Volume: 3, 2024
3rd International PhD Student’s Conference at the University of Life Sciences in Lublin, Poland:
ENVIRONMENT – PLANT – ANIMAL – PRODUCT
Abstract number: H016
DOI: https://doi.org/10.24326/ICDSUPL3.H016
Published online: 24 April 2024
ICDSUPL, 3, H016 (2024)
Optimization of cheese preparation procedures for vitamin determination using the ELISA method
Anna Lipian-Głos1*, Anna Stępniowska1, Paulina Osowska1, Agnieszka Serek2, Magdalena Krauze1, Katarzyna Ognik1, Bartosz Sołowiej3
1 Department of Biochemistry and Toxicology, Faculty of Animal Sciences and Bioeconomy, University of Life Sciences in Lublin, Akademicka 13, 20-950 Lublin, Poland
2 Biochemstry Research Club, University of Life Sciences in Lublin, Akademicka 13, 20-950 Lublin, Poland
3 Department of Dairy Technology and Functional Foods, Faculty of Food Sciences and Biotechnology, University of Life Sciences in Lublin, Skromna 8, 20-704 Lublin, Poland
* Corresponding author: anna.lipian@up.lublin.pl
Abstract
Cheese is a rich source of fat, which may cause interference during vitamin determinations. Therefore, an important stage of sample preparation is the separation of the protein part from the fat part, which can be achieved by using liquid extraction (LLE). The aim of the work was to optimize the preparation of cheese samples using LLE for the determination of selected B vitamins (B2, B6, B12) and fat-soluble vitamins (A, D, E). In the first stage of sample preparation, 1.5 or 3 mL of buffered saline with pH = 7.4 (PBS) was added to a cheese weighing (approx. 0.1 g), and then the samples were homogenized using a mechanical homogenizer. Then 2, 3 or 4 mL of chloroform were added to the homogenates and placed under ultrasound for 5 min. The samples were then centrifuged at 4500 g for 5 min. After centrifugation, two separate phases were obtained: aqueous and chloroform. A chloroform layer was taken for the determination of vitamins A, D and E, and an aqueous layer was taken for the determination of B vitamins. The samples prepared in this way were placed on an ELISA plate for the determination of the tested vitamins. The analysis was performed according to the manufacturer’s procedure. Analysis of the obtained results allowed for the conclusion that the best results were obtained using 3 mL of PBS and 3 mL of chloroform, and such volumes of these reagents were selected for further tests.
Task titled: “Research network of natural science universities for the development of the Polish dairy sector – research project” is financed under a targeted subsidy from the Minister of Education and Science (No. MEiN/2023/DPI/2862).
Keywords: cheese, B vitamins, vitamin A, vitamin D
How to cite
A. Lipian-Głos, A. Stępniowska, P. Osowska, A. Serek, M. Krauze, K. Ognik, B. Sołowiej, 2024. Optimization of cheese preparation procedures for vitamin determination using the ELISA methods. In: 3rd International PhD Student’s Conference at the University of Life Sciences in Lublin, Poland: Environment – Plant – Animal – Product. https://doi.org/10.24326/ICDSUPL3.H016