Comparison of modified Matyash method to conventional solvent systems for polar metabolite and lipid extractions

Assessment of human plasma and urine sample preparation for reproducible and high-throughput UHPLC-MS clinical metabolic phenotyping

“…It needs to be reproducible, sensitive (to maximise metabolite and lipid extraction recovery) and should efficiently and rapidly denature proteins to halt residual enzymatic activity. [12][13][14] Methods which are high-throughput and lend themselves to automation provide opportunities to increase sample-throughput and the size of studies. Here we assess and subsequently recommend appropriate protocols of plasma (and serum by its association) and urine preparation for analysis by UHPLC-MS metabolomics (both sample types) and lipidomics (plasma only).…”

“…Such approaches can be: (i) monophasicthe addition of a water-miscible organic solvent to extract soluble compounds into a single phase; 15 and (ii) biphasicthe addition of immiscible organic solvent(s) and water (water provided by the biofluid and/or added water) to simultaneously extract hydrophilic metabolites and lipophilic metabolites into two separate phases. 13 Monophasic extracts can be immediately analysed (post-centrifugation) if the extraction solvents are compatible with UHPLC-MS. 12,15 For biphasic extractions 13 (and monophasic extractions using non-UHPLC-MS-compatible solvents 16 ) samples are dried and reconstituted in an appropriate solvent before analysis, thus increasing sample preparation complexity and time. The extraction solvent(s) influences metabolite/lipid solubility and protein removal efficiency 12,15 (sample stability is decreased if enzyme activity is not fully inactivated 14 ).…”

The rise of the nanomaterial metabolite corona, and emergence of the complete corona

“…35,88 There are a number of possible ways to do this, many of which have been applied in the metabolomics community to isolate metabolites from tissues or biofluids. [89][90][91] The most likely to work are successive washes in solvents ranging from the polar, such as water, to the less polar solvents such as methanol and hexane. 35 The chosen solvents would also be very much dependent upon the type of metabolite being isolated, for example the lipid corona will be more suited to an isopropanol or chloroform elution 92 whereas the more polar fractions would be better suited to water and/or methanol.…”

“…35 In cases where both the polar and non-polar Environmental Science: Nano Critical review portions of the corona need to be characterized then a biphasic extraction, such as the Bligh-Dyer, 93 Folch 94 and Matyash 95 methods or their derivatives, where phase separation occurs between water and chloroform/MTBE with polar metabolites in the aqueous phase and lipids in the organic phase is preferred. 91 These aforementioned methods also create a protein layer either at the solvent interface or at the bottom of the biphase, thus opening up the potential to be able to isolate the protein, polar metabolite and apolar metabolite portions of the complex corona. It is also likely that varying the pH of the solvent would have a significant effect on the elution of metabolites, either through hydrolysis of any bonds or changing the charge status of either the metabolite, NM or NM surface ligands thus releasing the metabolites.…”

Transient receptor potential canonical 5 (TRPC5) mediates inflammatory mechanical pain

“…Sample preparation. Lipids were extracted from samples as previously described (70). Samples were reconstituted in 250 µL Lipidyzer running buffer composed of dichloromethane:methanol with 10 mM ammonium acetate.…”