Quantification of tryptophan in plasma by high performance liquid chromatography

Pinhati, Renata Romanholi; Polonini, Hudson; Brandão, Marcos Antônio Fernandes; Raposo, Nádia Rezende Barbosa; Corchs, Felipe; Gattaz, Wagner F.

doi:10.1590/s0100-40422012000300032

Cited by 8 publications

(16 citation statements)

References 10 publications

(14 reference statements)

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“…Proteins in plasma are frequently removed through a pretreatment of sample with acids and by SPE. Deproteinization with perchloric acid (PCA) and trichloroacetic acid (TCA) might be used for this purpose. The sample mixed with acid is shaken and centrifuged to separate precipitated proteins from Trp and kynurenines present in supernatant.…”

Section: Sample Preparation For Chromatographic Analysis Of Kynureninesmentioning

confidence: 99%

“…However, indole derivatives are sensitive to acidic conditions and precipitation with TCA lowers the Kyn signal . Some authors had also proposed the use of other acids like sulfosalicylic acid , hydrochloric acid , mixture of ascorbic acid, and PCA or TCA with addition of hydrochloric acid or acetonitrile . The above agents seem not to be optimal for kynurenines quantification and better choice might be deproteinization with methanol , ethanol (chilled or room temperature), and mixture of ammonium acetate in methanol or ammonium acetate in water .…”

Section: Sample Preparation For Chromatographic Analysis Of Kynureninesmentioning

confidence: 99%

“…It is based on the nonpolar, polar, ion exchange (cation and anion), and mixed mode interactions of sorbent with an analyte dissolved in liquid phase and subsequent elution with an appropriate solvent. Dowex‐50W cation exchange , SepPak , and other cartridges have been used for extraction of kynurenines. Implementing of the automated on‐line SPE (using propylsulfonic cartridges) followed by LC–MS/MS shortens the time of analysis reported for quantification of ʟ‐Trp, ʟ‐Kyn, and 3HKyn in human plasma .…”

Section: Sample Preparation For Chromatographic Analysis Of Kynureninesmentioning

confidence: 99%

“…LC has been often used for this purpose. There are methods employing various detection modalities: UV absorbance , fluorescence , electrochemical methods as well as MS . The LC separation is mostly achieved on octadecyl silica (ODS) columns.…”

Section: Chromatographic Analysis Of Tryptophan Metabolitesmentioning

confidence: 99%

“…The LC separation is mostly achieved on octadecyl silica (ODS) columns. The mobile phases usually contain a small addition of acetonitrile (see Table ) that shortens a retention time and enhances Trp signal . HPLC–UV is an attractive method for clinical applications.…”

Section: Chromatographic Analysis Of Tryptophan Metabolitesmentioning

confidence: 99%

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Chromatographic analysis of tryptophan metabolites

Sadok

Gamian

Staniszewska

2017

J of Separation Science

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The kynurenine pathway generates multiple tryptophan metabolites called collectively kynurenines and leads to formation of the enzyme cofactor nicotinamide adenine dinucleotide. The first step in this pathway is tryptophan degradation, initiated by the rate‐limiting enzymes indoleamine 2,3‐dioxygenase, or tryptophan 2,3‐dioxygenase, depending on the tissue. The balanced kynurenine metabolism, which has been a subject of multiple studies in last decades, plays an important role in several physiological and pathological conditions such as infections, autoimmunity, neurological disorders, cancer, cataracts, as well as pregnancy. Understanding the regulation of tryptophan depletion provide novel diagnostic and treatment opportunities, however it requires reliable methods for quantification of kynurenines in biological samples with complex composition (body fluids, tissues, or cells). Trace concentrations, interference of sample components, and instability of some tryptophan metabolites need to be addressed using analytical methods. The novel separation approaches and optimized extraction protocols help to overcome difficulties in analyzing kynurenines within the complex tissue material. Recent developments in chromatography coupled with mass spectrometry provide new opportunity for quantification of tryptophan and its degradation products in various biological samples. In this review, we present current accomplishments in the chromatographic methodologies proposed for detection of tryptophan metabolites and provide a guide for choosing the optimal approach.

