Določanje referenčnih intervalov elementov v sledovih v krvi zdravih odraslih oseb z metodo masne spektrometrije z induktivno sklopljeno plazmo

Žavbi, Marko (author), Skitek, Milan (mentor), France - Štiglic, Alenka (co-mentor)

Na Kliničnem inštitutu za klinično kemijo in biokemijo smo v Laboratoriju za analitiko encimov in elementov v sledovih uvedli sodobnejšo tehnologijo za analizo elementov v sledovih - masno spektrometrijo z induktivno sklopljeno plazmo (ICP-MS). ICP-MS je najsodobnejša tehnologija za več-elementarno analizo bioloških materialov, ki jo karakterizirajo izjemna občutljivost, specifičnost in ponovljivost. Nova tehnologija je laboratoriju omogočila razširitev nabora analiziranih elementov v sledovih in zahtevala posodobitev obstoječih referenčnih intervalov ter postavitve novih referenčnih intervalov. Za določitev zanesljivih referenčnih intervalov smo z izborom predanalitičnih in analitičnih postopkov zagotovili primerno odraslo slovensko populacijo. Pri izvajanju protokola analize smo sledili standardnim smernicam CLSI-C28-A3. V vzorcih polne krvi smo z metodo redčenja v amonijevi raztopini in tehnologijo ICP-MS določali 27 elementov v sledovih: Li, Rb, Cs, Be, Sr, Ba, Ga, Tl, Sn, Pb, As, Sb, Se, Ti, V, Cr, Mo, Mn, Co, Ni, Cu, Ag, Au, Zn, Cd, Hg in U. Določili smo referenčne intervale za elemente v sledovih v krvi za odraslo slovensko populacijo: Litij (Li) < 6.5 µg/l, Rubidij (Rb) 1597-3529 µg/l, Cezij (Cs) < 6.0 µg/l, Berilij (Be) < 0.271 µg/l, Stroncij (Sr) < 21 µg/l, Barij (Ba) < 0.74 µg/l, Galij (Ga) < 0.059 µg/l, Talij (Tl) < 0.034 µg/l, Kositer (Sn) < 0.62 µg/l, Svinec (Pb) < 29 µg/l, Arzen (As) < 1.64 µg/l, Antimon (Sb) ni določen, Selen (Se) 71-156 µg/l, Titan (Ti) ni določen, Vanadij (V) < 0.070 µg/l, Krom (Cr) < 0.45 µg/l, Molibden (Mo) < 1.0 µg/l, Mangan (Mn) 4.2-14.1 µg/l, Kobalt (Co) < 0.33 µg/l, Nikelj (Ni) < 0.25 µg/l, Baker (Cu) 508-1307 µg/l, Srebro (Ag) < 0.232 µg/l, Zlato (Au) < 0.008 µg/l, Cink (Zn) 5944-7600 µg/l, Kadmij (Cd) < 1.49 µg/l, Živo srebro (Hg) < 4.6 µg/l in Uran (U) < 0.019 µg/l. Za zanesljivo določanje Sb in Ti smo razširili standardni rutinski postopek določanja. Rezultate referenčnih intervalov slovenske populacije smo primerjali z dostopnimi svetovnimi viri podatkov in ovrednotili ujemanja in morebitna odstopanja. S primerjavo smo ugotovili, da na razlike med referenčnimi intervali za elemente v sledovih med primerjanimi populacijami pomembno vplivajo geografski, ekonomski in individualni dejavniki. Ugotavljamo tudi, da je za interpretacijo slovenske populacije nezanesljivo povzemati in uporabljati referenčne intervale tujih populacij. Z našo raziskavo smo potrdili izredno prednost in pomembnost določitve lastnih referenčnih intervalov za elemente v sledovih, saj so le zanesljivi referenčni intervali edini pravilni pristop v interpretaciji laboratorijskih rezultatov.

• Language: Slovenian
• Keywords: Elementi v sledovih, referenčni intervali, CLSI-C28-A3, ICP-MS, induktivno sklopljena plazma, masna spektrometrija, človek, kri
• Work type: Master's thesis/paper
• Organization: FFA - Faculty of Pharmacy
• Year: 2020
• PID: 20.500.12556/RUL-120595
• Publication date in RUL: 23.09.2020

Secondary language

• Language: English
• Title: Determination of trace elements' reference intervals in healthy adults' blood samples by inductively coupled plasma mass spectometry

Abstract

At the Clinical Institute of Clinical Chemistry and Biochemistry, in the Laboratory for Analysis of Enzymes and Trace Elements we have introduced a modern technology for analysis of trace elements - Inductively Coupled Plasma Mass Spectrometry (ICP-MS). ICP-MS is a state-of-the-art technology for multi-elemental analysis of biological materials, characterised by great sensitivity, specificity and repeatability. New technology has enabled the laboratory to expand the set of analysed trace elements and required the updating of existing reference intervals and the setting of new reference intervals. To determine reliable reference intervals, we ensured a suitable adult Slovenian population by selecting pre-analytical and analytical procedures. The standard CLSI-C28-A3 guidelines were followed when performing the analysis protocol. In whole blood samples, 27 trace elements were determined by the dilution method in ammonium solution and ICP-MS technology: Li, Rb, Cs, Be, Sr, Ba, Ga, Tl, Sn, Pb, As, Sb, Se, Ti, V, Cr, Mo, Mn, Co, Ni, Cu, Ag, Au, Zn, Cd, Hg and U. We determined reference intervals for trace elements in the blood of the adult Slovenian population: Lithium (Li) < 6.5 µg/l, Rubidium (Rb) 1597- 3529 µg/l, Caesium (Cs) < 6.0 µg/l, Beryllium (Be) < 0.271 µg/l, Strontium (Sr) < 21 µg/l, Barium (Ba) < 0.74 µg/l, Gallium (Ga) < 0.059 µg/l, Thallium (Tl) < 0.034 µg/l, Tin (Sn) < 0.62 µg/l, Lead (Pb) < 29 µg/l, Arsenic (As) < 1.64 µg/l, Antimony (Sb) not determined, Selenium (Se) 71-156 µg/l, Titanium (Ti) not determined, Vanadium (V) < 0.070 µg/l, Chromium (Cr) < 0.45 µg/l, Molybdenum (Mo) < 1.0 µg/l, Manganese (Mn) 4.2-14.1 µg/l, Cobalt (Co) < 0.33 µg/l, Nickel (Ni) < 0.25 µg/l, Copper (Cu) 508-1307 µg/l, Silver (Ag) < 0.232 µg/l, Gold (Au) < 0.008 µg/l, Zinc (Zn) 5944-7600 µg/l, Cadmium (Cd) < 1.49 µg/l, Mercury (Hg) < 4.6 µg/l and Uranium (U) < 0.019 µg/l. For reliable determination of Sb and Ti, we extended the standard routine protocol. The results of the reference intervals of the Slovenian population were compared with available global data sources and evaluated for matches and possible deviations. By comparison, we found that the differences between the reference intervals for trace elements between the compared populations were significantly influenced by geographical, economic and individual factors. We found that for the interpretation of the Slovenian population it is unreliable to summarise and use reference intervals of foreign populations With our research, we confirmed the extraordinary advantage and importance of determining our own reference intervals for trace elements, as only reliable reference intervals are the only correct approach in the interpretation of laboratory results.

• Keywords: Trace elements, reference intervals, CLSI-C28-A3, ICP-MS, inductively coupled plasma, mass spectrometry, human, blood