ORIGINAL PAPER
Improved lipid saponification for chromatographic quantification
of fatty acids in porcine erythrocytes – an important lipidomic
biomarker of the effectiveness of dietary fat supplementation
in pigs as a large animal model for human studies
K. Pierzynowska
1,2,3
1
The Kielanowski Institute of Animal Physiology and Nutrition, Polish Academy of Sciences, 05-110 Jabłonna, Poland
2
Witold Chodźko Institute of Rural Health, Department of Medical Biology, 20-090 Lublin, Poland
3
Lund University, Department of Biology, Sölvegatan 35, 22362 Lund, Sweden
Publication date: 2023-06-13
Corresponding author
M. Czauderna
The Kielanowski Institute of Animal Physiology and Nutrition, Polish Academy of Sciences, 05-110 Jabłonna, Poland
The Kielanowski Institute of Animal Physiology and Nutrition, Polish Academy of Sciences, 05-110 Jabłonna, Poland
J. Anim. Feed Sci. 2023;32(4):385-399
KEYWORDS
TOPICS
ABSTRACT
The objectives of our study were to enhance and evaluate an
improved saponification method for the quantification of fatty acids (FA), with
a specific focus on highly-unsaturated long-chain polyunsaturated FA (LPUFA)
that are easily peroxidised in pig erythrocytes. These erythrocytes serve as
a valuable large animal model for human studies. We implemented a modified
saponification procedure, involving a shorter initial saponification at 95 °C
for 2 min, followed by overnight saponification at 22–25 °C, and subsequent
gentle base and acid-catalysed methylation of FA. These analytical procedures
proved to be suitable for gas-chromatographic analysis of highly-unsaturated
LPUFA, which are prone to peroxidation in processed piglet erythrocytes. The
modified initial saponification at 95 °C, followed by overnight saponification at
22–25 °C, and subsequent methylation, were used to assess the suitability
of erythrocytes for the determination of FA, tocopherol and malondialdehyde
(MDA) concentrations in monogastric mammalian tissues. The content of
docosahexaenoic acid (DHA) in erythrocytes can serve as a marker for DHA
bioaccumulation in animal tissues and dietary contents of n-3LPUFA (especially
DHA). The concentrations of δ-, γ-, α-tocopherols, total cholesterol and MDA
increased in piglet erythrocytes during 16 days of exposure to dietary fat.
The levels of FA and tocopherols can therefore be used in erythrocytes as
indicators of the bioaccumulation yield of these compounds in animal tissues.
Additionally, we propose that FA and tocopherol profiles in erythrocytes may be
used as indicators of the concentrations of these components in diets and the
effectiveness of their bioaccumulation in animal tissues.
ACKNOWLEDGEMENTS
The authors would like to express their gratitude to Dr. Janine Donaldson for her valuable contributions and feedback during the preparation of the manuscript. Her insights and expertise greatly enhanced the quality of this research.
FUNDING
We would like to acknowledge the financial support provided by the National Science Centre, Grant No. 2020/04/X/NZ9/00358 (MINIATURA 4), as well as the support from the statutory funds of The Kielanowski Institute of Animal Physiology and Nutrition, Polish Academy of Sciences, Jabłonna, Poland.
CONFLICT OF INTEREST
The Authors declare that there is no conflict of interest.
REFERENCES (36)
1.
Ahmed M., Moazzami A.A., Andersson R., Pickova J., 2011. Varying quality of fish oil capsules: fatty acids and tocopherol. Neuro. Endocrinol. Lett. 32, 37–40, https://pubmed.ncbi.nlm.nih.go....
2.
AOAC., 2005. Current through revision 4. AOAC International, Suite. In: Association of official analytical chemist, official methods of analysis. 18th edition. vol. 500; 2005. p. 481. North Frederick Avenue, Gaithersburg, Maryland 20877–2417 (USA).
3.
