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Fluorimetric determination of intra- and extracellular free amino acids in the microalgae Tetraselmis gracilis (Prasinophyceae) using monolithic column in reversed phase mode by José Carlo Pires Penteado; Marilda Rigobello-Masini; Cleber Wanderlei Liria; M. Terêsa Machini Miranda; Jorge Cesar Masini is a Chemistry article available to read on EtoBox.
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## Abstract This paper describes the development and application of an RP HPLC method using a C~18~ monolithic stationary phase for the separation and quantification of extra‐ and intracellular amino acids in a batch cultivation of the marine alga __Tetraselmis gracilis__. Fluorimetric detection was made after separation of the __o__‐phthaldialdehyde 2‐mercaptoethanol (OPA‐2MCE) derivatives using a binary gradient elution. Separation of 19 amino acids was achieved with resolution >1.5 in about 39 min at a flow rate of 1.5 mL/min. RSD of analyses in seawater medium ranged from 0.36% for Orn (0.50 μmol/L) to 12% for Ile (0.10 μmol/L). The main constituents of the intracellular dissolved free amino acids (DFAAs) in the exponential growth phase were arginine (Arg), asparagine (Asn), alanine (Ala), aspartic acid (Asp), glutamic acid (Glu), serine (Ser), glycine (Gly), glutamine (Gln), and leucine (Leu). The major amino acids excreted to the media were valine (Val), Ala, Ser, and Gly. The monolithic phase facilitates the analysis by shortening the separation time and saving solvents and instrumentation costs (indeed conventional HPLC instrumentation can be used, running at lower pressure
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- Author
- José Carlo Pires Penteado; Marilda Rigobello-Masini; Cleber Wanderlei Liria; M. Terêsa Machini Miranda; Jorge Cesar Masini
- Publisher
- John Wiley and Sons; Wiley (John Wiley & Sons); John Wiley & Sons Ltd.; Wiley (ISSN 1615-9306)
- Published
- 2009
- Language
- EN
- Field
- Chemistry (Physical Sciences)