Microchip analysis of plant glucosinolates

Abstract

We describe a new and selective analytical method for the separation and quantitation of plant glucosinolates. The new method, which utilizes microchip CE (micro-CE) with fluorescence detection, circumvents the multistep procedures characteristic of conventional methods. Glucosinolates form charge transfer complexes with the xanthene dyes phloxine-B and eosin-B. The glucosinolates-phloxine-B complex cannot be excited at 470 nm. Thus, the decrease in peak intensity of phloxine-B after complex formation is used to quantitatively measure total glucosinolates in Arabidopsis thaliana seeds. For qualitative analysis, complex formation with eosin-B is used. The sensitivity of eosin-B detection at excitation/emission 470 nm/540 nm was low. However, sensitivity increased following complex formation with sinigrin (> or =3 microg/mL). A batch-learning, self-organizing map was applied to visualize and organize analytical data into 2-D matrix with similar and related data clustered together or near each other. This organized matrix was used to optimize electrophoretic conditions for the analysis. This study suggests potential applications of micro-CE in plant metabolomics analyses without use of labeling fluorophores.

Arabidopsis thaliana and omics approaches: a review

“…In recent years, the rapid progress in the determination, quanti cation, identi cation and comprehension of proteins has been possible due to the use of model organisms such as A. thaliana. Improvements in the techniques for proteomics, including plant proteomics, based on existing platforms such as 2-DE, 2-LC and MS and some new techniques, including tandem a nity and protein chips, have been observed [38][39][40][41][42][43][44][45][46][47]. e availability of the entire genomic sequence of A. thaliana provides unique opportunities for the use of a postgenomic tool such as proteomics in its full capacity [11,[17][18][19][20][21][22]48,49].…”

CE‐MS for metabolomics: Developments and applications in the period 2008–2010

“…More recently, microchip CE‐LIF was used for the profiling of glucosinolates in extracts of Arabidopsis thaliana seeds. 17. Electrophoretic separation of ten different glucosinolates was performed in <3 min (Fig.…”

“…Ten different glucosinolates were detected assuming that no co‐migration of components occurred. Reproduced from 17 with permission.…”

Recent developments in instrumentation for capillary electrophoresis and microchip‐capillary electrophoresis

“…Although other zones of the spectra such as the infrared spectral region have been used [56], UV-Vis is the default detection method in most commercial instruments. Pertinent examples include Beckman [57], Shimadzu [58], Waters [59], Hitachi [60], Caliper [61], Agilent [62, 63], or Bio-Rad [64], just to name a few. The advantages of UV-Vis detection include versatility (most organic analytes absorb light in this range), speed, and a wide variety of commercial sources.…”