Baur L.A., O’Connor J., Pan D.A., Wu B.J., O’Connor M.J., Storlien L.H., 2000. Reltionships between the fatty acid composition of muscle and erythrocyte membrane phospholipids in young children and the effect of type of infant feeding. Lipids 35, 77–82, https://doi.org/10.1016/s0026-....
4.
Baylin A., Campos H., 2006. The use of fatty acid biomarkers to reflect dietary intake. Curr. Opin. Lipidol. 17, 22–27, https://doi.org/10.1097/01.mol....
5.
Boccardo A., Compiani R., Baldi G., Pravettoni D., Grossi S., Sala G., Taylor S., Neville E., Sgoifo Rossi C.A., 2022. Effects of a supplemental calcareous marine algae bolus on blood calcium concentration in dairy heifers. J. Anim. Feed Sci. 31, 40–45, https://doi.org/10.22358/jafs/....
6.
Bystrická Z., Ďuračková Z., 2016. Gas chromatography determination of fatty acids in the human erythrocyte membranes - a review. PLEFA 115, 35–40, https://doi.org/10.1016/j.plef....
7.
Callahan A., Leonard H., Powell T., 2020. Nutrition: Science and everyday application. Powered by Pressbooks; October 14, 2020; E-book, ISBN: 978-1-63635-003-5.
8.
Czauderna M., Kowalczyk J., Niedźwiedzka K.M., Wąsowska I., Pastuszewska B., Bulska E., Ruszczyńska A., 2004. Liver and body mass gain, content of conjugated linoleic acid (CLA) isomers and other fatty acids in the liver of rats fed CLA isomers and selenium. J. Anim. Feed Sci. 13, 353–369, https://doi.org/10.22358/jafs/....
9.
Czauderna M., Kowalczyk J., Korniluk K., Wąsowska I., 2007. Improved saponification then mild base and acid-catalyzed methylation is a useful method for quantifying fatty acids, with special emphasis on conjugated dienes. Acta Chromatogr. 18, 59–71, Corpus ID: 26464712.
10.
Czauderna M., Kowalczyk J., Marounek M., 2011. The simple and sensitive measurement of malondialdehyde in selected specimens of biological origin and some feed by reversed phase high performance liquid chromatography. J. Chromatogr. B 879, 2251–2258. https://doi.org/10.1016/j.jchr....
11.
Czauderna M., Kowalczyk J., Niedźwiedzka K.M., 2009. Simple HPLC analysis of tocopherols and cholesterol from specimens of animal origin. Chem. Anal. (Warsaw) 54, 203–214, https://www.researchgate.net/p....
12.
Detopoulou P., Fragopoulou E., Alepoudea E., Nomikos T., Kalogeropoulos N., Antonopoulou S., 2018. Associations between erythrocyte fatty acids and Mediterranean diet in Greek volunteers. Hell. J. Atheroscler. 9, 17–31, https://doi.org/10.23803/HJA.V....
13.
Di Marino L., Maffettone A., Cipriano P., Sacco M., Di Palma R., Amato B., Quarto G., Riccardi G., Rivellese A.A. 2000. Is the erythrocyte membrane fatty acid composition a valid index of skeletal muscle membrane fatty acid composition? Metab. Clin. Exper. 49(9), 1164–1166, https://doi.org/10.1053/META.2....
14.
Ding D., Li Y.H., Xiao M.L., Dong H.L., Lin J.S., Chen G.D., Chen Z.Y., Tang X.Y., Chen Y.M., 2020. Erythrocyte membrane polyunsaturated fatty acids are associated with incidence of metabolic syndrome in middle-aged and elderly people-an 8.8-year prospective study. J. Nutr. 150(6), 1488–1498, https://doi.org/10.1093/jn/nxa....
15.
