Determination of Twenty Proteinogenic Amino Acids and Additives in Cultural Liquid by High-Performance Liquid Chromatography

Askretkov, A. D.; Klishin, A. A.; Зыбин, Д. И.; Орлова, Н. В.; Kholodova, A. V.; Lobanova, N. V.; Seregin, Yu. A.

doi:10.1134/s1061934820080031

Cited by 8 publications

(3 citation statements)

References 21 publications

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“…In conclusion, a pre‐column derivatization is a crucial tool for analyzing BCAAs due to its ability to enhance detection sensitivity and its versatility in accommodating various sample types and detectors. Table 1 shows a compilation of 19 commonly used derivatization reagents, along with a summary of their respective advantages and disadvantages [33–55]. The different reactions with AAs are described in Figure 2.…”

Section: Derivatization Approachesmentioning

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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Section: Derivatization Approachesmentioning

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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“…An Amino Acid 𝐻2𝑁 − 𝐶𝐻(𝑅) − 𝐶𝑂𝑂𝐻 However, this generalization does not encompass all amino acids; deviations include those with multiple carboxyl or amino groups, imino groups, alternative types of acid groups, and configurations where the amino group is not attached to the α-carbon (1,2). It is noteworthy that most amino acids contain at least one asymmetric carbon atom, enabling their existence in multiple optically active forms.…”

Section: Introductionmentioning

confidence: 99%

Spectrophotometric Study of Excitatory Amino Acids with Ortho-Phthalaldehyde

Baloch,

Ashraf,

Durani

et al. 2024

JHRR

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Background: Understanding the concentration of amino acids in biological samples is pivotal for numerous biochemical and clinical studies. Aspartic acid and glutamic acid, being excitatory amino acids, play crucial roles in the neurotransmission processes of the mammalian brain. Their precise quantification is essential for advancing our knowledge in neurochemistry and for the diagnosis of related disorders. Objective: This study aimed to develop a robust, accurate, and sensitive spectrophotometric method for quantifying aspartic and glutamic acids in solution, both in their native forms and when complexed with ortho-phthalaldehyde (OPA), a derivatization agent that enhances their absorbance properties. Methods: Utilizing a double-beam Hitachi 220 Spectrophotometer equipped with dual 1 cm silica cuvettes, we performed spectrophotometric analyses across a wavelength range of 165 to 1000 nm. Calibration curves for aspartic acid and glutamic acid, with and without OPA, were generated by plotting absorbance against concentration. Various solvents including methanol, ethanol, acetone, and acetonitrile were evaluated for their efficacy in dissolving the amino acids and their complexes. Linear regression analysis was employed to establish the relationship between concentration and absorbance, and the method's precision and accuracy were validated through replicate measurements at multiple concentrations. Results: The calibration curves exhibited excellent linearity for both amino acids, with and without OPA, across the tested concentration ranges. For aspartic acid, absorption maxima were observed at 201 nm (106,333.33 L mol^-1 cm^-1 molar absorptivity) and 229 nm (71,250 L mol^-1 cm^-1) for its OPA derivative. Glutamic acid showed absorption maxima at 198 nm (84,333.33 L mol^-1 cm^-1) and 216 nm (128,857.14 L mol^-1 cm^-1) for its OPA derivative. The correlation coefficients (R²) for the calibration curves ranged from 0.9954 to 0.9982, indicating a high degree of linearity and reliability of the method. Conclusion: The developed spectrophotometric method provides a reliable, precise, and sensitive approach for quantifying aspartic and glutamic acids. Its application is significant for biochemical analysis and has potential implications in clinical diagnostics, offering a promising tool for neurochemical studies.

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“…As amino acids are required in the growth of cells, amino acids are often included as a component in cell culture media. In terms of production, quality assurance and control of the exact composition of the cell culture media regarding the concentration of the various amino acids is an important application [ 21 , 22 , 23 ]. Unregulated control of concentration of amino acids in these cell culture media can lead to unstable metabolism and irreproducible results which may have dire consequences in clinical applications [ 24 ].…”

Section: Introductionmentioning

confidence: 99%

Split-Ring Resonator Based Sensor for the Detection of Amino Acids in Liquids

Dehning

Hitzemann

Gossmann

et al. 2023

Sensors

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Amino acids belong to the most important compounds for life. They are structural components of proteins and required for growth and maintenance of cells. Essential amino acids cannot be produced by the organism and must be ingested through the nutrition. Therefore, the detection of amino acids is of great interest when analyzing cell culture media and nutrition. In this work, we present a split-ring resonator as a simple but sensitive detector for amino acids. Split-ring resonators are RLC resonant circuits with a split capacitance and thus a resonance frequency that depends on the electromagnetic properties of a liquid sample at the split capacitance. Here, the split capacitance is an interdigital structure for highest sensitivity and covered with a fluidic channel for flow through experiments. First measurements with a vector network analyzer show detection limits in the range from 105 µM for glutamic acid to 1564 µM for isoleucine, depending on the electromagnetic properties of the tested amino acids. With an envelope detector for continuous recording of the resonance frequency, the split-ring resonator can be used in ion chromatography. At a flow rate of 0.5 mL/min, it reaches limits of detection of 485 µM for aspartic acid and 956 µM for lysine.

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Determination of Twenty Proteinogenic Amino Acids and Additives in Cultural Liquid by High-Performance Liquid Chromatography

Cited by 8 publications

References 21 publications

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

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

Spectrophotometric Study of Excitatory Amino Acids with Ortho-Phthalaldehyde

Split-Ring Resonator Based Sensor for the Detection of Amino Acids in Liquids

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