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Current Metabolomics and Systems Biology (Discontinued)

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ISSN (Print): 2666-3384
ISSN (Online): 2666-3392

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TECHNICAL NOTE

A Robust Protocol for Extracting Aqueous Metabolites of High Lipid Sera

Author(s): Matthew C. Taddeo, Emma J. Robinson, Noy Y. Hassid, Xin Chu, Weixing Shi, Craig Wood, Christopher Still and David Rovnyak*

Volume 7, Issue 1, 2020

Page: [67 - 72] Pages: 6

DOI: 10.2174/2213235X07666190124120112

Price: $65

Abstract

Background: With the increasing focus of metabolomic methods on obesityrelated diseases, it is important to consider how sample handling may need to be adapted for the high compositions of lipids that can occur in such subjects.

Introduction: High-lipid (cloudy, milky appearances; a.k.a. lipemic) biofluids are common in very high BMI subjects. Organic extractions of biofluids are useful for removing protein backgrounds, inactivating capsid viruses, and yielding relatively stable samples with excellent spectroscopic characteristics. This work considered how acetonitrile extractions, which are widely used, perform on lipemic sera.

Results: In this technical note, we report the observation and remediation of a liquid-liquid phase separation in acetonitrile extractions of many lipemic sera. This unexpected behavior can be challenging to identify, especially if working with small volumes. The liquid-liquid separation shows a high miscibility of proteins in both liquid phases that impairs NMR data quality. We also report a simple temperature-based adaption of the acetonitrile extraction procedure that consistently results in a single aqueous phase and eliminates unwanted constituents.

Conclusion: A robust approach to achieving reproducible, high quality samples of aqueous metabolites from lipemic sera from very high BMI subjects should be of utility in expanding metabolomics applications to lipemic biofluids.

Keywords: Acetonitrile, organic, extraction, lipemic, BMI, NMR, phase separation.