Enriquez Y.R., Giri, M., Rottiers, R., Christophe A., 2004. Fatty acid composition of erythrocyte phospholipids is related to insulin levels, secretion and resistance in obese type 2 diabetics on Metformin. Clin. Chim. Acta 346(2), 145–152, https://doi.org/10.1016/j.cccn....
16.
Gabriel M., Azucena M., Alejandra S., Jorge M., 2013. Association between metabolic syndrome and erythrocyte fatty acid profile in Mexican adolescents: a trans fatty acid approach. Food Nutr. Sci. 4(9A), 51–58, https://doi.org/10.4236/fns.20....
17.
Goncharova K., Pierzynowski S.G., Grujic D., Kirko S., Szwiec K., Wang J., Kovalenko T., Osadchenko I., Ushakova G., Shmigel H., Fedkiv O., Majda B., Prykhodko O., 2014. A piglet with surgically induced exocrine pancreatic insufficiency as an animal model of newborns to study fat digestion. Br. J. Nutr. 112(12), 2060–2067, https://doi.org/10.1017/S00071....
18.
Harris H.R., Eliassen A.H., Doody D.R., Terry K.L., Missmer S.A., 2020. Dietary fat intake, erythrocyte fatty acids, and risk of uterine fibroids. Fertil. Steril. 114(4), 837–847, https://doi.org/10.1016/j.fert....
19.
Jo S.H., Han S.H., Kim S.H., Eckel R.H., Koh K.K., 2021. Cardiovascular effects of omega-3 fatty acids: Hope or hype? Atherosclerosis 322, 15–23, https://doi.org/10.1016/j.athe....
20.
Kabagambe E.K., Tsai M.Y., Hopkins P.N., Ordovas J.M., Peacock J.M., Borecki I.B., Arnett D.K., 2008. Erythrocyte fatty acid composition and the metabolic syndrome: a National Heart, Lung, and Blood Institute GOLDN study. Clin. Chem. 54(1), 154–162, https://doi.org/10.1373/clinch....
21.
Kawabata T., Hirota S., Hirayama T., Adachi N., Kaneko Y.,Iwama N., Kamachi K., Araki E., Kawashima H., Kiso Y., 2011. Associations between dietary n-6 and n-3 fatty acids and arachidonic acid compositions in plasma and erythrocytes in young and elderly Japanese volunteers. Lipids Health Dis. 10, 138, https://doi.org/10.1186/1476-5....
22.
Krajewska-Bienias K.A., Czauderna M., Marounek M., Rozbicka-Wieczorek A.J., 2017. Diets containing selenized yeast, selenate, carnosic acid and fish oil change the content of fatty acids, tocopherols and cholesterol in the subcutaneous fat of lambs. J. Anim. Plant Sci. 27(6), 1781–1794, https://www.researchgate.net/p....
23.
Mertens D.R., 2002. Gravimetric determination of amylase-treated neutral detergent fiber in feeds with refluxing in beakers or crucibles: collaborative study. J. AOAC Int. 85(6), 1217–1240, PMID: 124771.
24.
Muhizi S., Kim I.H., 2021. Effect of dietary poppy (Papaver somniferum L.) seed meal supplementation on growth performance, nutrient digestibility, faecal microbiota and blood profile in growing-finishing pigs. J. Anim. Feed Sci. 30, 58–63, https://doi.org/10.22358/jafs/....
25.
Mužáková V., Meloun M., Jindrová A., Čegan A., 2019. The effect of fatty acids in red blood cell membranes on the dynamics of inflammatory markers following the coronary stent implantation. J. Pharm. Biomed. Anal. 166, 310–325, https://doi.org/10.1016/j.jpba....
26.
Niedźwiedzka M.K., Kowalczyk J., Czauderna M., 2008. Influence of selenate and linseed oil on fatty-acid and amino-acid profiles in the liver, muscles, fat tissues and blood plasma of sheep. J. Anim. Feed Sci. 17, 328–343, https://doi.org/10.22358/jafs/....
27.
