---
_id: '2864'
abstract:
- lang: eng
text: Using an electrospray tandem mass spectrometer as a concentration-sensitive
detector, a method has been developed to quantify femtomole amounts of plant growth
regulators (i.e. isoprenoid type cytokinins, zeatin, dihydrozeatin, isopentenyladenine
and their respective riboside and glucoside analogues) and the second messenger
adenosine 3':5'-cyclic monophosphate (3':5'-cAMP). Miniaturisation of the chromatographic
setup using capillary high performance liquid chromatographic (HPLC) ion spray
mass spectrometry increased the sensitivity to the low femtomole region. Application
of automated capillary column switching allowed the introduction of large injection
volumes into the HPLC system. Aliquots (25 μL) were injected into one dimension
of the HPLC set-up and stacked onto a micro pre-column. By means of mobile phase
switching the pre-column was back-flushed to introduce the analytes onto the analytical
column. For cytokinin analysis positive electrospray ionisation was used and resulted
in 2.5-25 fmol detection limits. Cyclic nucleotides were separated under ion-pair
conditions using tetrabutyl ammonium bromide as ion-pair reagent and were detected
under negative electrospray ionisation conditions. Here a 25 fmol detection limit
was determined. Following this approach, cytokinins and 3':5'-cAMP extracted from
only mg amounts of apical shoot meristem and chloroplasts obtained from Nicotiana
tabacum cv. Petit Havana SR1 were identified and quantified.
acknowledgement: The authors wish to thank J. Dupon for technical and logisticalassistance.
The authors also wish to thank the FWO-Vlaanderen/lotto (grant 32013394), the Flemish government (GOA-action) and the
Grant Agency of the Czech Republic (grant 206/96/K188) for financial support.
article_processing_charge: No
article_type: original
author:
- first_name: Erwin
full_name: Witters, Erwin
last_name: Witters
- first_name: Koen
full_name: Vanhoutte, Koen
last_name: Vanhoutte
- first_name: Walter
full_name: Dewitte, Walter
last_name: Dewitte
- first_name: Ivana
full_name: Macháčková, Ivana
last_name: Macháčková
- first_name: Eva
full_name: Benková, Eva
id: 38F4F166-F248-11E8-B48F-1D18A9856A87
last_name: Benková
orcid: 0000-0002-8510-9739
- first_name: Walter
full_name: Van Dongen, Walter
last_name: Van Dongen
- first_name: Eddy
full_name: Esmans, Eddy
last_name: Esmans
- first_name: Henri
full_name: Van Onckelen, Henri
last_name: Van Onckelen
citation:
ama: Witters E, Vanhoutte K, Dewitte W, et al. Analysis of cyclic nucleotides and
cytokinins in minute plant samples using phase system switching capillary electrospray
liquid chromatography tandem mass spectrometry. Phytochemical Analysis.
1999;10(3):143-151. doi:10.1002/(SICI)1099-1565(199905/06)10:3<143::AID-PCA441>3.0.CO;2-G
apa: Witters, E., Vanhoutte, K., Dewitte, W., Macháčková, I., Benková, E., Van Dongen,
W., … Van Onckelen, H. (1999). Analysis of cyclic nucleotides and cytokinins in
minute plant samples using phase system switching capillary electrospray liquid
chromatography tandem mass spectrometry. Phytochemical Analysis. Wiley-Blackwell.
https://doi.org/10.1002/(SICI)1099-1565(199905/06)10:3<143::AID-PCA441>3.0.CO;2-G
chicago: Witters, Erwin, Koen Vanhoutte, Walter Dewitte, Ivana Macháčková, Eva Benková,
Walter Van Dongen, Eddy Esmans, and Henri Van Onckelen. “Analysis of Cyclic Nucleotides
and Cytokinins in Minute Plant Samples Using Phase System Switching Capillary
Electrospray Liquid Chromatography Tandem Mass Spectrometry.” Phytochemical
Analysis. Wiley-Blackwell, 1999. https://doi.org/10.1002/(SICI)1099-1565(199905/06)10:3<143::AID-PCA441>3.0.CO;2-G.
ieee: E. Witters et al., “Analysis of cyclic nucleotides and cytokinins in
minute plant samples using phase system switching capillary electrospray liquid
chromatography tandem mass spectrometry,” Phytochemical Analysis, vol.
10, no. 3. Wiley-Blackwell, pp. 143–151, 1999.
ista: Witters E, Vanhoutte K, Dewitte W, Macháčková I, Benková E, Van Dongen W,
Esmans E, Van Onckelen H. 1999. Analysis of cyclic nucleotides and cytokinins
in minute plant samples using phase system switching capillary electrospray liquid
chromatography tandem mass spectrometry. Phytochemical Analysis. 10(3), 143–151.
mla: Witters, Erwin, et al. “Analysis of Cyclic Nucleotides and Cytokinins in Minute
Plant Samples Using Phase System Switching Capillary Electrospray Liquid Chromatography
Tandem Mass Spectrometry.” Phytochemical Analysis, vol. 10, no. 3, Wiley-Blackwell,
1999, pp. 143–51, doi:10.1002/(SICI)1099-1565(199905/06)10:3<143::AID-PCA441>3.0.CO;2-G.
short: E. Witters, K. Vanhoutte, W. Dewitte, I. Macháčková, E. Benková, W. Van Dongen,
E. Esmans, H. Van Onckelen, Phytochemical Analysis 10 (1999) 143–151.
date_created: 2018-12-11T12:00:00Z
date_published: 1999-05-01T00:00:00Z
date_updated: 2022-09-09T09:09:22Z
day: '01'
doi: 10.1002/(SICI)1099-1565(199905/06)10:3<143::AID-PCA441>3.0.CO;2-G
extern: '1'
intvolume: ' 10'
issue: '3'
language:
- iso: eng
month: '05'
oa_version: None
page: 143 - 151
publication: Phytochemical Analysis
publication_identifier:
issn:
- 0958-0344
publication_status: published
publisher: Wiley-Blackwell
publist_id: '3925'
quality_controlled: '1'
scopus_import: '1'
status: public
title: Analysis of cyclic nucleotides and cytokinins in minute plant samples using
phase system switching capillary electrospray liquid chromatography tandem mass
spectrometry
type: journal_article
user_id: ea97e931-d5af-11eb-85d4-e6957dddbf17
volume: 10
year: '1999'
...