Desenvolvimento de metodologia para avaliação de ácidos graxos em amostras de sêmen humano para fins de estudo de fertilidade
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2022-02-22
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Universidade Federal de Goiás
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Infertility has been the subject of studies by several researchers around the world, in this field, the influence of diet has been one of the factors evaluated. In women, the cell responsible for reproduction is the egg, for men, it is the sperm, present in human semen, which is a complex matrix, which contains proteins, amino acids, lipids, among other compounds. A semen, with desirable qualities, needs to demonstrate good concentration, morphology, vitality and motility of the sperm present. The lipids present in the composition, especially the fatty acids, such as omegas 3, 6 and 9, help in the structuring of cell membranes, making them more resistant, also acting in the improvement of motility. Thus, in partnership with the Department of Biomedicine of the Federal University of Jataí (UFJ), under number 3,792,715 of the ethics committee of the Federal University of Goiás (UFG), this work proposes to evaluate possible relationships between seminal quality and ingestion of baru nut, typical food of the cerrado region of Goiás. For analytical purposes, the Mass Spectrometry (MS) technique was used, this is a technique subject to the presence of endogenous compounds of the complex matrix, such as semen. Therefore, the sample preparation step for analysis is extremely important. The miniaturized sample preparation technique, disposable pippete extraction (DPX), was evaluated in terms of cleaning the sample and extracting the analytes of interest. In order to increase the selectivity of the technique, a DPX extractor phase, developed by our research group, was applied to extract fatty acids from human semen, followed by analysis by direct infusion in the mass spectrometer. The parameters with direct influence on the efficiency of the DPX technique were optimized. The results showed greater extraction efficiency for 700 μL of semen sample aspirated/dispensed for 7 cycles, followed by desorption, using 1000 μL of acetonitrile in just 1 cycle. For comparison purposes, the samples were also analyzed with sample preparation performed by the adapted Bligh & Dyer method, followed by direct infusion in high resolution mass spectrometry. The method developed was submitted to the evaluation of selectivity, matrix effect, calibration curve, precision and accuracy, following the current legislation for bioassays, RDC 27/2012, drafted and supervised by the National Health Surveillance Agency (ANVISA).
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BUENO, A. C. Desenvolvimento de metodologia para avaliação de ácidos graxos em amostras de sêmen humano para fins de estudo de fertilidade. 2022. 72 f. Dissertação (Mestrado em Química) - Universidade Federal de Goiás, Goiânia, 2022.