Determination of underivatized amino acids in microsamples of a yeast nutritional supplement by LC-MS following microwave assisted acid hydrolysis

Aviram, Lilach Yishai; McCooeye, Margaret; Mester, Zoltán

doi:10.1039/c6ay00407e

Cited by 9 publications

(3 citation statements)

References 44 publications

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“…The LODs and LOQs for the 20 compounds were in the range of 0.0036–10.00 and 0.016–20.00 ng/mL, respectively. Such levels of sensitivity for amino acid detection are noticeably higher than those of other reported techniques using MRM methods . The maximum sensitivity of MRM detection with the Waters Xevo TQ‐S quadrupole mass spectrometer and sharp peak shapes were observed using 85% v/v acetonitrile with 1.7 mmol/L ammonium acetate and 0.1% formic acid as the solvent.…”

Section: Resultsmentioning

confidence: 88%

“…Ideally, such a method would reduce data variability due to derivative instability, side reactions, and reagent interferences as well as be fast, reliable, and simple. As MS methods have improved, MS/MS‐based approaches have facilitated the development of single‐run LC‐MS‐based approaches for identifying and quantifying a range of amino acids and their metabolic products in a variety of media . The development of these approaches has dramatically reduced processing times and reduced the number of synthetic steps .…”

Section: Introductionmentioning

confidence: 99%

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A rapid, sensitive, and widely applicable method for quantitative analysis of underivatized amino acids in different biological matrices by UHPLC‐MS/MS

Guo

Yang

et al. 2019

J of Separation Science

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A rapid, sensitive, and widely applicable method for the simultaneous quantitative analysis of 20 underivatized amino acids in different biological matrices, including serum, plasma, and tissue homogenates, using ultra high performance liquid chromatography with tandem mass spectrometry was developed and validated.Only 4 μL of serum, plasma, or tissue homogenate was extracted with 996 μL of solution (1.7 mM ammonium formate in 85% acetonitrile containing 0.1% formic acid) containing 100 ng/mL phenylalanine-d5 as an internal standard without any further derivatization step. In addition, the matrix effects were small because a large volume of extraction solution was used. The total run time including reequilibration was 13 min. The results of linearity, accuracy, repeatability, precision, limits of detection, limits of quantification, and sample stability were sufficient to allow the measurement of the amino acids in different biological matrices. We conclude that our method is rapid, sensitive, and widely applicable and represents an improvement over other currently available technologies. K E Y W O R D Samino acids, biological matrices, tandem mass spectrometry, ultra high performance liquid chromatography Article Related Abbreviations: CE, collision energy; CV, cone voltage; IS, internal standard; MRM, multiple reaction monitoring. selective methods for the identification and quantification of amino acids is important.Amino acids have a variety of structures and physicochemical properties; therefore, quantitative methods must be robust, sensitive, and amenable to a number of different media. Recently, several technologies have been developed for the analysis of amino acids, including GC-MS [7,8], , stable isotope labelling [14][15][16], and absorbance methodologies [17]. Because of the complexity of media such as urine, plasma, and cerebrospinal fluid as well as the diversity of amino acid properties, methods for quantifying amino acids are generally complex and require multiple clean-up steps to improve the sensitivity and selectivity. For

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Section: Resultsmentioning

confidence: 88%

Section: Introductionmentioning

confidence: 99%

A rapid, sensitive, and widely applicable method for quantitative analysis of underivatized amino acids in different biological matrices by UHPLC‐MS/MS

Guo

Yang

et al. 2019

J of Separation Science

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“…Liquid chromatography does not require derivatizations to enhance volatility and when coupled with tandem mass spectrometry (LC–MS/MS) provides a burgeoning set of hyphenated techniques to complement polar compound analysis. LC–MS/MS has high selectivity and is sensitive to detection of both derivatized [21] and underivatized amino acids [22, 23]; however, the coupling of LC with MS is restricted to compatible volatile solvents unless additional desalting processes are introduced. LC–MS/MS has enabled rapid throughput with reverse phase columns to resolve most amino acids [24].…”

Section: Introductionmentioning

confidence: 99%

Accurate and efficient amino acid analysis for protein quantification using hydrophilic interaction chromatography coupled tandem mass spectrometry

Kambhampati

Evans

et al. 2019

Plant Methods

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Background Methods used to quantify protein from biological samples are often inaccurate with significant variability that requires care to minimize. The errors result from losses during protein preparation and purification and false detection of interfering compounds or elements. Amino acid analysis (AAA) involves a series of chromatographic techniques that can be used to measure protein levels, avoiding some difficulties and providing specific compositional information. However, unstable derivatives, that are toxic and can be costly, incomplete reactions, inadequate chromatographic separations, and the lack of a single hydrolysis method with sufficient recovery of all amino acids hinder precise protein quantitation using AAA. Results In this study, a hydrophilic interaction chromatography based method was used to separate all proteinogenic amino acids, including isobaric compounds leucine and isoleucine, prior to detection by multiple reaction monitoring with LC–MS/MS. Through inclusion of commercially available isotopically labeled ( 13 C, 15 N) amino acids as internal standards we adapted an isotopic dilution strategy for amino acid-based quantification of proteins. Three hydrolysis methods were tested with ubiquitin, bovine serum albumin, (BSA), and a soy protein biological reference material (SRM 3234; NIST) resulting in protein estimates that were 86–103%, 82–94%, and 90–99% accurate for the three protein samples respectively. The methane sulfonic acid hydrolysis approach provided the best recovery of labile amino acids including: cysteine, methionine and tryptophan that are challenging to accurately quantify. Conclusions Accurate determination of protein quantity and amino acid composition in heterogeneous biological samples is non-trivial. Recent advances in chromatographic phases and LC–MS/MS based methods, along with the availability of isotopic standards can minimize difficulties in analysis and improve protein quantitation. A robust method is described for high-throughput protein quantification and amino acid compositional analysis. Since accurate measurement of protein quality and quantity are a requirement for many biological studies that relate to crop improvement or more generally, our understanding of metabolism in living systems, we envision this method will have broad applicability. Electronic supplementary material The online version of this article (10.1186/s13007-019-0430-z) contains supplementary material, which is available to authorized users.

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A new green approach to l-histidine and β-alanine analysis in dietary supplements using rapid and simple contactless conductivity detection integrated with high-resolution glass-microchip electrophoresis

Pukleš,

Páger,

Sakač

et al. 2024

Anal Bioanal Chem

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Determination of underivatized amino acids in microsamples of a yeast nutritional supplement by LC-MS following microwave assisted acid hydrolysis

Abstract: In this paper we describe a rapid method for microscale microwave assisted acid hydrolysis followed by quantitative amino acid analysis, using liquid chromatography mass spectrometry UPLC-ESI-MS (QTOF) without derivatization.

Cited by 9 publications

References 44 publications

A rapid, sensitive, and widely applicable method for quantitative analysis of underivatized amino acids in different biological matrices by UHPLC‐MS/MS

A rapid, sensitive, and widely applicable method for quantitative analysis of underivatized amino acids in different biological matrices by UHPLC‐MS/MS

Accurate and efficient amino acid analysis for protein quantification using hydrophilic interaction chromatography coupled tandem mass spectrometry

A new green approach to l-histidine and β-alanine analysis in dietary supplements using rapid and simple contactless conductivity detection integrated with high-resolution glass-microchip electrophoresis

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