledovih v vzorcih las in nohtov, vključno s 7 esencialnimi in 19 neesencialnimi elementi. Na podlagi pregleda literature smo razvili metodo za določanje elementov v sledovih, ki vključuje predanalitske postopke (spiranje, sušenje in kislinski razklop v mikrovalovni pečici) ter analizo tekočih vzorcev z uporabo induktivno sklopljene plazme z masno spektrometrijo (ICP-MS). Metodo smo validirali po smernicah Inštituta za klinične in laboratorijske standarde (CLSI), pri čemer smo določili zmogljivost zaznavanja, natančnost, pravilnost, linearnost in delovno območje metode. Na podlagi validacijskih parametrov smo ugotovili, da metoda izpolnjuje zahteve za predvideno uporabo pri določanju elementov v sledovih v nohtih in laseh. Na vzorcih 140 zdravih oseb obeh spolov smo določili 95 % referenčne intervale za esencialne in neesencialne elemente, ki so bili primerljivi z literaturo, čeprav nekoliko ožji. V nadaljevanju smo ocenili vpliv eksogenih in endogenih dejavnikov ter življenjskih navad in razvad na koncentracije elementov v nohtih na nogi in laseh. Določili smo razlike v koncentracijah V, Mn, Co, Ni, Cu, Zn, As, Se, Sr, Mo, Ag, Cd, Sn med spoloma. S starostjo so se koncentracije večine elementov v vzorcih las in nohtov rahlo nižale. Koncentracije Hg v Obalno-kraški in Goriški regiji so bile povišane, pri ostalih elementih nismo določili pomembnih odstopanj med regijami. Uživanje prehranskih dodatkov je povečalo vsebnost Se v nohtih, uživanje morske hrane pa koncentracije Hg v nohtih in laseh. Kadilci so imeli višje koncentracije Cd in Pb v laseh in nohtih, koncentracije omenjenih elementov pa so korelirale tudi s številom pokajenih cigaret na dan. Lakiranje nohtov ni pokazalo statistično značilnih razlik v koncentraciji elementov, medtem ko je barvanje las vplivalo na koncentracije nekaterih elementov. Mn, Co, Ni, Sr in Ba so bili v umetno barvanih laseh višji, Cu, Zn, As, Se in Mo pa nižji glede na nebarvane. Nazadnje smo primerjali koncentracije elementov v sledovih v nohtih in laseh. Šibko pozitivne korelacije smo določili za elemente Mn, Co, Cu, Se, Al, Ga, As, Sn, Cs, Au V in Pb, močno pozitivno korelacijo pa za element Hg. Nova metoda določanja elementov v sledovih v nohtih in laseh je zanesljiva, vendar so potrebne dodatne usmeritve v standardizacijo in primerljivost rezultatov med različnimi laboratoriji ter opredelitev vpliva barvanja las ter endogenih dejavnikov na večjem številu vzorcev, saj so dosedanji rezultati med študijami nekonsistentni.

Language:Slovenian
Keywords:elementi v sledovih, lasje in nohti, ICP-MS, validacija metode, referenčni intervali
Work type:Master's thesis/paper
Organization:FFA - Faculty of Pharmacy
Year:2024
PID:20.500.12556/RUL-162024 This link opens in a new window
Publication date in RUL:18.09.2024
Views:76
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Secondary language

Language:English
Title:Development and validation of a new method for determining trace elements in nail and hair samples
Abstract:
Hair and nails can serve as important biological samples for assessing past exposure to trace elements, making them increasingly valuable in clinical and epidemiological studies evaluating the effects of environmental factors on human health. In the Laboratory for Enzyme and Trace Element Analytics at the University Medical Centre Ljubljana (UMCL), concentrations of trace elements are typically determined in blood and urine samples. In this master's thesis, we developed and validated a method for the quantitative determination of concentrations of 26 trace elements in hair and nail samples, including 7 essential and 19 non-essential elements. Based on a literature review, we developed a method for trace element determination that incorporates pre-analytical procedures (washing, drying, and acid digestion using a microwave) and liquid sample analysis using inductively coupled plasma mass spectrometry (ICP-MS). The method was validated according to the guidelines of the Clinical and Laboratory Standards Institute (CLSI), assessing detection capability, precision, accuracy, linearity, and working range. Based on the validation parameters, we determined that the method meets the requirements for its intended use in determining trace elements in nails and hair. In samples from 140 healthy individuals of both sexes, we established 95% reference intervals for both essential and non-essential elements, which were comparable to those in the literature, albeit somewhat narrower. We further evaluated the impact of exogenous and endogenous factors on trace element concentrations in toenails and hair. We observed differences in the concentrations of V, Mn, Co, Ni, Cu, Zn, As, Se, Sr, Mo, Ag, Cd, and Sn between genders. With age, the concentrations of most elements in hair and nail samples slightly decreased. Hg concentrations were elevated in the Coastal-Karst and Goriška regions, while no significant regional differences were observed for other elements. Supplement intake increased Se content in nails, and seafood consumption raised Hg concentrations in both nails and hair. Smokers had higher concentrations of Cd and Pb in their hair and nails, with these elements correlating with the number of cigarettes smoked per day. Nail polish did not show statistically significant differences in element concentrations, whereas hair dyeing affected the concentrations of certain elements. Mn, Co, Ni, Sr, and Ba were higher in dyed hair, while Cu, Zn, As, Se, and Mo were lower compared to undyed hair. Finally, we compared trace element concentrations in nails and hair. Weak positive correlations were found for Mn, Co, Cu, Se, Al, Ga, As, Sn, Cs, Au, V, and Pb, while a strong positive correlation was observed for Hg. The newly developed method for determining trace elements in nails and hair is reliable, although further efforts are needed to standardize and harmonize results between different laboratories, as well as to better understand the influence of hair dyeing and endogenous factors based on a larger number of samples, as current study results have been inconsistent.

Keywords:trace elements, hair and toenails, ICP-MS, method validation, reference intervals

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