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PAABSF column front derivatizing process of amino-acid

A pre-column derivatization, amino acid technology, applied in the field of analytical chemistry, can solve the problems of interference separation, poor reagent selectivity, unstable derivative products, etc., and achieve the effect of simple derivatization reaction conditions and less interference

Inactive Publication Date: 2006-08-30
YANGZHOU UNIV
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  • Application Information

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Problems solved by technology

For example: o-phthalaldehyde (OPA) [1] can only derivate primary amino acids, and the derivative products are unstable; fluorenylmethyl chloroformate (FMOC) [2] is a common reagent for derivatizing amine compounds, but it itself and Both produce fluorescence similar to derivatives, thereby interfering with separation; phenyl isothiocyanate (PITC) needs vacuum drying after derivatization with amino acids to remove excess reagents, and PITC consumes column life[3]; dimethylaminonaphthalenesulfonyl chloride (Dansyl-Cl)[4] derivatives are sensitive to light, and the selectivity of this reagent is poor
Para-acetaminobenzenesulfonyl fluoride (PAABS-F) was mainly used in organic synthesis in the past, and there is no report on pre-column derivatization

Method used

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  • PAABSF column front derivatizing process of amino-acid
  • PAABSF column front derivatizing process of amino-acid

Examples

Experimental program
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Effect test

Embodiment 1

[0010] Instruments and conditions: capillary electrophoresis apparatus, the total length of the capillary is 60cm, the effective column length is 50cm, and the inner diameter is 50μm. 13 kinds of amino acids (biochemical reagents, Sinopharm Group). SDS (biochemical reagent, Alfa Aesar).

[0011] Implementation steps: Quantitatively pipette 40 μL of each amino acid (0.05 mmol / L) into a 2 mL stoppered test tube, add 200 μL of PAABS-F ethanol solution (0.05 mmol / L) and 1 μL of methanol, mix well, and stopper tightly Put it in a constant temperature water bath at 35°C for 30min. Filter with a 0.22 μm membrane before injection.

Embodiment 2

[0013] Instruments and conditions: capillary electrophoresis apparatus, the total length of the capillary is 60cm, the effective column length is 50cm, and the inner diameter is 50μm. 13 kinds of amino acids (biochemical reagents, Sinopharm Group). SDS (biochemical reagent, Alfa Aesar).

[0014] Experimental procedure: Take 0.8mL of the pretreated serum sample, add 200μL of PAABS-F ethanol solution (0.05mmol / L) and 1μL of methanol, mix well, plug it tightly and put it in a constant temperature water bath at 35°C for 30min. Filter with a 0.22 μm membrane before injection.

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Abstract

The invention relates to a PAABS-F pre column deriving method of amino acid. The process includes the following steps: adding PAABS-F and little methanol into the amino acid sample after being pre-processed, mixing to equal, taking deriving reaction for 20-40 minutes under 25-45 degree centigrade, taking capillary electrophoresis or high efficiency liquid chromatography to take separating and detecting to gain the thickness of amino acid. The invention has simple and milt reaction condition, little disturbance and could directly take chromatographic analysis after reacting. The derivation result is stable and could place for seven days without avoiding light. The best deriving reaction temperature is 35 degree centigrade.'

Description

technical field [0001] The invention relates to a pre-column derivatization method of compounds, belonging to the technical field of analytical chemistry. Background technique [0002] Pre-column derivatization is a commonly used method to improve the detection sensitivity of amino acids, but there are more or less problems in the derivatization methods of amino acids reported so far. For example: o-phthalaldehyde (OPA) [1] can only derivate primary amino acids, and the derivative products are unstable; fluorenylmethyl chloroformate (FMOC) [2] is a common reagent for derivatizing amine compounds, but it itself and Both produce fluorescence similar to derivatives, thereby interfering with separation; phenyl isothiocyanate (PITC) needs vacuum drying after derivatization with amino acids to remove excess reagents, and PITC consumes column life[3]; dimethylaminonaphthalenesulfonyl chloride (Dansyl-Cl)[4] derivatives are sensitive to light, and the selectivity of this reagent is...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/14G01N30/02
Inventor 瞿其曙汤晓庆胡效亚刘尹陆晓
Owner YANGZHOU UNIV
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