Rapid resolution liquid chromatography (RRLC) analysis of amino acids using pre-column derivatization

Zhang, Xiaoli; Zhao, Tong; Cheng, Ting; Liu, Xiaoyan; Zhang, Haixia

doi:10.1016/j.jchromb.2012.08.030

Cited by 41 publications

(24 citation statements)

References 24 publications

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“…S1); however, other amino acids such as GSH, Hcy, Arg, Gly, Met, Pro, Ser, Thr, Trp, Val and L-cystine could not react with CDNB at pH 6.8. As well known, in alkaline environment, CDNB can react with amino group of amino acids and thereby produce yellow conjugates, which has been used for the detection of amino acids (Sanger, 1945;Zhang et al, 2012). In our study, under the near neutral conditions (pH 6.8), it was firstly reported that CDNB could specifically conjugate with L-cysteine to produce the yellow conjugate DNPC, owing to the highly selective reaction between the thiol of L-cysteine and CDNB, as shown in Scheme 1.…”

Section: Resultsmentioning

confidence: 99%

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Ultrasensitive colorimetric detection of Cu2+ ion based on catalytic oxidation of l-cysteine

Yin

Wang

et al. 2015

Biosensors and Bioelectronics

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Section: Resultsmentioning

confidence: 99%

“…It is well known that L-cysteine is usually detected by derivatization methods through high performance liquid chromatography (Lindroth and Mopper, 1979;Zhang et al, 2012). However, derivatization processes are often quite complicated and easily interfered.…”

Section: Conjugation Of L-cysteine With Cdnbmentioning

confidence: 99%

Ultrasensitive colorimetric detection of Cu2+ ion based on catalytic oxidation of l-cysteine

Yin

Wang

et al. 2015

Biosensors and Bioelectronics

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“…The reagent 2,4-dinitrofluorobenzene (DNFB) is a versatile and highly efficient precolumn reagent that was found by Sanger [12]. The N-2,4-dinitrophenylated amine derivatives have been shown to be stable under ambient laboratory conditions [13,14]. This current work describes a rapid and accurate HPLC method, derivatized by DNFB, to quantify the intracellular Asp, Glu, Gly, and GABA content of primary cultured neurons, and the used culture medium.…”

Section: Introductionmentioning

confidence: 99%

Detection of four different amino acid neurotransmitters in cultured rat neurons and the culture medium by precolumn derivatization high-performance liquid chromatography

Zhang

Wang

et al. 2016

NeuroReport

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We validated and used a high-performance liquid chromatography procedure for the determination of four different amino acid neurotransmitters in cultured rat neurons and used culture medium. Samples were derivatized using 2,4-dinitrofluorobenzene and the amino acids were separated on a C18 column. The method yielded good reproducibility and sensitivity for the quantification of the four free amino acid neurotransmitters, with average recovery factors of 80.25-118.43%, an intraday precision of 0.09-0.17%, and an interday precision of 0.62-0.74%. The assay method can be readily utilized as a precise, sensitive, and highly accurate method for the determination of concentrations of the four amino acid neurotransmitters in cultured rat neurons and used culture medium. Using the described methods, we found that the aspartate, glutamic acid, glycine, and γ-aminobutyric acid concentrations (µmol/g protein) in cultured rat neurons were 25.23±0.81, 35.16±0.32, 77.56±4.51, and 62.87±3.12, respectively, whereas their concentrations (µM) in the used culture medium were 18.18±0.82, 24.27±1.01, 107.18±9.56, and 35.78±2.98, respectively.

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“…These compounds are proteins' building blocks that can also act as precursors for biosynthesis of many important biological and physiological substances. They also have an essential role in energy metabolism, neurotransmission and fat transport (1). Determining the concentration of amino acids is necessary to evaluate the pathophysiological processes of diseases (2).…”

Section: Introductionmentioning

confidence: 99%

Analysis of Plasma Amino Acids Using RP-HPLC and Pre-Column Derivatization with OPA/3-MPA

et al. 2016

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Background and Objective: Measurement of amino acids is an important tool for metabolic studies and evaluation of patients' clinical condition. The aim of this study was to analyze the plasma amino acids using reverse phase high performance liquid chromatography techniques (RP-HPLC) with pre-column derivatization by o-phthalaldehyde (OPA) in combination with 3-mercaptopropionic acid (3-MPA). Methods: Overall, 107 neonates and babies suspected of having metabolic disorder were enrolled in this study. The level of amino acids in plasma samples was analyzed within 65 minutes by HPLC with pre-column derivatization by OPA/3-MPA. This was a gradient RP-HPLC method that was performed using two solvents with a ratio of methanol and sodium acetate. L-norvaline internal standard was used as the reference peak for amino acids. Standard mixture of amino acids was used to determine the concentrations of amino acids. Results: According to the values of coefficient of variation obtained for each amino acid, the results indicated a good chromatographic separation of amino acids by this method. The use of OPA/3-MPA derivative reagent increased the efficiency and resolution of amino acids chromatographic separation. Conclusion: Due to simple preparation and accurate assessment, determination of plasma amino acids using OPA/3-MPA derivatives and RP-HPLC is a suitable method in many clinical samples.

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Rapid resolution liquid chromatography (RRLC) analysis of amino acids using pre-column derivatization

Cited by 41 publications

References 24 publications

Ultrasensitive colorimetric detection of Cu2+ ion based on catalytic oxidation of l-cysteine

Ultrasensitive colorimetric detection of Cu2+ ion based on catalytic oxidation of l-cysteine

Detection of four different amino acid neurotransmitters in cultured rat neurons and the culture medium by precolumn derivatization high-performance liquid chromatography

Analysis of Plasma Amino Acids Using RP-HPLC and Pre-Column Derivatization with OPA/3-MPA

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