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Simultaneous separation and quantitation of amino acids and polyamines of forest tree tissues and cell cultures within a single high-performance liquid chromatography run using dansyl derivatizationAuthor(s): Rakesh Minocha; Stephanie Long
Source: Journal of Chromatography A. 1035: 63-73.
Publication Series: Scientific Journal (JRNL)
Station: Northern Research Station
PDF: Download Publication (623.29 KB)
DescriptionThe objective of the present study was to develop a rapid HPLC method for simultaneous separation and quantitation of dansylated amino acids and common polyamines in the same matrix for analyzing forest tree tissues and cell cultures. The major modifications incorporated into this method as compared to previously published HPLC methods for separation of only dansyl amino acids include: use of a l0 cm column to reduce the total run time by approximately 15 min; modification of the dansyl derivatization process and gradient profile to elute amino acids and common polyamines within the same run; addition of steps for column cleaning within each run; shorter re-equilibration time; and finally, column cleaning and physically reversing the column at the end of a loop of samples. These changes improved peak resolution and increased column longevity by several-fold. Over 1000 foliar samples from mature forest trees could be analyzed with the same column as compared to only 200-250 samples before the incorporation of these changes. This method eluted 22 amino acids within 40 min plus all three common polyamines between 44 and 47 min. The total run time is 53.6 min for amino acids only and 55.6 min for both amino acids and polyamines.
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CitationMinocha, Rakesh; Long, Stephanie. 2004. Simultaneous separation and quantitation of amino acids and polyamines of forest tree tissues and cell cultures within a single high-performance liquid chromatography run using dansyl derivatization. Journal of Chromatography A. 1035: 63-73.
KeywordsDerivatization, LC, Amino acids, Dns derivatives, Polyamines, Amines, Putrescine, Spermidine, Spermine
- Separation and quantification of monothiols and phytochelatins from a wide variety of cell cultures and tissues of trees and other plants using high performance liquid chromatography
- High-performance liquid chromatographic method for the determination of dansyl-polyamines
- Extraction and estimation of the quantity of calcium oxalate crystals in the foliage of conifer and hardwood trees
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