Volume: 3, 2024
3rd International PhD Student’s Conference at the University of Life Sciences in Lublin, Poland:
ENVIRONMENT – PLANT – ANIMAL – PRODUCT
Abstract number: H017
DOI: https://doi.org/10.24326/ICDSUPL3.H017
Published online: 24 April 2024
ICDSUPL, 3, H017 (2024)
Optimization of procedures for preparing buttermilk and protein milk shake samples for vitamin determination using the ELISA method
Paulina Osowska1*, Anna Stępniowska1, Jagoda Szafrańska2, Magdalena Krauze1, Bartosz Sołowiej2, Katarzyna Ognik1
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 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: paulina.osowska@up.lublin.pl
Abstract
Buttermilk and protein milk shake drinks are dairy products with a high protein content and therefore a complex matrix. For this reason, the procedure for preparing this type of samples should be optimized accordingly. The aim of the work was to optimize the preparation of buttermilk and protein milk shake samples for the determination of selected B vitamins (B2, B6, B12) and fat-soluble vitamins (A, D, E). The tested products were placed in an ultrasonic bath for 5 minutes to homogenize the samples. Then, product weights (0.1 or 0.25 g) were homogenized with 3 mL of buffered saline with pH = 7.4 (PBS) using a mechanical homogenizer. In the next stage, it was checked whether the use of liquid-liquid extraction would reduce interference from proteins. 1.5 or 3 mL of chloroform (a non-polar solvent, immiscible with water) was added to the homogenates and the samples were placed in ultrasound for 5 min. The samples were then centrifuged at 4500 g for 5 min and two phases were obtained. A chloroform layer was taken for the determination of fat-soluble vitamins, and an aqueous layer was taken for the determination of water-soluble vitamins. The samples prepared in this way were placed on an ELISA plate to determine individual vitamins. The analysis was performed according to the manufacturer’s procedure. The analysis of the obtained results allowed the conclusion that for the buttermilk and protein milk shake samples, liquid-liquid extraction was not necessary, therefore, for the determination of vitamins using the ELISA method, the samples were prepared by homogenizing the product with PBS.
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: buttermilk, B vitamins, vitamin A, vitamin D
How to cite
P. Osowska, A. Stępniowska, J. Szafrańska, M. Krauze, B. Sołowiej, K. Ognik, 2024. Optimization of procedures for preparing buttermilk and protein milk shake samples for vitamin determination using the ELISA method. 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.H017