Rapid and reliable analysis of underivatized amino acids in urine using tandem mass spectrometry

Fernández-del-Campo-García, María Teresa; Casas-Ferreira, Ana María; Rodríguez‐Gonzalo, Encarnación; Cordero, Bernardo Moreno; Pavón, José Luis Pérez

doi:10.1016/j.microc.2021.106914

Cited by 7 publications

(3 citation statements)

References 27 publications

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“…Several publications have reported AA analysis in foods and biological fluids using GC–MS, capillary electrophoresis, HILIC-MS/MS, ion-paired LC–MS/MS, electrochemical detection (CE), ion exchange chromatography, and HPLC. , Most of these methods depend on pre- or postcolumn derivatization to improve the detection sensitivity. However, elimination of derivatization steps attracts many researchers, and many studies have described the analysis of AAs without derivatization steps such as underivatized determination of free AAs in honey using LC–MS/MS, branched-chain AAs in maple syrup urine disease using LC–MS/MS, evaluation embryo viability using CE-MS, urine analysis by LC–MS/MS, ion exchange, CE-MS coupled to a porous layer-gold nanoparticle-modified chiral column, AAs and peptides in nutritive mixtures in the total parenteral nutrition solution, and online coupling of UV, fluorescence, and electrochemical detection . The elimination of the derivatization step is advantageous for many reasons as the selection of an appropriate derivatization reagent is challenging as the reagents differ in their selectivity toward each amino acid; also, additional peaks usually appear in chromatograms, side reactions usually occur, complete derivatization cannot be ensured, long sample preparation time is required, and unstable derivatizations occur.…”

Section: Resultsmentioning

confidence: 99%

Underivatized Amino Acid Chromatographic Separation: Optimized Conditions for HPLC-UV Simultaneous Quantification of Isoleucine, Leucine, Lysine, Threonine, Histidine, Valine, Methionine, Phenylalanine, Tryptophan, and Tyrosine in Dietary Supplements

2022

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Amino acids (AAs) are considered as the building blocks of life. Unlike nonessential AAs, the human body cannot synthesize essential AAs and should be supplied in food or dietary supplements. The aim of the work is simultaneous HPLC-UV determination of 10 structurally related AAs without pre- or postderivatization in powdered dietary supplements (PDSs). This was challenging, especially because PDS has no standardized procedures for its quality control. HPLC-UV chromatograms of the 10 AAs were recorded using a gradient elution of the mobile phase on a CLC-C18 column at 225 nm. The elution started with 100% of phosphate buffer (pH 7.4, 10 mM) for 10 min; then, the concentration of acetonitrile increased linearly to reach 50% for another 15 min at room temperature. Good separation was achieved within a 25 min run time without pre- or postderivatization. The method was carefully validated according to the ICH guidelines over the linearity range of 100–200, 50–200, 20–150, 50–400, 20–250, 75–175, 50–250, 50–250, 50–300, and 5–100 μg/mL for l -lysine, l -threonine, l -histidine, l -valine, l -methionine, l -isoleucine, l -leucine, l -tyrosine, l -phenylalanine, and l -tryptophan, respectively, with mean recoveries ranges between 98.91 and 100.77. The method was found to be precise, and the relative standard deviation (RSD) was found to be between 0.28 and 1.92 with recoveries between 97.91 and 101.11. The method was found to be robust that resists deliberate changes in pH, flow rate, and mobile-phase percentages. It was successfully applied for the analysis of PDSs. The proposed method could be very useful for the quality control of the 10 structurally related AAs during their synthesis and for testing raw materials and pharmaceutical preparations.

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Section: Resultsmentioning

confidence: 99%

Underivatized Amino Acid Chromatographic Separation: Optimized Conditions for HPLC-UV Simultaneous Quantification of Isoleucine, Leucine, Lysine, Threonine, Histidine, Valine, Methionine, Phenylalanine, Tryptophan, and Tyrosine in Dietary Supplements

2022

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“…As part of their study, the authors used LC-MS and GC-MS to determine the concentration of AAs in human urine. Compared with LC-MS, GC-MS has obvious advantages regarding analysis time; the whole process takes 2.8 min (1.5 min for analysis) [128].…”

Section: Ion-pair Chromatographymentioning

confidence: 99%

Overview of chromatographic and electrophoretic methods for the determination of branched‐chain amino acids

Yin

Adams

Schepdael

2023

J of Separation Science

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The significance of branched‐chain amino acids in diseases was clearly shown over the years. This review aims to describe the available techniques for their analytical determination. The article provides examples of the use of various analytical methods. The methods are divided into two categories: derivatization and non‐derivatization approaches. Separation is achieved through different chromatography or capillary electrophoresis techniques and can be combined with different detectors such as flame ionization, ultraviolet, fluorescence, and mass spectrometry. It compares the application of various derivatization reagents or detection as such for different detectors.

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“…The run duration was 35 min, the flow rate was 150 μL/min, the injection volume was 5 μL, and the mobile phase was a combination of UHQ water (solution A) and (solution B) Methanol with 0.1% Heptafluorobutyric anhydride (v/v), respectively. The detected AAs were asparagine, serine, aspartic acid, glycine, threonine, alanine, proline, His, lysine, valine, arginine, methionine, leucine, Tyr, phenylalanine, and Trp, and their RT was in the range of 5.3–15 min [29]. Using an LC/MS technique, Fleszar et al.…”

Section: Bioanalytical Approachesmentioning

confidence: 99%

A comprehensive review on recent trends in amino acids detection through analytical techniques

Kaur,

Rangra,

Chawla

2023

Separation Science Plus

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The process of growth, development, and reproduction cannot take place in a live body without the presence of amino acids (AAs) since they are necessary for appropriate functioning. Detection of AAs in pharmaceutical and food samples is required to ensure quality, quantity, and efficacy. The AAs generally do not contain chromophores; hence their chromatographic analysis requires derivatization. The research papers published in the last few years were used to compile this manuscript. This manuscript covers various analytical, bioanalytical, and electrochemical approaches used in the identification of AAs. The analytical techniques like high‐performance liquid chromatography, ultra‐performance liquid chromatography, etc., and the hyphenated analytical techniques like liquid chromatography‐mass spectrometry, ultra‐performance liquid chromatography‐electrospray tandem mass spectrometry, and hydrophilic interaction ultra‐performance liquid chromatography are also discussed. This manuscript also briefly outlines the electrochemical analytical techniques including sensors and biosensors. This review article will be useful to the researcher working in the area of the development of analytical techniques for the detection of AAs.

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Rapid and reliable analysis of underivatized amino acids in urine using tandem mass spectrometry

Cited by 7 publications

References 27 publications

Underivatized Amino Acid Chromatographic Separation: Optimized Conditions for HPLC-UV Simultaneous Quantification of Isoleucine, Leucine, Lysine, Threonine, Histidine, Valine, Methionine, Phenylalanine, Tryptophan, and Tyrosine in Dietary Supplements

Underivatized Amino Acid Chromatographic Separation: Optimized Conditions for HPLC-UV Simultaneous Quantification of Isoleucine, Leucine, Lysine, Threonine, Histidine, Valine, Methionine, Phenylalanine, Tryptophan, and Tyrosine in Dietary Supplements

Overview of chromatographic and electrophoretic methods for the determination of branched‐chain amino acids

A comprehensive review on recent trends in amino acids detection through analytical techniques

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