Optimised Method for Short-Chain Fatty Acid Profiling of Bovine Milk and Serum |
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| Authors: | Cheng Li Zhiqian Liu Carolyn Bath Leah Marett Jennie Pryce Simone Rochfort |
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| Institution: | 1.Agriculture Victoria Research, AgriBio, 5 Ring Road, Bundoora, VIC 3083, Australia; (C.L.); (C.B.); (J.P.); (S.R.);2.School of Applied Systems Biology, La Trobe University, Bundoora, VIC 3083, Australia;3.Agriculture Victoria Research, Ellinbank Centre, Ellinbank, VIC 3821, Australia;4.Centre for Agricultural Innovation, School of Agriculture and Food, Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Melbourne, VIC 3010, Australia |
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| Abstract: | Short-chain fatty acids (SCFA, C2-C5) in milk and serum are derived from rumen bacterial fermentation and, thus, have the potential to be used as biomarkers for the health status of dairy cows. Currently, there is no comprehensive and validated method that can be used to analyse all SCFAs in both bovine serum and milk. This paper reports an optimised protocol, combining 3-nitrophenylhydrazine (3-NPH) derivatisation and liquid chromatography-mass spectrometry (LC-MS) analysis for quantification of SCFA and β-hydroxybutyric acid (BHBA) in both bovine milk and bovine serum. This method is sensitive (limit of detection (LOD) ≤ 0.1 µmol/L of bovine milk and serum), accurate (recovery 84–115% for most analytes) and reproducible (relative standard deviation (RSD) for repeated analyses below 7% for most measurements) with a short sample preparation step. The application of this method to samples collected from a small cohort of animals allowed us to reveal a large variation in SCFA concentration between serum and milk and across different animals as well as the strong correlation of some SCFAs between milk and serum samples. |
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| Keywords: | bovine milk serum short-chain fatty acids liquid chromatography-mass spectrometry |
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