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A new method for quantitative determination of bitter components of Belgian endive, with a focus on sesquiterpene oxalates

ABSTRACT: The sesquiterpene lactones typical of the genera Cichorium and Lactuca were historically thought to be present in the free and glycosylated form, but recent developments have highlighted the presence of highly unusual oxalyl esters. In this work, first a liquid chromatography method that requires minimal sample preparation and can measure both free and conjugated sesquiterpene lactones was developed. Then the most common sample preparation techniques used in the past studies (e.g., solid-phase extraction, liquid/liquid extraction) were evaluated and shown to introduce significant bias against the polar oxalyl-terpenes. Finally the method was applied to a case study, highlighting the effect of variety, duration and temperature of forcing on the sesquiterpene lactone content of Belgian endive.

INSTRUMENT(S): QTof Micro (Micromass)

PROVIDER: MTBLS270 | MetaboLights | 2016-01-04

REPOSITORIES: MetaboLights

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Project description:Abstract: Wallerian degeneration (WD) is a process of autonomous distal degeneration of axons upon injury. Macrophages (MP) of the peripheral nervous system (PNS) are main cellular agent controlling this process. Some evidences suggest that resident PNS-MP along with MP of hematogenous origin may be involved but whether these two subsets exert distinct functions is unknown. Combining MP-designed fluorescent reporters mice, and coherent anti-stoke raman scattering (CARS) imaging of the sciatic nerve, we deciphered the spatio-temporal choreography of resident and recently recruited MP after injury and unveiled distinct functions of these subsets with recruited MP responsible of efficient myelin stripping and clearance while resident MP were involved in axonal regrowth. This work provides clues to tackle selectively cellular processes involved in neurodegenerative diseases. Methods: (relevant for this GEO dataset): RNAseq of sciatic nerve macrophages from mice after CCI: Sorted cells were lysed in 100µl of RA1/TCEP buffer (NucleoSpin RNA XS, Macherey-Nagel), snap frozen in liquid nitrogen and stored at -80C until RNA extraction. All samples were processed in parallel and RNA extraction (without Carrier RNA but with on-column DNase treatment) was performed according to manufacturer's instructions (NucleoSpin RNA XS, Macherey-Nagel). RNA was eluted in RNAse-free water. Preparation of cDNA libraries for RNAseq was done using the SmartSeq method according to manufacturer's instructions (SMART-Seq v4 Ultra Low Input RNA Kit for Sequencing, Clontech/TaKaRa). Due to the low number of cells, the total amount of eluted RNA was used as starting material for reverse transcription, followed by 18 cycles of pre-amplification. 1ng of cDNA were used for RNAseq sequencing library preparation, according to manufacturer's instructions (Nextera XT DNA Library Preparation, Illumina). Final samples pooled library prep were sequenced on a Nextseq 500 ILLUMINA with MidOutPut cartridge (2x130Millions of 75 bases reads) with 2 runs (4plex and 5plex), corresponding to 2x30Millions of (paired-end) reads per sample after demultiplexing.

			Action	DRS
	SG2W1001.CDF	Other
	SG2W101.CDF	Other
	SG2W201.CDF	Other
	SG2W301.CDF	Other
	SG2W401.CDF	Other

Dataset Information

A new method for quantitative determination of bitter components of Belgian endive, with a focus on sesquiterpene oxalates

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