National Research Council (NRC), 2012. Nutrient Requirements of Swine. 11th Edition, National Academies Press, Washington DC.
28.
Øverland M., Rørvik K.-A., Skrede A., 1999. High-fat diets improve the performance of growing–finishing pigs. Acta Agric. Scand., Sect. A, Animal Sci. 49, 83–88, https://doi.org/10.1080/090647....
29.
Pastorelli G., Serra V., Salvatori G., Redaelli V., Turin L., 2021. Evaluation of Boswellia serrata enriched diet on cytokine gene expression and reactive oxygen metabolites in weaning piglets. J. Anim. Feed Sci. 30, 119–127, https://doi.org/10.22358/jafs/....
30.
Poppitt S.D., Kilmartin P., Butler P., Keogh G.F., 2005. Assessment of erythrocyte phospholipid fatty acid composition as a biomarker for dietary MUFA, PUFA or saturated fatty acid intake in a controlled cross-over intervention trial. Lipids Health Dis. 4, 30, https://doi.org/10.1186/1476-5....
31.
Rozbicka-Wieczorek A.J., Więsyk E., Brzóska F., Śliwiński B., Kowalczyk J., Czauderna M., 2014. Efficiency of fatty acid accumulation into breast muscles of chickens fed diets with lycopene, fish oil and different chemical selenium forms. Afr. J. Biotechnol. 13, 1604–1613, https://doi.org/10.5897/AJB201....
32.
Shrivastava A., Gupta V.B., 2022. Methods for the determination of limit of detection and limit of quantitation of the analytical methods. Chron. Young Sci. 2, 21–25, https://doi.org/10.4103/2229-5....
33.
Skiba G., Poławska E., Raj S., Weremko D., Czauderna M., Wojtasik M., 2011. The influence of dietary fatty acids on their metabolism in liver and subcutaneous fat in growing pigs. J. Anim. Feed Sci. 20, 379–388, https://doi.org/10.22358/JAFS/....
34.
Sun Q., Ma J., Campos H., Hankinson S.E., Manson J.E., Stampfer M.J., Rexrode K.M., Willett W.C., Hu F.B., 2007. A prospective study of trans fatty acids in erythrocytes and risk of coronary heart disease. Circulation 115(14), 1858–1865, https://doi.org/10.1161/CIRCUL....
35.
Takkunen M., Agren J., Kuusisto J., Laakso M., Uusitupa M., Schwab U., 2013. Dietary fat in relation to erythrocyte fatty acid composition in men. Lipids 8(11), 1093–1102, https://doi.org/10.1007/s11745....
36.
Trivedi K., Le V., Nelson J.R., 2021. The case for adding eicosapentaenoic acid (icosapent ethyl) to the ABCs of cardiovascular disease prevention. Postgrad. Med. 133(1), 28–41, https://doi.org/10.1080/003254....
Share
RELATED ARTICLE
| ISSN: | 1230-1388 |
Assigning DOI numbers, introducing articles from supplements and recent publications to POL-index database, maintaining anti-plagiarism detection, electronic system to proceed manuscripts and webpage of the Journal with interactive pdf files, and language correction of manuscripts published in Journal of Animal and Feed Sciences are financed in the years 2018–2019 by the Ministry of Science and Higher Education from the funds for science popularization activities, Agreement No. 631/P-DUN/2018.
© 2006-2024 Journal hosting platform by Bentus
We process personal data collected when visiting the website. The function of obtaining information about users and their behavior is carried out by voluntarily entered information in forms and saving cookies in end devices. Data, including cookies, are used to provide services, improve the user experience and to analyze the traffic in accordance with the Privacy policy. Data are also collected and processed by Google Analytics tool (more).
You can change cookies settings in your browser. Restricted use of cookies in the browser configuration may affect some functionalities of the website.
You can change cookies settings in your browser. Restricted use of cookies in the browser configuration may affect some functionalities of the website.
