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Amino acid analysis by micellar electrokinetic chromatography with laser-induced fluorescence detection: application to nanolitre-volume biological samples from Arabidopsis thaliana and Myzus persicae.
Electrophoresis. 2005 Feb; 26(4-5):911-9.E

Abstract

Amino acids were derivatised with 4-fluoro-7-nitrobenzo-2,1,3-oxadiazol (NBD-F), separated by micellar electrokinetic chromatography (MEKC), and detected by argon-ion (488 nm) laser-induced fluorescence. The optimised MEKC background electrolyte conditions were: 40 mM sodium cholate, 5 mM beta-cyclodextrin in 20 mM aqueous borate buffer, pH 9.1, with 7% v/v acetonitrile. Using these conditions, 19 amino acids were separated within 17 min. The limits of detection were in the range of 7.6-42.2 pmol/mL and limits of quantitation from 0.05-0.14 nmol/mL. The method was systematically validated for injection volume error, migration time variation, calibration linearity, accuracy, precision, and recovery. Nanolitre volume samples of phloem sap of individual sieve element cells from the plant Arabidopsis thaliana and honeydew from the aphid Myzus persicae were directly analysed with this method. Quantitative amino acid concentrations in these two biological matrices were profiled for the first time. This method is particularly important because it allows the complete profile of the amino acids obtained from individual phloem elements, allowing cell to cell and plant to plant variation to be quantified, which to date has not been possible with Arabidopsis thaliana.

Authors+Show Affiliations

Centre for Analytical Bioscience, School of Pharmacy, University of Nottingham, Nottingham, UK.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Validation Study

Language

eng

PubMed ID

15714547

Citation

Zhu, Xunlin, et al. "Amino Acid Analysis By Micellar Electrokinetic Chromatography With Laser-induced Fluorescence Detection: Application to Nanolitre-volume Biological Samples From Arabidopsis Thaliana and Myzus Persicae." Electrophoresis, vol. 26, no. 4-5, 2005, pp. 911-9.
Zhu X, Shaw PN, Pritchard J, et al. Amino acid analysis by micellar electrokinetic chromatography with laser-induced fluorescence detection: application to nanolitre-volume biological samples from Arabidopsis thaliana and Myzus persicae. Electrophoresis. 2005;26(4-5):911-9.
Zhu, X., Shaw, P. N., Pritchard, J., Newbury, J., Hunt, E. J., & Barrett, D. A. (2005). Amino acid analysis by micellar electrokinetic chromatography with laser-induced fluorescence detection: application to nanolitre-volume biological samples from Arabidopsis thaliana and Myzus persicae. Electrophoresis, 26(4-5), 911-9.
Zhu X, et al. Amino Acid Analysis By Micellar Electrokinetic Chromatography With Laser-induced Fluorescence Detection: Application to Nanolitre-volume Biological Samples From Arabidopsis Thaliana and Myzus Persicae. Electrophoresis. 2005;26(4-5):911-9. PubMed PMID: 15714547.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Amino acid analysis by micellar electrokinetic chromatography with laser-induced fluorescence detection: application to nanolitre-volume biological samples from Arabidopsis thaliana and Myzus persicae. AU - Zhu,Xunlin, AU - Shaw,P Nicholas, AU - Pritchard,Jeremy, AU - Newbury,John, AU - Hunt,Emma J, AU - Barrett,David A, PY - 2005/2/17/pubmed PY - 2005/7/27/medline PY - 2005/2/17/entrez SP - 911 EP - 9 JF - Electrophoresis JO - Electrophoresis VL - 26 IS - 4-5 N2 - Amino acids were derivatised with 4-fluoro-7-nitrobenzo-2,1,3-oxadiazol (NBD-F), separated by micellar electrokinetic chromatography (MEKC), and detected by argon-ion (488 nm) laser-induced fluorescence. The optimised MEKC background electrolyte conditions were: 40 mM sodium cholate, 5 mM beta-cyclodextrin in 20 mM aqueous borate buffer, pH 9.1, with 7% v/v acetonitrile. Using these conditions, 19 amino acids were separated within 17 min. The limits of detection were in the range of 7.6-42.2 pmol/mL and limits of quantitation from 0.05-0.14 nmol/mL. The method was systematically validated for injection volume error, migration time variation, calibration linearity, accuracy, precision, and recovery. Nanolitre volume samples of phloem sap of individual sieve element cells from the plant Arabidopsis thaliana and honeydew from the aphid Myzus persicae were directly analysed with this method. Quantitative amino acid concentrations in these two biological matrices were profiled for the first time. This method is particularly important because it allows the complete profile of the amino acids obtained from individual phloem elements, allowing cell to cell and plant to plant variation to be quantified, which to date has not been possible with Arabidopsis thaliana. SN - 0173-0835 UR - https://www.unboundmedicine.com/medline/citation/15714547/Amino_acid_analysis_by_micellar_electrokinetic_chromatography_with_laser_induced_fluorescence_detection:_application_to_nanolitre_volume_biological_samples_from_Arabidopsis_thaliana_and_Myzus_persicae_ DB - PRIME DP - Unbound Medicine ER -