Optimization of a reversed-phase high performance liquid chromatography separation using an ion-pair reagent for the determination of carboxylic acids in plant materials

A method was developed to separate a number of mono-, di-and tri- carboxylic acids present in plant materials, using a C 18 column and a simple eluent containing a phosphate buffer and tetra-n-butylammonium phosphate, an ion-pairing agent. The effects of the ion-pairing agent, pH, presence or absenc...

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Bibliographic Details
Published inJournal of liquid chromatography & related technologies Vol. 20; no. 11; pp. 1773 - 1787
Main Authors Harrington, C.F, Roberts, D.J, Nickless, G
Format Journal Article
LanguageEnglish
Published Colchester Taylor & Francis Group 01.01.1997
Taylor & Francis
Subjects
Biological and medical sciences
chemical analysis
Chemical constitution
Fundamental and applied biological sciences. Psychology
high performance liquid chromatography
Plant physiology and development
Monocotyledones
Separation
Phase composition
Ion pair
HPLC chromatography
Mobile phase
Organic acids
Optimization
Reversed phase chromatography
Chemical modification
Gramineae
Carboxylic acid
Experimental design
Angiospermae
Plant origin
Agrostis capillaris
pH
Dicarboxylic acid
Spermatophyta
Festuca rubra
Factorial design
Chemometrics
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Summary:A method was developed to separate a number of mono-, di-and tri- carboxylic acids present in plant materials, using a C 18 column and a simple eluent containing a phosphate buffer and tetra-n-butylammonium phosphate, an ion-pairing agent. The effects of the ion-pairing agent, pH, presence or absence of organic modifier and the concentration of the buffer, on the chromatographic response function (CRF), were investigated using a two level factorial experimental design. The results indicated that a number of main, two and three factor effects were important. The largest effect was due to the interaction of pH with ion-pairing concentration and was further investigated using univariate experiments. The mobile phase developed consisted of 10 mM tetra-n-butylammonium phosphate and 20 mM potassium dihydrogen orthophosphate at pH 3.3. Formic, glutamic, quinic, succinic, malic and tartaric acids were extracted from 3 different grass ecotypes, the extracts were cleaned-up using a C 18 Sep-Pak® cartridge and then identified using the developed procedure.
ISSN:1082-6076
1520-572X
DOI:10.1080/10826079708006331