Quantitation of non-derivatized free amino acids for detecting inborn errors of metabolism by incorporating mixed-mode chromatography with tandem mass spectrometry

DeArmond, Patrick D.; Bunch, Dustin R.

doi:10.1016/j.jmsacl.2022.05.002

Cited by 9 publications

(11 citation statements)

References 24 publications

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“…Our dual-column U(H)PLC-MRM-MS method provided lower LLOD and LLOQ concentrations for all amino acids analyzed and was 15 times faster than the gold standard (run time: 119 min) with total a run time of 7.9 min. Choi et al 32 , Liu et al 47 and DeArmond et al 33 recently developed HPLC-MRM- MS methods based on mixed-mode chromatography using hydrophilic interaction chromatography (HILIC) and cation exchange (CEX) properties to separate and quantify free amino acids. They achieved LLOQ values in the low µmolar range with analysis times ranging from 13 min 47 to 15 min 33 and 22 min 32 , respectively.…”

Section: Resultsmentioning

confidence: 99%

Automated liquid handling extraction and rapid quantification of underivatized amino acids and tryptophan metabolites from human serum and plasma using dual-column U(H)PLC-MRM-MS and its application to prostate cancer study

Kipura,

Hotze,

Hofer

et al. 2024

Preprint

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Free amino acids (AAs) and their metabolites are important building blocks, energy sources and signaling molecules associated with various pathological phenotypes. The quantification of AA and tryptophan (TRP) metabolites in human serum and plasma is therefore of great diagnostic interest. Robust and reproducible sample extraction and processing workflows as well as rapid, sensitive absolute quantification of AA and TRP metabolites are required to identify candidate biomarkers and to improve current screening methods. We developed a validated semi-automated extraction and sample processing workflow using a robotic liquid handling platform and a rapid method for the absolute quantification of 20 free, underivatized AAs and 6 TRP metabolites using dual-column U(H)PLC-MRM-MS. The automated extraction and sample preparation workflow is designed for use in a 96-well plate format, allowing robust and reproducible high sample throughput without the need for further SPE, evaporation and/or buffer exchange. Samples extracted from serum and/or plasma in 96-well plates can be transferred directly to the U(H)PLC autosampler. The dual-column U(H)PLC-MRM-MS method, using a mixed-mode reversed-phase anion exchange column with formic acid as mobile phase modifier and a high-strength silica reversed-phase column with difluoroacetic acid as mobile phase additive, provided absolute quantification with nanomolar lower limits of quantification (LLOQ) for all metabolites except glycine (LLOQ: 2.46 micromolar) in only 7.9 minutes. The semi-automated extraction workflow and dual-column U(H)PLC-MRM-MS method was applied to a human prostate cancer study and was shown to discriminate between treatment regimens and to identify amino acids responsible for the statistical separation between healthy controls and prostate cancer patients on active surveillance.

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Section: Resultsmentioning

confidence: 99%

Automated liquid handling extraction and rapid quantification of underivatized amino acids and tryptophan metabolites from human serum and plasma using dual-column U(H)PLC-MRM-MS and its application to prostate cancer study

Kipura,

Hotze,

Hofer

et al. 2024

Preprint

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“…The labeled AA are then separated using RPLC, IEX, or HILIC methods. The direct AA analysis is a simpler alternative to methods based on derivatization (DeArmond & Bunch, 2022). A simple and fast method combining pre‐column derivatization with RPLC‐MS analysis for AA quantification in protein and biopharmaceutical samples has been proposed.…”

Section: Ms‐based Methods In the Analysis Of Biopharmaceuticalsmentioning

confidence: 99%

What is the role of current mass spectrometry in pharmaceutical analysis?

Khalikova

Jireš

Horáček

et al. 2023

Mass Spectrometry Reviews

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The role of mass spectrometry (MS) has become more important in most application domains in recent years. Pharmaceutical analysis is specific due to its stringent regulation procedures, the need for good laboratory/manufacturing practices, and a large number of routine quality control analyses to be carried out. The role of MS is, therefore, very different throughout the whole drug development cycle. While it dominates within the drug discovery and development phase, in routine quality control, the role of MS is minor and indispensable only for selected applications. Moreover, its role is very different in the case of analysis of small molecule pharmaceuticals and biopharmaceuticals. Our review explains the role of current MS in the analysis of both small‐molecule chemical drugs and biopharmaceuticals. Important features of MS‐based technologies being implemented, method requirements, and related challenges are discussed. The differences in analytical procedures for small molecule pharmaceuticals and biopharmaceuticals are pointed out. While a single method or a small set of methods is usually sufficient for quality control in the case of small molecule pharmaceuticals and MS is often not indispensable, a large panel of methods including extensive use of MS must be used for quality control of biopharmaceuticals. Finally, expected development and future trends are outlined.

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“…The Intrada AAs column was specially designed for AAs analysis, allowing the separation of Leu and Ile without the need for derivatization. DeArmond et al [148] F I G U R E 5 1. Leucine isomers in plasma.…”

Section: Mixed-mode Chromatographymentioning

confidence: 99%

“…It is a simple analytical method that requires a short analysis time, and the separation of the isomers is significantly better than previously reported [145,146] (see Figure 5). DeArmond's study [148] showed that ACN and water are more conducive to the separation and ionization of Leu and Ile than MeOH and that the retention time is reduced. The results were compared with an AAs analyzer (Hitachi L-8800).…”

Section: Mixed-mode Chromatographymentioning

confidence: 99%

“…DeArmond et al. [148] used Imtakt Intrada columns (containing hydrophilic and cation‐exchange ligands embedded within the stationary phase) to efficiently separate 39 AAs from human plasma and urine within 15 min, where the isobaric compounds (i.e., Allo‐Ile, Leu, and Ile) were separated (R s more than 1). Similar to what was shown in Figure 1, in MRM transition, the same qualitative ions (132 > 86) were used, but different quantitative ions were used for separation (132 > 69 for Allo‐Ile and Ile, 132 > 43 for Leu).…”

Section: Approaches Without Derivatizationmentioning

confidence: 99%

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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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Quantitation of non-derivatized free amino acids for detecting inborn errors of metabolism by incorporating mixed-mode chromatography with tandem mass spectrometry

Cited by 9 publications

References 24 publications

Automated liquid handling extraction and rapid quantification of underivatized amino acids and tryptophan metabolites from human serum and plasma using dual-column U(H)PLC-MRM-MS and its application to prostate cancer study

Automated liquid handling extraction and rapid quantification of underivatized amino acids and tryptophan metabolites from human serum and plasma using dual-column U(H)PLC-MRM-MS and its application to prostate cancer study

What is the role of current mass spectrometry in pharmaceutical analysis?

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

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