Multi-response optimization of the extraction and derivatization protocol of selected polar metabolites from apple fruit tissue for GC–MS analysis

Abstract

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.

Dynamic Labeling Reveals Temporal Changes in Carbon Re-Allocation within the Central Metabolism of Developing Apple Fruit

“…Extraction and derivatization of polar metabolites were performed following the protocols described by Bekele et al ( 2014 ). The frozen hypanthium tissue samples were homogenized using Mixer Mill (Retsch, MM 200, Haan, Germany) at a frequency of 30 Hz for 1 min.…”

Investigation of Organic Acids in Saffron Stigmas (Crocus sativus L.) Extract by Derivatization Method and Determination by GC/MS

“…As the reaction time was prolonged, the signal response of organic acid derivatives remained constant. Similarly, Elias and co-authors found that long term silylation-derivatization process was beneficial to stearic acid and glucose-6-phosphate [ 25 ]. Therefore, the derivatization reaction between MTBSTFA and organic acids was carried out for 90 min at 130 °C.…”

“…This is in line with the findings of Koek, who showed that organic acids and sugars need relatively low volumes of the silylating agent [ 27 ]. It could be explained as molecular interaction because the reactivity of TMS groups is low to oxygen in organic acids, having a lower number of unshared electrons, higher steric hindrance, and transition state energy [ 25 ].…”

Carboxylated graphene oxide/polyvinyl chloride as solid-phase extraction sorbent combined with ion chromatography for the determination of sulfonamides in cosmetics

“…Response surface methodology (RSM) including the Box-Behnken design (BBD) and central composite design (CCD), is a mathematical and statistical tool for assessing the effect of variables on a response, which is the preferred way to find the best compromising set of parameters in sample preparation [31]. A number of algorithms have been developed, such as Khuri-Conlon approach, quality loss function and desirability function [32]. Among them, the desirability function is the most widely used methodology for simultaneously optimizing multiple variables.…”