Multi-response optimization of the extraction and derivatization protocol of selected polar metabolites from apple fruit tissue for GC-MS analysis.
Given the complexity of the plant metabolome exhibiting a wide spectrum of physico-chemical properties, finding the best compromise for GC-MS analysis is a challenging exercise. In this study, the GC-MS protocol for extracting and analyzing polar metabolites from apple tissue is optimized. We found pure methanol extraction to be slightly better as compared to the two phase methanol/chloroform/water extraction in terms of introducing less degradation of the extracted metabolites while being comparable in extraction efficiency and repeatability. The methanol extraction was superior to the single phase methanol/chloroform/water extraction in all performance measures. A multi-response optimization based on a desirability function was applied to optimize the derivatization. This procedure allows searching for optimal parameters while simultaneously considering overall detection enhancement of metabolites from various metabolic classes. A short oximation at a high temperature in combination with a low silylation temperature gave results similar to a longer oximation at low temperature in combination with a high silylation temperature. Increasing silylation time from 0.5 h to 2 h resulted in an improvement of the silylation reaction.
KU Leuven, Department of Biosystems (BIOSYST), Division of Mechatronics, Biostatistics and Sensors (MeBioS), Willem de Croylaan 42, bus 2428, B-3001, Leuven, Belgium.
Elias A
EA
Bekele
KU Leuven, Department of Biosystems (BIOSYST), Division of Mechatronics, Biostatistics and Sensors (MeBioS), Willem de Croylaan 42, bus 2428, B-3001, Leuven, Belgium.
Carlo E P
CE
Annaratone
KU Leuven, Department of Biosystems (BIOSYST), Division of Mechatronics, Biostatistics and Sensors (MeBioS), Willem de Croylaan 42, bus 2428, B-3001, Leuven, Belgium.
Maarten L A T M
ML
Hertog
KU Leuven, Department of Biosystems (BIOSYST), Division of Mechatronics, Biostatistics and Sensors (MeBioS), Willem de Croylaan 42, bus 2428, B-3001, Leuven, Belgium.
Bart M
BM
Nicolai
KU Leuven, Department of Biosystems (BIOSYST), Division of Mechatronics, Biostatistics and Sensors (MeBioS), Willem de Croylaan 42, bus 2428, B-3001, Leuven, Belgium. Electronic address: annemie.geeraerd@biw.kuleuven.be.
Annemie H
AH
Geeraerd
D005591:Chemical Fractionation / Q000379:methods*; D005638:Fruit / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D027845:Malus / Q000378:metabolism*; D010936:Plant Extracts / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D015203:Reproducibility of Results; D012997:Solvents / Q000737:chemistry
D010936:Plant Extracts; D012997:Solvents
Apple; Derivatization; Design of experiment; Gas chromatography–mass spectrometry; Metabolomics; Multi-response optimization