Separation and detection of D-/L-serine by conventional HPLC

Shikanai, Hiroki; Ikimura, Kazuko; Miura, Mitsutoshi; Shindo, Tsugumi; Watarai, Akane; Izumi, Tohru

doi:10.1016/j.mex.2022.101752

Cited by 2 publications

(2 citation statements)

References 13 publications

(13 reference statements)

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“…Adherence to ethical guidelines was paramount throughout the study, with protocols designed to align with the principles outlined in the Helsinki Declaration, despite the absence of direct clinical or human subject involvement. Data collection was robust and systematic, with spectrophotometric readings statistically analyzed using SPSS version 25, enabling a detailed evaluation of the interaction between amino acids and the OPA reagent (13). This study's framework, characterized by stringent experimental controls and a commitment to analytical accuracy, establishes a vital bridge between basic biochemistry and potential clinical diagnostics.…”

Section: Methodsmentioning

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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Section: Methodsmentioning

confidence: 99%

Spectrophotometric Study of Excitatory Amino Acids with Ortho-Phthalaldehyde

Baloch,

Ashraf,

Durani

et al. 2024

JHRR

View full text Add to dashboard Cite

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“…For the detection of l ‐Ser, the reaction supernatant was mixed with a 4 mM OPA‐NAC solution (prepared according to previous reports) (Shikanai et al, 2022) and reacted at room temperature for 150 s. After passing through a 0.22 μm MCE syringe filter, the samples were detected as follows: The mobile phase was a mixture of 100 mM phosphate buffer (pH 6.0)‐methanol containing 5 mg/L EDTA‐2Na (V/V, 82:18); the flow rate was set at 1.0 mL/min; the UV detector operated at 333 nm, and the column temperature was maintained at 30°C (Supporting Information S1: Figure S2D).…”

Section: Methodsmentioning

confidence: 99%

Efficient production of l‐glutathione by whole‐cell catalysis with ATP regeneration from adenosine

Zuo,

Zhao,

Zhang

et al. 2024

Biotech & Bioengineering

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Abstractl‐glutathione (GSH) is an important tripeptide compound with extensive applications in medicine, food additives, and cosmetics industries. In this work, an innovative whole‐cell catalytic strategy was developed to enhance GSH production by combining metabolic engineering of GSH biosynthetic pathways with an adenosine‐based adenosine triphosphate (ATP) regeneration system in Escherichia coli. Concretely, to enhance GSH production in E. coli, several genes associated with GSH and l‐cysteine degradation, as well as the branched metabolic flow, were deleted. Additionally, the GSH bifunctional synthase (GshFSA) and GSH ATP‐binding cassette exporter (CydDC) were overexpressed. Moreover, an adenosine‐based ATP regeneration system was first introduced into E. coli to enhance GSH biosynthesis without exogenous ATP additions. Through the optimization of whole‐cell catalytic conditions, the engineered strain GSH17‐FDC achieved an impressive GSH titer of 24.19 g/L only after 2 h reaction, with a nearly 100% (98.39%) conversion rate from the added l‐Cys. This work not only unveils a new platform for GSH production but also provides valuable insights for the production of other high‐value metabolites that rely on ATP consumption.

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Separation and detection of D-/L-serine by conventional HPLC

Cited by 2 publications

References 13 publications

Spectrophotometric Study of Excitatory Amino Acids with Ortho-Phthalaldehyde

Spectrophotometric Study of Excitatory Amino Acids with Ortho-Phthalaldehyde

Efficient production of l‐glutathione by whole‐cell catalysis with ATP regeneration from adenosine

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