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Section: Sample Preparation For Chromatographic Analysis Of Kynureninesmentioning

confidence: 99%

Section: Sample Preparation For Chromatographic Analysis Of Kynureninesmentioning

confidence: 99%

Section: Sample Preparation For Chromatographic Analysis Of Kynureninesmentioning

confidence: 99%

Section: Chromatographic Analysis Of Tryptophan Metabolitesmentioning

confidence: 99%

Section: Chromatographic Analysis Of Tryptophan Metabolitesmentioning

confidence: 99%

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Chromatographic analysis of tryptophan metabolites

Sadok

Gamian

Staniszewska

2017

J of Separation Science

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Identification of biomarkers on kidney failure by Raman spectroscopy

Gulyamov

Shamshiddinova

Bae

et al. 2021

J Raman Spectroscopy

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Chronic kidney disease (CKD) is a global health problem. Our study employed Raman spectroscopy for the first time to analyze potential biomarker such as tryptophan (Trp) on kidney failure and show the difference between normal and CKD groups. Adenine model of CKD characterized most of the physiological changes observed in human CKD. For that, 6‐week‐old Wistar male rats were assigned to the two groups: the control group, 0.5% carboxymethyl cellulose (CMC) was administered; the CKD group, adenine was administered to rats at a dose 600 mg/kg for 10 days. The Raman spectra were obtained in the spectral range of 100–2500 cm−1. The excitation source was 532 nm, the laser power was ~40 mW, and the analyzed tissue area was 10.02 × 82.12 μm2. The results showed a significant reduction in the case of Raman peak intensities at 748, 1359, 1554, and 1636 cm−1, which can be assigned as Trp. Moreover, it was shown that tyrosine at 1169 cm−1 and phenylalanine at 1001 cm−1 decreased in the CKD group. Trp and kynurenine (Kyn) levels in kidney tissue and plasma were analyzed by using the high‐performance liquid chromatography (HPLC). The Trp concentrations in plasma and kidney tissue significantly decreased in CKD 7.4 μg/ml and 0.1 μg/mg, respectively. However, Kyn level was not a statistically significant change in kidney tissue. In this work, we showed that Raman spectroscopy usage in CKD model and Raman spectroscopy can be considered a promising technique for semi‐quantitative analysis of potential biomarkers such as phenylalanine, tyrosine, and Trp of renal function because of the little or no need for sample preparation, and it is easily observing several biomarkers at the same time. The utilization of multivariate analysis method, such as partial least square analysis can be considered as a promising tool for the discrimination of control and CKD groups.

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Quantification of IDO1 enzyme activity in normal and malignant tissues

Zhai

Ladomersky

Bell

et al. 2019

Methods in Enzymology

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Indoleamine 2,3-dioxygenase 1 (IDO1) catalyzes the first and rate-limiting reaction of Ltryptophan (Trp) conversion into L-kynurenine (Kyn). The depletion of Trp, and the accumulation of Kyn have been proposed as mechanisms that contribute to the suppression of the immune response-primarily evidenced by in vitro study. IDO1 is therefore considered to be an immunosuppressive modulator and quantification of IDO1 metabolism may be critical to understanding its role in select immunopathologies, including autoimmune-and oncologicalconditions, as well as for determining the potency of IDO1 enzyme inhibitors. Because tryptophan 2,3-dioxygenase (TDO), and to a significantly lesser extent, IDO2, also catabolize Trp into Kyn, it's important to differentiate the contribution of each enzyme to Trp catabolism and Kyn generation. Moreover, a great variety of detection methods have been developed for the quantification of Trp metabolites, but choosing the suitable protocol remains challenging. Here, we review the differential expression of IDO1/TDO/IDO2 in normal and malignant tissues, followed by a comprehensive analysis of methodologies for quantifying Trp and Kyn in vitro and in vivo, with an emphasis on the advantages/disadvantages for each application.

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Quantification of tryptophan in plasma by high performance liquid chromatography

Cited by 8 publications

References 10 publications

Chromatographic analysis of tryptophan metabolites

Chromatographic analysis of tryptophan metabolites

Identification of biomarkers on kidney failure by Raman spectroscopy

Quantification of IDO1 enzyme activity in normal and malignant tissues

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