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[1]
Wolak-Dinsmore, J.; Gruppen, E.G.; Shalaurova, I.; Matyus, S.P.; Grant, R.P.; Gegen, R.; Bakker, S.J.L.; Otvos, J.D.; Connelly, M.A.; Dullaart, R.P.F. A novel NMR-based assay to measure circulating concentrations of branched-chain amino acids: Elevation in subjects with type 2 diabetes mellitus and association with carotid intima media thickness. Clin. Biochem., 2018, 54, 92-99.
[http://dx.doi.org/10.1016/j.clinbiochem.2018.02.001] [PMID: 29432757]
[2]
Markgraf, D.F.; Al-Hasani, H.; Lehr, S. Lipidomics-Reshaping the Analysis and Perception of Type 2 Diabetes. Int. J. Mol. Sci., 2016, 17(11), 17.
[http://dx.doi.org/10.3390/ijms17111841] [PMID: 27827927]
[3]
Klein, M.S.; Shearer, J. Metabolomics and Type 2 Diabetes: Translating Basic Research into Clinical Application. J. Diabetes Res., 2016, 20163898502
[http://dx.doi.org/10.1155/2016/3898502] [PMID: 26636104]
[4]
Newgard, C.B. Metabolomics and Metabolic Diseases: Where Do We Stand? Cell Metab., 2017, 25(1), 43-56.
[http://dx.doi.org/10.1016/j.cmet.2016.09.018] [PMID: 28094011]
[5]
Bain, J.R.; Stevens, R.D.; Wenner, B.R.; Ilkayeva, O.; Muoio, D.M.; Newgard, C.B. Metabolomics applied to diabetes research: moving from information to knowledge. Diabetes, 2009, 58(11), 2429-2443.
[http://dx.doi.org/10.2337/db09-0580] [PMID: 19875619]
[6]
Gonzalez-Franquesa, A.; Burkart, A.M.; Isganaitis, E.; Patti, M.E. What Have Metabolomics Approaches Taught Us About Type 2 Diabetes? Curr. Diab. Rep., 2016, 16(8), 74.
[http://dx.doi.org/10.1007/s11892-016-0763-1] [PMID: 27319324]
[7]
Bloomgarden, Z. Diabetes and branched-chain amino acids: What is the link? J. Diabetes, 2018, 10(5), 350-352.
[http://dx.doi.org/10.1111/1753-0407.12645] [PMID: 29369529]
[8]
Wang, T.J.; Larson, M.G.; Vasan, R.S.; Cheng, S.; Rhee, E.P.; McCabe, E.; Lewis, G.D.; Fox, C.S.; Jacques, P.F.; Fernandez, C.; O’Donnell, C.J.; Carr, S.A.; Mootha, V.K.; Florez, J.C.; Souza, A.; Melander, O.; Clish, C.B.; Gerszten, R.E. Metabolite profiles and the risk of developing diabetes. Nat. Med., 2011, 17(4), 448-453.
[http://dx.doi.org/10.1038/nm.2307] [PMID: 21423183]
[9]
Miele, M.M.; Irving, B.A.; Wenrich, B.R.; Martin, P.L. Reproducibility and Stability of Aqueous Metabo-lite Levels in Extracted Serum by NMR Spectroscopy. Curr. Metabolomics, 2017, 5, 45-54.
[http://dx.doi.org/10.2174/2213235X04666160711160340]
[10]
Nagana Gowda, G.A.; Gowda, Y.N.; Raftery, D. Expanding the limits of human blood metabolite quantitation using NMR spectroscopy. Anal. Chem., 2015, 87(1), 706-715.
[http://dx.doi.org/10.1021/ac503651e] [PMID: 25485990]
[11]
Mckay, R.T. How the 1D-NOESY suppresses solvent signal in metabonomics NMR spectroscopy: An ex-amination of the pulse sequence components and evolution. Concepts Magn. Reson., 2011, 38A, 197-220.
[http://dx.doi.org/10.1002/cmr.a.20223]
[12]
Faria, R.P.V.; Pereira, C.S.M. Silva, Viviana M. T. M.; Loureiro, J. M.; Rodrigues, A. E. Glycerol valori-sation as biofuels: Selection of a suitable solvent for an innovative process for the synthesis of GEA. Chem. Eng. J., 2013, 233, 159-167.
[http://dx.doi.org/10.1016/j.cej.2013.08.035]
[13]
Dhamole, P.B.; Mahajan, P.; Feng, H. Phase Separa-tion Conditions for Sugaring-Out in Acetoni-trile−Water Systems. J. Chem. Eng. Data, 2010, 55, 3803-3806.
[http://dx.doi.org/10.1021/je1003115]
[14]
FUJINAGA T SHUHARA A HORI T. Character-istic properties in phase-separation of acetoni-trile/water/chloroform ternary solvent system and their analytical application; Study of solvent syner-gism for the selective extraction. IV. Bunseki Kagaku, 1984, 33, 159-164.
[http://dx.doi.org/10.2116/bunsekikagaku.33.3_159]
[15]
DelMastro, T.; Snow, N.H.; Murphy, W.R.; Sowa, J.R. Polyol-induced partitioning of essential oils in wa-ter/acetonitrile solvent mixtures. J. Liq. Chromatogr. Relat. Technol., 2017, 40, 376-383.
[http://dx.doi.org/10.1080/10826076.2017.1308379]
[16]
Taha, M.; Teng, H.; Lee, M. Phase diagrams of ace-tonitrile or (acetone+water+EPPS) buffer phase sepa-ration systems at 298.15K and quantum chemical modeling. J. Chem. Thermodyn., 2012, 54, 134-141.
[http://dx.doi.org/10.1016/j.jct.2012.03.026]
[17]
Gu, T.; Gu, Y.; Zheng, Y.; Wiehl, P.E.; Kopchick, J.J. Phase separation of acetonitrile-water mixture in pro-tein purification. Separations Technology, 1994, 4, 258-260.
[http://dx.doi.org/10.1016/0956-9618(94)80031-6]
[18]
Alum, M.F.; Shaw, P.A.; Sweatman, B.C.; Ubhi, B.K.; Haselden, J.N.; Connor, S.C. 4,4-Dimethyl-4-silapentane-1-ammonium trifluoroacetate (DSA), a promising universal internal standard for NMR-based metabolic profiling studies of biofluids, including blood plasma and serum. Metabolomics, 2008, 4, 122-127.
[http://dx.doi.org/10.1007/s11306-008-0103-9]
[19]
Shimizu, A.; Ikeguchi, M.; Sugai, S. Appropriateness of DSS and TSP as internal references for (1)H NMR studies of molten globule proteins in aqueous media. J. Biomol. NMR, 1994, 4(6), 859-862.
[http://dx.doi.org/10.1007/BF00398414] [PMID: 22911388]
[20]
Nagana Gowda, G.A.; Raftery, D. Can NMR solve some significant challenges in metabolomics? J. Magn. Reson., 2015, 260, 144-160.
[http://dx.doi.org/10.1016/j.jmr.2015.07.014] [PMID: 26476597]

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