Solid Phase Microextraction Sampling: A Probe-based Technique for Sample Extraction from Graft Tissues

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

1. 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 is recommended per time point).
   NOTE: The length and type of extraction phase may be adjusted based on the mode of study, the p....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
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