Solid Phase Microextraction Sampling: A Probe-based Technique for Sample Extraction from Graft Tissues
Published: April 30, 2023
Abstract
Source: Stryjak, I. et al. Using a Chemical Biopsy for Graft Quality Assessment. J. Vis. Exp. (2020)
In this video, we demonstrate SPME-probe-based extraction of metabolites from a pre-harvested donor kidney. The extracted metabolites can be used to perform metabolomic and lipidomic analysis in order to assess the organ quality for transplantation.
Protocol
All procedures involving human participants have been performed in compliance with the institutional, national and international guidelines for human welfare and have been reviewed by the local institutional review board. 1. Preparation of Probes Prepare titanium-nickel alloy probes (40 mm length; 0.2 mm diameter) coated with 7 mm mix-mode sorbent. The number of probes depends on the time points targeted during the entire procedure and the number of replicates (3 i…
Divulgaciones
The authors have nothing to disclose.
Materials
| Acetic acid | Merck | 5330010050 | Mobile phase additive |
| Acetonitrile | Alchem | 696-34967-4X2.5L | HPLC solvent |
| Ammonium acetate | Merck | 5330040050 | Mobile phase additive |
| BENCHMIXER XL MULTI-TUBE VORTEXER | Benchmark Scientific | BV1010 | Vortex mixer |
| Caps | Perlan Technologies | 5183-2076 | Blue scrw tp, pre-slit PTFE/Si spta, 100PK |
| Chloroform | Merck | 1024441000 | |
| Discovery HS F5 Supelguard Cartridge, 3 μm, L × I.D. 2 cm × 2.1 mm | Merck | 567570-U | HPLC guard column |
| Discovery HS F5, 2.1 mm x 100 mm, 3 μm | Merck | 567502-U | HPLC column |
| Formic acid | Alchem | 497-94318-50ML | Mobile phase additive |
| Glass vials | Perlan Technologies | 5182-0714 | |
| HILIC Luna 3 μm, 200A, 100 x 2.0 mm | Shim-Pol | PHX-00D-4449-B0 | HPLC column |
| HILIC SecurityGuard Cartridge, 3 μm, 4 x 2.0 mm | Shim-Pol | PHX-AJ0-8328 | HPLC guard column |
| Isopropanol | Alchem | 231-AL03262500 | HPLC solvent |
| Methanol | Alchem | 696-34966-4X2.5L | HPLC solvent |
| Nano-pure water | Merck | 1037281002 | HPLC solvent |
| Q Exactive Focus hybrid quadrupole-Orbitrap MS | Thermo Scientific | Q Exactive Focus | Mass Spectrometer |
| SeQuant ZIC-cHILIC 3µm,100Å 100 x 2.1 mm | Merck | 1506570001 | HPLC column |
| SeQuant ZIC-HILIC Guard Kit 20 x 2.1 mm | Merck | 1504360001 | HPLC guard column |
| SPME LC fiber probes, mixed mode | Supelco | prototype fibers | |
| UltiMate 3000 HPLC systems | Thermo Scientific | UltiMate 3000 | HPLC system |
| Vial inserts (deactivated) | Perlan Technologies | 5181-8872 | |
| XSelect CSH C18 3.5μm 2.1x75mm | Waters | 186005644 | HPLC column |
| XSelect CSH C18 VanGuard Cartridge 3.5μm, 2.1x5mm | Waters | 186007811 | HPLC guard column |
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Solid Phase Microextraction Sampling: A Probe-based Technique for Sample Extraction from Graft Tissues. J. Vis. Exp. (Pending Publication), e20554, doi: (2023).