Indirect Enantioseparation of Amino Acids by CE Using Automated In-Capillary Derivatization with ortho-Phthalaldehyde and N-Acetyl-l-Cysteine

Kühnreich, Raphael; Holzgrabe, Ulrike

doi:10.1007/s10337-016-3122-0

Cited by 16 publications

(15 citation statements)

References 42 publications

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“…There were 12 papers on in‐capillary derivatisation . The derivatisation was performed by adding the reagents to the separation buffer or by zone injections of reagents and sample.…”

Section: Derivatisation Modesmentioning

confidence: 99%

“…The three zone injection in combination with electrokinetic mixing was used for the analysis of racemic amino acids, glucose, and the drug 2‐mercaptoethanesulfonate. Kühnreich and Holzgrabe performed OPA labelling of 19 amino acids by sandwiching the sample between derivatisation mixtures . The derivatisation mixture consisted of OPA in 20 mM borate at pH 10 and 0.3 mM N ‐acetyl‐l‐cysteine (NAC).…”

Section: Derivatisation Modesmentioning

confidence: 99%

“…This is often the case for fluorescence detection where native fluorescence is rather uncommon and only the labelled species provide a strong signal. There were 24 articles on LIF , 16 on UV , 3 on electrochemical detection , and 4 on MS . In MS detection, derivatisation was used to improve the separation (see Section ) and to enable MS detection in favourable positive ion ESI mode .…”

Section: Derivatisation For Detectionmentioning

confidence: 99%

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Derivatisation for separation and detection in capillary electrophoresis (2015–2017)

Wuethrich

Quirino

2017

Electrophoresis

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Derivatisation is an integrated part of many analytical workflows to enable separation and detection of the analytes. In CE, derivatisation is adapted in the four modes of pre-capillary, in-line, in-capillary, and post-capillary derivatisation. In this review, we discuss the progress in derivatisation from February 2015 to May 2017 from multiple points of view including sections about the derivatisation modes, derivatisation to improve the analyte separation and analyte detection. The advancements in derivatisation procedures, novel reagents, and applications are covered. A table summarising the 46 reviewed articles with information about analyte, sample, derivatisation route, CE method and method sensitivity is provided.

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“…There were 12 papers on in‐capillary derivatisation . The derivatisation was performed by adding the reagents to the separation buffer or by zone injections of reagents and sample.…”

Section: Derivatisation Modesmentioning

confidence: 99%

Section: Derivatisation Modesmentioning

confidence: 99%

Section: Derivatisation For Detectionmentioning

confidence: 99%

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Derivatisation for separation and detection in capillary electrophoresis (2015–2017)

Wuethrich

Quirino

2017

Electrophoresis

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“…Here, we study and optimize the automated precolumn derivatization procedure that takes place inside the needle of an autosampler, which is crucial for the quantitative conversion of the AAs into its derivatives and especially for a low carryover effect. A method with a similar approach by means of capillary electrophoresis has been published previously by the authors . It aims to derivatize the AAs using OPA and either N‐acetyl‐L‐cysteine (NAC) or N‐isobutyryl‐L‐cysteine (NIBLC) as thiol.…”

Section: Introductionmentioning

confidence: 99%

“…A method with a similar approach by means of capillary electrophoresis has been published previously by the authors. 19 It aims to derivatize the AAs using OPA and either N-acetyl-L-cysteine (NAC) or N-isobutyryl-L-cysteine (NIBLC) as thiol. The principle of the derivatization process is to draw consecutively various reagent solutions in a specific order into the needle of the autosampler.…”

mentioning

confidence: 99%

High‐performance liquid chromatography evaluation of the enantiomeric purity of amino acids by means of automated precolumn derivatization with ortho‐phthalaldehyde and chiral thiols

Kühnreich

Holzgrabe

2016

Chirality

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The use of ortho-phthalaldehyde (OPA) for the derivatization of amino acids (AA) is well known. It enables the separation of the derivatives on common reversed phase columns and improves the sensitivity with fluorescence detection. With the use of a chiral thiol an indirect enantioseparation of chiral amines and AAs is feasible. The major drawback of the OPA-derivatization is the poor stability of the products. Here, a method with an in-needle derivatization procedure is optimized to facilitate a quantitative conversion of the AA with OPA and the chiral thiols N-acetyl-L-cysteine or N-isobutyryl-L-cysteine, followed by a subsequent analysis, eluding the stability issue. Both enantiomers of a single AA were separated as OPA-derivatives with a pentafluorophenyl column and a gradient program consisting of 50 mM sodium acetate buffer pH = 5.0 and acetonitrile. Fluorescence detection is commonly used to achieve sufficient sensitivity. In this study, the enantiomeric impurity of an AA can be detected indirectly with common UV spectrophotometric detection with a limit of quantitation of 0.04%. Seventeen different L-AAs were tested and the amount of D-AA for each individual AA was calculated by means of area normalization, which ranged from not detectable up to 4.29%. The recovery of the minor enantiomer of L- and D-AA was demonstrated for three AAs at a 0.04% level and ranged between 92.3 and 113.3%, with the relative standard deviation between 1.7 and 8.2%.

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Target‐Fluorophore Binding

2023

Fluorescent Dye Labels and Stains

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Indirect Enantioseparation of Amino Acids by CE Using Automated In-Capillary Derivatization with ortho-Phthalaldehyde and N-Acetyl-l-Cysteine

Cited by 16 publications

References 42 publications

Derivatisation for separation and detection in capillary electrophoresis (2015–2017)

Derivatisation for separation and detection in capillary electrophoresis (2015–2017)

High‐performance liquid chromatography evaluation of the enantiomeric purity of amino acids by means of automated precolumn derivatization with ortho‐phthalaldehyde and chiral thiols

Target‐Fluorophore Binding

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