Rapid determination of amino acids in biological samples using a monolithic silica column

Song, Yanting; Funatsu, Takashi; Tsunoda, Makoto

doi:10.1007/s00726-011-0914-2

Cited by 29 publications

(19 citation statements)

References 33 publications

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“…Comparing the new method to current OPA related studies (Pereira et al 2008; Kaspar et al 2009; Arrieta and Prats-Moya 2012) including analysis of complex biological matrices, it was possible to achieve a significant increase in sensitivity of chromatographic results. Overall sensitivity parameters of the method were also comparable to recent reports on sub -2 μm particle column (Mayer et al 2010) and core–shell (Song et al 2012a, b) amino acid analysis using different derivatization procedures and detection options. The obtained data show an overall improvement of detection sensitivity from μM to the nM range, which is a magnitude lower then LOD’s for OPA derivative analysis reported earlier (Wan et al 2000).…”

Section: Resultssupporting

confidence: 84%

“…Following on from that, we were aiming to overcome the limitation of sub -2 μm particles to special equipment caused by high column back pressure (Fekete et al 2012; Song et al 2012a, b) of about 700 bar in our case respectively. Therefore, we directly transferred chromatographic conditions to the application of a 2.6-μm core–shell C 18 column (Sunshell C18) with the same dimensions (150 mm × 2.1 mm).…”

Section: Resultsmentioning

confidence: 99%

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Simultaneous analysis of the non-canonical amino acids norleucine and norvaline in biopharmaceutical-related fermentation processes by a new ultra-high performance liquid chromatography approach

et al. 2013

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In this study, a precise and reliable ultra-high performance liquid chromatography (UHPLC) method for the simultaneous determination of non-canonical (norvaline and norleucine) and standard amino acids (aspartic acid, glutamic acid, serine, histidine, glycine, threonine, arginine, tyrosine, methionine, valine, phenylalanine, isoleucine, leucine) in biopharmaceutical-related fermentation processes was established. After pre-column derivatization with ortho-phthaldialdehyde and 2-mercaptoethanol, the derivatives were separated on a sub-2 μm particle C18 reverse-phase column. Identification and quantification of amino acids were carried out by fluorescence detection. To test method feasibility on standard HPLC instruments, the assay was properly transferred to a core–shell particle C18 reverse-phase column. The limits of detection showed excellent sensitivity by values from 0.06 to 0.17 pmol per injection and limits of quantification between 0.19 and 0.89 pmol. In the present study, the newly established UHPLC method was applied to a recombinant antibody Escherichia coli fermentation process for the analysis of total free amino acids. We were able to specifically detect and quantify the unfavorable amino acids in such complex samples. Since we observed trace amounts of norvaline and norleucine during all fermentation phases, an obligatory process monitoring should be considered to improve quality of recombinant protein drugs in future.

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Section: Resultssupporting

confidence: 84%

Section: Resultsmentioning

confidence: 99%

Simultaneous analysis of the non-canonical amino acids norleucine and norvaline in biopharmaceutical-related fermentation processes by a new ultra-high performance liquid chromatography approach

et al. 2013

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“…Although the retention ability of MonoTower column was similar to that of the MonoClad column, the NBD-amino acids could not be separated well. Hence, the separation conditions were optimized basically by changing the TFA concentration, which was effective on the separation of NBD-amino acids [13]. Finally, the optimized mobile phase and gradient conditions were as described in Experimental section.…”

Section: Separation Of Nbd-amino Acidsmentioning

confidence: 99%

Amino Acid Analysis Using a Cartridge-Type Monolithic Silica Column

2017

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In this study, the application of a new type of monolithic silica column to the analysis of amino acids in human plasma samples has been investigated. The monolithic silica column (3.0 × 150 mm, MonoTower C18) provided good separation of the amino acids derivatized with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F). Although the separation ability of this column was not much improved in comparison with the previous monolithic silica columns, it was similar with that of a 3 μm particle packed column. The column with length 150 mm had the same separation property as a tandem connection of columns of lengths 50 mm and 100 mm. After 1000 injections of human plasma samples, the peak widths for the NBD-amino acids were increased by only 8%. Thus, the newly developed monolithic silica column can be potentially applied for faster analysis and/or more efficient separation of biological compounds.

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“…The derivatization procedure of the aliphatic amines was described previously [25][26][27]. Briefly, 20 µL of the aliphatic amine solution and 40 µL of 10 mM the NBD-F solution were mixed with 180 µL of a 0.2 M borate buffer (pH 8.5).…”

Section: Derivatization Procedures Of Aliphatic Aminesmentioning

confidence: 99%

Retention and Bandwidth Predictions by Fast Gradient Elution Chromatography Using a Pillar Array Column

et al. 2016

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Previously, we have developed a gradient elution system for pillar array columns, which achieved faster separation than isocratic elution. In this study, we validated gradient elution in microchip liquid chromatography (LC) and investigated the retention and bandwidth predictions of fluorescently labeled aliphatic amines in fast gradient elution chromatography using semi-empirical retention models. The retention times and peak widths under different gradient elution programs were predicted by three solvent strength models and compared with the experimental results. The relative errors of prediction for the retention times and peak widths were below 14 % and 12 %, respectively. The results showed that the solvent strength models could be utilized for predicting the retention times and peak widths under gradient elution in the microchip LC system and that the gradient elution program for pillar array columns worked efficiently. The prediction by the retention model promises to be a potential tool for essential compound identification in biological samples.

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Rapid determination of amino acids in biological samples using a monolithic silica column

Cited by 29 publications

References 33 publications

Simultaneous analysis of the non-canonical amino acids norleucine and norvaline in biopharmaceutical-related fermentation processes by a new ultra-high performance liquid chromatography approach

Simultaneous analysis of the non-canonical amino acids norleucine and norvaline in biopharmaceutical-related fermentation processes by a new ultra-high performance liquid chromatography approach

Amino Acid Analysis Using a Cartridge-Type Monolithic Silica Column

Retention and Bandwidth Predictions by Fast Gradient Elution Chromatography Using a Pillar Array Column

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