Braeburn' is an apple cultivar susceptible to the occurrence of internal browning (Braeburn Browning Disorder; BBD) during storage. This physiological disorder is characterised by the development of brown spots inside the fruit, eventually resulting in the formation of cavities. The objective of this study was to investigate the effects of the preharvest application of calcium, potassium and triazole fungicides on the postharvest 2 primary metabolites of 'Braeburn' fruit, and to offer a better understanding of the biochemical processes behind internal browning. The primary metabolites of 'Braeburn' cortex samples at harvest and after 2 weeks, weeks, 4 months and 8 months of storage at browning-inducing conditions were analysed using GC-MS. No significant difference in the primary metabolites was observed between the different levels of the applied preharvest applications. Early during storage, fruit developed browning, with the severity increasing with storage duration. This was correlated to a group of primary metabolites that showed either an increase (e.g., alanine, galactose, mannitol, sorbitol, xylose) or a decrease (e.g., malate, sucrose) in concentration with time. Radial distribution of the metabolites in the fruit tissue was also observed; some metabolites (e.g., galactose, mannitol) were higher in concentration in the inner cortex, while the concentrations of other metabolites (e.g., mannose, sucrose) were higher in the outer cortex.
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.
Belgian endive (Cichorium intybus L. var. foliosum Hegi), a popular produce in northern Europe, has been thoroughly studied in regard to its bitter sesquiterpene lactones content. Much less is known about on its sweetness and crunchiness, which are typically linked to the content of polar compounds such as sugars, organic acids and salts. Through HILIC-HPLC-MS, it was shown that simple sugars, amino acids, and potassium chloride are abundant in Belgian endive extracts. Subsequently, a HILIC-HPLC-ELSD method for the analysis of such compounds with run times below six minutes was developed. Recoveries varied between 80 and 110% and an average reproducibility was 7.5RSD%. Finally, the method was applied to the study of three difference Belgian endive varieties. Takine, a variety known for its sweet taste, was found to contain significantly higher levels of fructose, and lower levels of potassium and glutamine.
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