A method development for measuring lithium uptake inCaco-2 cells in a complex matrix using MP-AES – Appliedto evaluate the impact of humic acids on lithium uptake

University essay from Örebro universitet/Institutionen för naturvetenskap och teknik

Author: Erica Hjelm; [2019]

Keywords: Lithium; humic substances; MP AES; Caco-2;

Abstract: Lithium, together with hydrogen and helium, were the only elements formed duringthe big bang. Lithium today, is a component in various products, some examples aregrease, ceramics, lithium-ion batteries and medication for bipolar disorder. Eventhough the knowledge of lithium’s interactions with biota and our environment islimited, the use of lithium is increasing. In humans, lithium is primarily absorbed in thesmall intestine through Na-channels. The average daily intake varies a lot, but in1985 it was estimated to range from 650 to 3100 μg for a 70 kg adult, according tointernational studies. However, it is not considered as a micronutrient and does nothave a recommended daily intake. Levels in some drinking waters are estimated toreach 170 μg L-1but no definite limit values are set. Humic substances are omnipresent in soil and freshwaters and is well known to formcomplexes with various cations such as most metals. In plants, the complexation ofcadmium and zinc with humic substances decreases their toxicity. However,currently, no studies have been published regarding if this is valid for animal cells aswell. To investigate if this is the case, in vitro tests were performed on Caco-2 cells,which originate from a colon carcinoma. These cells were exposed to a lithium concentration of 10 μg L-1in a nutrient medium for 2 hours. To a series of thesamples, humic acid was added to give a final concentration of 9.86 μg L-1 in thesolution to investigate if that would influence the lithium uptake. The study also includes a methodological evaluation if the performance of microplasma atomic emission spectroscopy (MP-AES) is suitable for the analyses of heavymatrices, e.g. nutrient medium and 1% Triton X solution. After optimization of instrumental parameters, it was concluded that MP-AES can beused for analysis of heavy and complex matrices. It requires, however, the use ofmatrix matched calibration solutions and addition of caesium as ionization buffer incombination with lithium signal correction using internal standards. It is alsoconcluded that lithium is absorbed by Caco-2 cells and that there was no indicationthat humic acid altered this uptake.

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