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Here we developed a powerful tool for comprehensive data collection and mapping of molecular and elemental signatures in the Melanoma-bearing Libechov Minipig (MeLiM) model. The combination of different mass spectrometric methods allowed for detail investigation of specific melanoma markers and elements and their spatial distribution in tissue sections. MALDI-MSI combined with HPLC-MS/MS analyses resulted in identification of seven specific proteins, S100A12, CD163, MMP-2, galectin-1, tenascin, resistin and PCNA that were presented in the melanoma signatures. Furthermore, the ICP-MS method allowed for spatial detection of zinc, calcium, copper, and iron elements linked with the allocation of the specific binding proteins.
In this study, carbon microparticles (CMs) as a physical carrier to enhance the analyte transport efficiency for inductively coupled plasma mass spectrometry connected with electrothermal vaporization (ETV-ICP-MS) are proposed. Carbon microparticles mixed with samples or calibration standard solutions were dosed as a slurry into the graphite furnace. The optimization of working conditions was done for the standard solution 0.5 μg L-1 of Au and Tl. The pyrolysis and vaporization temperatures were 500 and 2700 °C for Au and 400 and 1900 °C for Tl. The optimized CMs concentration was 1 g L-1 for Au and 2.5 g L-1 for Tl. For the quantification, external calibration standard solutions were used. The result obtained for Au in the digested CRM GBW 07601 (the informatory value 2.1 ± 2 μg kg-1) was 2.2 ± 0.1 μg kg-1. The result for Tl in the digested CRM BCR 679 (the certified value 3 ± 0.3 μg kg-1) was 3.2 ± 0.2 μg kg-1 and in the slurry 2.7 ± 0.1 μg kg-1. The result for Tl in the CRM GBW 10052 (the certified value 57 ± 11 μg kg-1) was 51 ± 3 μg kg-1. The instrumental limits of detection were 0.016 ng L-1 for Au and 0.026 ng L-1 for Tl. The recoveries and repeatabilities measured on calibration standards were in the range 99-100% and 0.2-2.3% for Au and 100-111% and 2.9-6.7% for Tl.
- Publikační typ
- časopisecké články MeSH
- MeSH
- hliník aplikace a dávkování metabolismus škodlivé účinky MeSH
- kovy chemie MeSH
- lidé MeSH
- měď aplikace a dávkování metabolismus MeSH
- odběr biologického vzorku MeSH
- spektrofotometrie atomová metody přístrojové vybavení MeSH
- stopové prvky analýza chemie MeSH
- zinek aplikace a dávkování metabolismus škodlivé účinky MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- srovnávací studie MeSH
The presented work proposes a novel analytical ICP-MS-based approach for the accurate and precise chromium speciation in biological tissues. The determination of total Cr(VI) and soluble Cr(III) species was carried out by alkaline EDTA extraction followed by their separation using ion-exchange high-performance liquid chromatography inductively coupled plasma mass spectrometry (IE-HPLC-ICP-MS). The developed method was validated according to the procedure given in the United States Food and Drug Administration guideline on the validation of bioanalytical methods. Validation parameters included limit of detection (≤ 0.03 μg g-1), limit of quantification (≤ 0.08 μg g-1), linearity (r ≥ 0.9998), intra-day and inter-day accuracy (86-110%) and precision (≤ 10%), extraction recovery (89-110%), carry-over effect and sensitivity. In addition, special attention was paid to the study of chromium species interconversion and the elimination of spectral interferences. Moreover, the validated ICP-MS method employing microwave acid digestion was used to determine the total Cr content in collected fractions. Finally, the whole ICP-MS-based methodology was applied to the analyses of two certified reference materials of hepatopancreas tissue. Obtained results indicated that the majority of chromium in biological tissues is bound to the solid residue, Cr(VI) was determined in none of the samples investigated. This is the first study focusing on soluble Cr(III), total Cr(VI), and total bound Cr species in biological tissues. It is characterized by efficient sample preparation and fast simultaneous analysis of Cr species with parallel total Cr analysis serving for chromium balance evaluation.
- MeSH
- chemie analytická * trendy MeSH
- chrom analýza izolace a purifikace MeSH
- kadmium analýza izolace a purifikace MeSH
- krev MeSH
- krevní plazma chemie MeSH
- lidé MeSH
- moč MeSH
- nikl analýza izolace a purifikace MeSH
- olovo analýza izolace a purifikace MeSH
- spektrofotometrie atomová * metody přístrojové vybavení využití MeSH
- statistika jako téma MeSH
- tělesné tekutiny * chemie metabolismus MeSH
- těžké kovy analýza izolace a purifikace MeSH
- Check Tag
- lidé MeSH
