Simultaneous determination of aromatic amino acids in human blood plasma by capillary electrophoresis with UV‐absorption detection

Forteschi, Mauro; Sotgia, Salvatore; Assaretti, Stefano; Arru, Dionigia; Cambedda, Debora; Sotgiu, Elisabetta; Zinellu, Angelo; Carru, Ciriaco

doi:10.1002/jssc.201500038

Cited by 23 publications

(12 citation statements)

References 37 publications

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“…In blood, there is approximately 60-80 μM each AAA, totaling 200 μM (Forteschi et al, 2015). In the kidneys, there are comparable amounts (Li, Tang, & Mu, 2011).…”

Section: Figurementioning

confidence: 99%

A new host cell internalisation pathway for SadA‐expressing staphylococci triggered by excreted neurochemicals

Luqman

Ebner

Reichert

et al. 2019

Cellular Microbiology

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Staphylococcus aureus is a facultative intracellular pathogen that invades a wide range of professional and nonprofessional phagocytes by triggering internalisation by interaction of surface‐bound adhesins with corresponding host cell receptors. Here, we identified a new concept of host cell internalisation in animal‐pathogenic staphylococcal species. This new mechanism exemplified by Staphylococcus pseudintermedius ED99 is not based on surface‐bound adhesins but is due to excreted small neurochemical compounds, such as trace amines (TAs), dopamine (DOP), and serotonin (SER), that render host cells competent for bacterial internalisation. The neurochemicals are produced by only one enzyme, the staphylococcal aromatic amino acid decarboxylase (SadA). Here, we unravelled the mechanism of how neurochemicals trigger internalisation into the human colon cell line HT‐29. We found that TAs and DOP are agonists of the α2‐adrenergic receptor, which, when activated, induces a cascade of reactions involving a decrease in the cytoplasmic cAMP level and an increase in F‐actin formation. The signalling cascade of SER follows a different pathway. SER interacts with 5HT receptors that trigger F‐actin formation without decreasing the cytoplasmic cAMP level. The neurochemical‐induced internalisation in host cells is independent of the fibronectin‐binding protein pathway and has an additive effect. In a sadA deletion mutant, ED99ΔsadA, internalisation was decreased approximately threefold compared with that of the parent strain, and treating S. aureus USA300 with TAs increased internalisation by approximately threefold.

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“…In blood, there is approximately 60-80 μM each AAA, totaling 200 μM (Forteschi et al, 2015). In the kidneys, there are comparable amounts (Li, Tang, & Mu, 2011).…”

Section: Figurementioning

confidence: 99%

A new host cell internalisation pathway for SadA‐expressing staphylococci triggered by excreted neurochemicals

Luqman

Ebner

Reichert

et al. 2019

Cellular Microbiology

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“…The α-CD did not improve resolution whereas β-CD improved Ala isomers resolution. For this, β-CD concentration (2.5, 5, 7.5, 10, and 15 mM) as well as the borate buffer concentration (10,20,30,50,75, and 100 mM) were optimized. Best results were obtained using the following BGE: 100 mM sodium tetraborate and 10 mM β-cyclodextrin at pH 9.4.…”

Section: Analysis Of Labeled Amino Acids By Ce With Lif Detectionmentioning

confidence: 99%

“…Several analytical methods for analyzing amino acids have been reported in the literature based on chromatographic [5][6][7][8][9][10][11] and electrophoretic methodologies [12][13][14][15][16][17][18]. Different detectors can be used such as UV (UV/VIS) [5,19,20], fluorescence [15,[21][22][23][24][25][26], conductivity [18], and MS detection [11,16,[27][28][29][30]. Recently, CE-MS was used for the identification of amino acids due to its capability to differentiate overlapping peaks with distinct mass-to-charge ratios [29,30].…”

mentioning

confidence: 99%

Microalgae amino acid extraction and analysis at nanomolar level using electroporation and capillary electrophoresis with laser‐induced fluorescence detection

Nehmé

Atieh

Fayad

et al. 2016

J of Separation Science

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Amino acids play a key role in food analysis, clinical diagnostics, and biochemical research. Capillary electrophoresis with laser-induced fluorescence detection was used for the analysis of several amino acids. Amino acid labeling with fluorescein isothiocyanate was conducted using microwave-assisted derivatization at 80°C (680 W) during only 150 s. Good electrophoretic resolution was obtained using a background electrolyte composed of sodium tetraborate buffer (100 mM; pH 9.4) and β-cyclodextrin (10 mM), and the limits of quantification were 3-30 nM. The developed capillary electrophoresis with laser-induced fluorescence method was used to analyze amino acids in Dunaliella salina green algae grown under different conditions. A simple extraction technique based on electroporation of the cell membrane was introduced. A home-made apparatus allowed the application of direct and alternating voltages across the electrochemical compartment containing a suspension of microalgae in distilled water at 2.5 g/L. A direct voltage of 12 V applied for 4 min gave the optimum extraction yield. Results were comparable to those obtained with accelerated-solvent extraction. The efficiency of electroporation in destroying microalgae membranes was shown by examining the algae surface morphology using scanning electron microscopy. Stress conditions were found to induce the production of amino acids in Dunaliella salina cells.

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“…Their simultaneous quantification is possible with CE. Since no derivatization is required, the UV detection is used .…”

Section: Ce In Clinical Biochemistrymentioning

confidence: 99%

Recent advances in amino acid analysis by capillary electromigration methods, 2013–2015

et al. 2015

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We describe the most important research articles published on amino acid analysis using CE during the period from June 2013 to May 2015, and follows the format of the previous articles published in electrophoresis the new developments in amino acid analysis with CE are mainly describing improvements in detection means and injection methods. Enantiomeric separation developments are still important. Focusing the applications, we describe the neurochemical and clinical works, but also the metabolomic studies for which the publication number increase greatly. Finally, works focused on amino acids in food and agricultural applications are described.

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Simultaneous determination of aromatic amino acids in human blood plasma by capillary electrophoresis with UV‐absorption detection

Cited by 23 publications

References 37 publications

A new host cell internalisation pathway for SadA‐expressing staphylococci triggered by excreted neurochemicals

A new host cell internalisation pathway for SadA‐expressing staphylococci triggered by excreted neurochemicals

Microalgae amino acid extraction and analysis at nanomolar level using electroporation and capillary electrophoresis with laser‐induced fluorescence detection

Recent advances in amino acid analysis by capillary electromigration methods, 2013–2015

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