HPLC Determination of Amino Acids in Musts and Port Wine Using OPA/FMOC Derivatives

Abstract

Musts and port wines pose specific problems for amino acid analysis because of high sugar content. An HPLC technique was applied to the simultaneous quantification of primary and secondary amino acids in these matrices as their OPA-FMOC derivatives. This method enabled the fully automated analysis of 21 amino acids in 20 minutes. The average detection limit was 1.3 mmole/L, precision was 5%-12% for wine samples and accuracy by the standard addition method was 108.3 % (recovery value). Ten port wine and 43 must samples were analyzed for free amino acid contents. Proline represented 73% of the total for wine samples.

Analysis of free amino acids during fermentation by Bacillus subtilis using capillary electrophoresis

“…Conventional techniques for analysis of amino acids include ion exchange chromatography (IEC), using amino acid analyzer [9] and reverse-phase (RP)-HPLC [10,11], usually using a post-or pre-column derivatization with various kinds of chromophores [12] or fluorophores [13,14]. Amino acid analysis using an amino acid analyzer is expensive and time-consuming.…”

Analysis of Dabsyl-Cl Derivated Amino Acids by High Performance Liquid Chromatography and Tandem Mass Spectrometry

“…Recently, precolumn derivatization methods have become popular because they not only offer better chromatographic reproducibility (Cohen & Strydom, 1988), but also can be performed ubiquitously on any high performance liquid chromatographic (HPLC) system. Several reagents offering different advantages and feasibilities have been in common used: 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) (Pawlowska et al, 1993), 4-dimethylaminoazobenzene-4-sulfonyl chloride (Dabsyl-Cl) (Lin & Wang, 1980), dimethylaminonaphthalene-1-sulfonyl chloride (Dansyl-Cl) (Schmidt et al, 1979), 9-fluorenylmethyl chloroformate (FmocCl) (Einarsson et al, 1983), phenyl isothiocyanate (PITC) (Heinrikson & Meredith, 1984) and Ophthalaldehyde (OPA) (Herbert et al, 2000). However, neither a precolumn nor a postcolumn method has been successful in analyzing the free amino acids in a protein hydrolysate because all of the derivatization reagents mentioned above also react with peptides and proteins in the hydrolysate, thus creating interference (Gartenmann & Kochhar, 1999).…”

Isolation and Partial Characterization of Antagonistic Peptides Produced by Paenibacillus sp. Strain B2 Isolated from the Sorghum Mycorrhizosphere

“…A commercial sample of polymyxin B was hydrolyzed and analyzed in the same conditions. The analysis revealed that there were similar molar amounts of threonine, phenylalanine, and leucine in the antagonistic factor and polymyxin B. Diaminobutyric acid could not be accurately quantified using this technique since it gave two peaks, as previously observed in the case of ␥-aminobutyric acid (17), but the intensity of these two peaks was almost identical to that of the antagonistic factor and polymyxin B samples. Antagonistic activity of the Superdex-purified antagonistic factor.…”

“…Electrophoresis was performed in pyridine-acetic acid-water (1:5:94, vol/vol/vol) for 1 h at 300 V. Chromatography was performed in butanol-acetic acid-water-pyridine (4:1:2:1, vol/vol/vol/vol). Amino acids were visualized with ninhydrin, and the results were compared with the results obtained with samples of commercial polymyxin B or E. The same hydrolysate was used to quantify amino acids using precolumn derivatization with ortho-phthalaldehyde (OPA)/9-fluorenylmethyloxycarbonyl chloride (FMOC) and fluorescence detection (16,17).…”