Microfluidic capillary zone electrophoresis mass spectrometry analysis of alkaloids in Lobelia cardinalis transgenic and mutant plant cell cultures

Zachary D. Kelley; D. Trent Rogers; John M. Littleton; Bert C. Lynn

doi:10.1002/elps.201900220

Microfluidic capillary zone electrophoresis mass spectrometry analysis of alkaloids in Lobelia cardinalis transgenic and mutant plant cell cultures

Zachary D. Kelley, D. Trent Rogers, John M. Littleton, Bert C. Lynn

Research output: Contribution to journal › Article › peer-review

13 Scopus citations

Abstract

Application of a microfluidic CE^* device for CZE-MS allows for fast, rapid, and in-depth analysis of large sample sets. This microfluidic CZE-MS device, the 908 Devices ZipChip, involves minimal sample preparation and is ideal for small cation analytes, such as alkaloids. Here, we evaluated the microfluidic device for the analysis of alkaloids from Lobelia cardinalis hairy root cultures. Extracts from wild-type, transgenic, and selected mutant plant cultures were analyzed and data batch processed using the mass spectral processing software MZmine2 and the statistical software Prism 8. In total 139 features were detected as baseline resolved peaks via the MZmine2 software optimized for the electrophoretic separations. Statistically significant differences in the relative abundance of the primary alkaloid lobinaline (C₂₇H₃₄N₂), along with several putative “lobinaline-like” molecules were observed utilizing this approach. Additionally, a method for performing both targeted and untargeted MS/MS experiments using the microfluidic device was developed and evaluated. Coupling data-processing software with CZE-MS data acquisition has enabled comprehensive metabolomic profiles from plant cell cultures to be constructed within a single working day.

Original language	English
Pages (from-to)	2921-2928
Number of pages	8
Journal	Electrophoresis
Volume	40
Issue number	22
DOIs	https://doi.org/10.1002/elps.201900220
State	Published - Nov 1 2019

Bibliographical note

Funding Information:
The authors gratefully acknowledge support from the National Institute on Alcohol Abuse and Alcoholism of the National Institutes of Health under the award number R44AA025804. Additionally, ZDK and BCL acknowledge the UK Office of Research for funding the acquisition of the ZipChip equipment.

Funding Information:
The authors gratefully acknowledge support from the National Institute on Alcohol Abuse and Alcoholism of the National Institutes of Health under the award number R44AA025804. Additionally, ZDK and BCL acknowledge the UK Office of Research for funding the acquisition of the ZipChip equipment .

Publisher Copyright:
© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

Keywords

Alkaloid
Bioinformatics
Microfluidic
Tandem MS

ASJC Scopus subject areas

Analytical Chemistry
Biochemistry
Clinical Biochemistry

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10.1002/elps.201900220

Cite this

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title = "Microfluidic capillary zone electrophoresis mass spectrometry analysis of alkaloids in Lobelia cardinalis transgenic and mutant plant cell cultures",

abstract = "Application of a microfluidic CE* device for CZE-MS allows for fast, rapid, and in-depth analysis of large sample sets. This microfluidic CZE-MS device, the 908 Devices ZipChip, involves minimal sample preparation and is ideal for small cation analytes, such as alkaloids. Here, we evaluated the microfluidic device for the analysis of alkaloids from Lobelia cardinalis hairy root cultures. Extracts from wild-type, transgenic, and selected mutant plant cultures were analyzed and data batch processed using the mass spectral processing software MZmine2 and the statistical software Prism 8. In total 139 features were detected as baseline resolved peaks via the MZmine2 software optimized for the electrophoretic separations. Statistically significant differences in the relative abundance of the primary alkaloid lobinaline (C27H34N2), along with several putative “lobinaline-like” molecules were observed utilizing this approach. Additionally, a method for performing both targeted and untargeted MS/MS experiments using the microfluidic device was developed and evaluated. Coupling data-processing software with CZE-MS data acquisition has enabled comprehensive metabolomic profiles from plant cell cultures to be constructed within a single working day.",

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author = "Kelley, {Zachary D.} and Rogers, {D. Trent} and Littleton, {John M.} and Lynn, {Bert C.}",

note = "Funding Information: The authors gratefully acknowledge support from the National Institute on Alcohol Abuse and Alcoholism of the National Institutes of Health under the award number R44AA025804. Additionally, ZDK and BCL acknowledge the UK Office of Research for funding the acquisition of the ZipChip equipment. Funding Information: The authors gratefully acknowledge support from the National Institute on Alcohol Abuse and Alcoholism of the National Institutes of Health under the award number R44AA025804. Additionally, ZDK and BCL acknowledge the UK Office of Research for funding the acquisition of the ZipChip equipment . Publisher Copyright: {\textcopyright} 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim",

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N1 - Funding Information: The authors gratefully acknowledge support from the National Institute on Alcohol Abuse and Alcoholism of the National Institutes of Health under the award number R44AA025804. Additionally, ZDK and BCL acknowledge the UK Office of Research for funding the acquisition of the ZipChip equipment. Funding Information: The authors gratefully acknowledge support from the National Institute on Alcohol Abuse and Alcoholism of the National Institutes of Health under the award number R44AA025804. Additionally, ZDK and BCL acknowledge the UK Office of Research for funding the acquisition of the ZipChip equipment . Publisher Copyright: © 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

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N2 - Application of a microfluidic CE* device for CZE-MS allows for fast, rapid, and in-depth analysis of large sample sets. This microfluidic CZE-MS device, the 908 Devices ZipChip, involves minimal sample preparation and is ideal for small cation analytes, such as alkaloids. Here, we evaluated the microfluidic device for the analysis of alkaloids from Lobelia cardinalis hairy root cultures. Extracts from wild-type, transgenic, and selected mutant plant cultures were analyzed and data batch processed using the mass spectral processing software MZmine2 and the statistical software Prism 8. In total 139 features were detected as baseline resolved peaks via the MZmine2 software optimized for the electrophoretic separations. Statistically significant differences in the relative abundance of the primary alkaloid lobinaline (C27H34N2), along with several putative “lobinaline-like” molecules were observed utilizing this approach. Additionally, a method for performing both targeted and untargeted MS/MS experiments using the microfluidic device was developed and evaluated. Coupling data-processing software with CZE-MS data acquisition has enabled comprehensive metabolomic profiles from plant cell cultures to be constructed within a single working day.

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Microfluidic capillary zone electrophoresis mass spectrometry analysis of alkaloids in Lobelia cardinalis transgenic and mutant plant cell cultures

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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