CN102095775A - Method for efficiently separating and detecting theanine enantiomer by capillary electrophoresis - Google Patents

Abstract

The invention discloses a method for efficiently separating and detecting theanine enantiomer by capillary electrophoresis, belonging to the technical field of separation and detection of enantiomers. The method determines the chiral separation environment required by separation of theanine, a borate buffer liquid system with pH value of 10 to 12 is selected, and a chiral selecting reagent is added in the buffer liquid. A theanine sample is prepared, the concentration is 0.1 to 2.0mg/mL, and the theanine enantiomer is separated on baseline in a condition of 15-30kV separation voltage of capillary electrophoresis. The method has good separating efficiency, no pollution and low cost, and is applicable to analysis and detection of theanine enantiomer.

Description

A kind of high performance capillary electrophoresis splits the method that detects the theanine enantiomorph

Technical field

A kind of high performance capillary electrophoresis splits the method that detects the theanine enantiomorph, belongs to the fractionation and the detection technique field of enantiomorph.

Background technology

The fractionation of enantiomorph is separating one of work that once was considered to the most difficult on the subject with detecting.Traditional method for splitting has fractional crystallization, microorganism and enzyme digestion etc., but these methods all are difficult to carry out the separation and the mensuration of trace.The high performance capillary electrophoresis technology is applied to enantiomorph split to detect development very rapid in recent years, and this technology has following advantage: 1. do not have the post pollution problem substantially; 2. experimental cost is low, consumes few; 3. analysis speed is fast, the separation efficiency height; 4. operator scheme is many.Based on above advantage, the chiral separation of Capillary Electrophoresis is own through becoming an extremely active popular domain.Need when high performance capillary electrophoresis is used for chiral resolution to add chiral selector, make up chirality and select environment, utilize it to separate with difference between the effects between enantiomorph at buffer solution.At present domestic and foreign literature does not also adopt the report of capillary electrophoresis technique separation detection theanine enantiomorph, and the present invention utilizes this technology to set up the analyzing and testing that a kind of good separating effect, highly sensitive analytical approach are applied to the theanine enantiomorph.

Summary of the invention

Purpose of the present invention: provide a kind of high performance capillary electrophoresis to split the method that detects the theanine enantiomorph, with beta-schardinger dextrin-and derivant thereof is chiral selector, make up chirality and select environment, separate and analyze the theanine enantiomorph, for the research and development of theanine provide technical guarantee.

Technical scheme of the present invention: a kind of high performance capillary electrophoresis splits the method that detects the theanine enantiomorph, and step is as follows:

(1) selecting the pH value for use is 10 ~ 12 borate buffer solution systems, adding concentration in this borate buffer solution system is 20 ~ 30mmol/L chirality selective reagent, ultrasonic degas 30-40min, with 0.45 μ m filtering with microporous membrane, filtering finishes continues ultrasonic degas 20 ~ 30min;

(2) with deionized water configuration theanine sample, concentration is 0.1 ~ 2.0mg/mL;

(3) regulating the Capillary Electrophoresis parameter is: separation voltage is 15 ~ 30KV, 20 ~ 30 ℃ of capillary temperatures, pressure sample introduction 3kPa * 2s, ultraviolet detection wavelength 220 ~ 280 nm; Positive pole advances the theanine sample, and negative pole detects;

(4) under described Capillary Electrophoresis parameter, make the theanine enantiomorph reach baseline separation;

Described chirality selective reagent is a beta-schardinger dextrin-, hydroxyethyl-, HP-, sulfonylation-beta-schardinger dextrin-or glucose group-beta-cyclodextrin.

Beneficial effect of the present invention: the present invention compares with traditional chiral separation has good separating effect, pollution-free, low cost, advantage such as analysis time is short.

Description of drawings

The electrophoretogram of Fig. 1 embodiment 1 theanine enantiomorph.

The electrophoretogram of Fig. 2 embodiment 2 theanine enantiomorphs.

The electrophoretogram of Fig. 3 embodiment 3 theanine enantiomorphs.

Embodiment

Below provide embodiment, the invention will be further described.

Embodiment 1

Adopting the pH value is that 11 borate solutions are damping fluid, adds 20 mmol/L beta-schardinger dextrin-s as the chirality selective reagent, crosses 0.45 μ m miillpore filter, ultrasonic degas 20min again behind the ultrasonic degas 40min of damping fluid elder generation.With deionized water configuration theanine sample, concentration is 0.1mg/mL, and the instrument that experiment is used is the P/ACE MDQ of Beckman company capillary electrophoresis system, and the Capillary Electrophoresis conditional parameter is: separation voltage is 20KV; 20 ℃ of capillary temperatures; Pressure sample introduction 3kPa * 2s; Ultraviolet detection wavelength 280 nm; Positive pole advances the theanine sample, and negative pole detects.The electrophoretogram of theanine enantiomorph is seen Fig. 1.

Embodiment 2

Adopting the pH value is that 12 borate solutions are damping fluid, adds 30 mmol/L hydroxyethyl-s as the chirality selective reagent, crosses 0.45 μ m miillpore filter, ultrasonic degas 20min again behind the ultrasonic degas 30min of damping fluid elder generation.With deionized water configuration theanine sample, concentration is 1.0mg/mL, and the instrument that experiment is used is the P/ACE MDQ of Beckman company capillary electrophoresis system, and the Capillary Electrophoresis conditional parameter is: separation voltage is 30KV; 25 ℃ of capillary temperatures; Pressure sample introduction 3kPa * 2s; Ultraviolet detection wavelength 280 nm; Positive pole advances the theanine sample, and negative pole detects.The electrophoretogram of theanine enantiomorph is seen Fig. 2.

Embodiment 3

Adopting the pH value is that 12 borate solutions are damping fluid, adds 25 mmol/L glucose group-beta-cyclodextrins as the chirality selective reagent, crosses 0.45 μ m miillpore filter, ultrasonic degas 20min again behind the ultrasonic degas 30min of damping fluid elder generation.With deionized water configuration theanine sample, concentration is 2.0mg/mL, and the instrument that experiment is used is the P/ACE MDQ of Beckman company capillary electrophoresis system, and the Capillary Electrophoresis conditional parameter is with embodiment 1.The electrophoretogram of theanine enantiomorph is seen Fig. 3.

Claims (1)

1. a high performance capillary electrophoresis splits the method that detects the theanine enantiomorph, it is characterized in that step is as follows:

(1) selecting the pH value for use is 10 ~ 12 borate buffer solution systems, adding concentration in this borate buffer solution system is 20 ~ 30mmol/L chirality selective reagent, ultrasonic degas 30-40min, with 0.45 μ m filtering with microporous membrane, filtering finishes continues ultrasonic degas 20 ~ 30min;

(2) with deionized water configuration theanine sample, concentration is 0.1 ~ 2.0mg/mL;

(3) regulating the Capillary Electrophoresis parameter is: separation voltage is 15 ~ 30KV, 20 ~ 30 ℃ of capillary temperatures, pressure sample introduction 3kPa * 2s, ultraviolet detection wavelength 220 ~ 280 nm; Positive pole advances the theanine sample, and negative pole detects;

(4) under described Capillary Electrophoresis parameter, make the theanine enantiomorph reach baseline separation;

Described chirality selective reagent is a beta-schardinger dextrin-, hydroxyethyl-, HP-, sulfonylation-beta-schardinger dextrin-or glucose group-beta-cyclodextrin.

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