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Method for detecting thaumatin in beverage

A detection method and beverage technology, which can be used in measurement devices, material separation, and analysis of materials, etc., can solve the problems of less reporting of detection methods, and achieve the effect of reliable results.

Inactive Publication Date: 2019-04-26
南通市产品质量监督检验所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, there is no detection method standard for thaumatin in China, and there are very few reports on its detection method, only the research on the detection method of sweeteners including thaumatin by using high performance liquid chromatography-evaporative light detector

Method used

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  • Method for detecting thaumatin in beverage

Examples

Experimental program
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Effect test

Embodiment 1

[0030] This embodiment provides a case of utilizing ultra-high performance liquid chromatography to analyze thaumatin in carbonated beverages, and the specific steps are as follows:

[0031] (1) Chromatographic conditions: Agilent 1290 ultra-high performance liquid chromatography; Chromatographic column: 300SB-C18, RRHD 1.8μm, 100mm×2.1mm(i,d); Mobile phase: acetonitrile, 0.1% trifluoroacetic acid aqueous solution; Flow rate: 0.3mL / min; injection volume: 20μL; column temperature: 25°C. Gradient elution: 0-3min, acetonitrile 10%-0.1% trifluoroacetic acid aqueous solution 90%; 3-15min, acetonitrile (10%-100%), 0.1% trifluoroacetic acid aqueous solution (90%-0%); 15.1- 20min, acetonitrile (100%); 20.1-25min, acetonitrile 10%-0.1% trifluoroacetic acid aqueous solution 90%; detector: DAD, detection wavelength 278.2nm, while scanning at 190-400nm wavelength;

[0032] (2) Preparation of standard curve: Dilute thaumatin with water to concentrations of 10 mg / L, 25mg / L, 50mg / L, 100mg / L...

Embodiment 2

[0035]This embodiment provides a case of using ultra-high performance liquid chromatography to analyze thaumatin in tea beverages. The specific steps are as follows:

[0036] (1) Chromatographic conditions: Agilent 1290 ultra-high performance liquid chromatography; Chromatographic column: 300SB-C18, RRHD 1.8μm, 100mm×2.1mm(i,d); Mobile phase: acetonitrile, 0.1% trifluoroacetic acid aqueous solution; Flow rate: 0.3mL / min; injection volume: 20μL; column temperature: 25°C. Gradient elution: 0-3min, acetonitrile 10%-0.1% trifluoroacetic acid aqueous solution 90%; 3-15min, acetonitrile (10%-100%), 0.1% trifluoroacetic acid aqueous solution (90%-0%); 15.1- 20min, acetonitrile (100%); 20.1-25min, acetonitrile 10%-0.1% trifluoroacetic acid aqueous solution 90%; detector: DAD, detection wavelength 278.2nm, while scanning at 190-400nm wavelength;

[0037] (2) Preparation of standard curve: Dilute thaumatin with water to concentrations of 10 mg / L, 25mg / L, 50mg / L, 100mg / L, 200mg / L, 500mg...

Embodiment 3

[0040] This embodiment provides a case of utilizing SDS-PAGE method to analyze thaumatin, and the specific steps are as follows:

[0041] (1) Sample treatment: Dilute the thaumatin standard with water to a concentration of 10mg / L, 25 mg / L, 50mg / L, 100mg / L, 200mg / L, 500mg / L, 1000mg / L series of standard solutions, respectively Take 80 μL of thaumatin standard solution and the spiked sample to be tested, add 20 μL of 5X protein loading buffer, and heat-treat in boiling water for 5 minutes;

[0042] (2) SDS-PAGE electrophoresis: electrophoresis was performed using a separating gel with a concentration of 12.0%, the loading volume of thaumatin standard solution and sample to be tested was 60 μL, the electrophoresis conditions were 135V, 50min, stained with Coomassie brilliant blue, and decolorized with decolorization solution; As a reference, the molecular weight range of the protein standard sample is 14.4kD~98kD, and the sample volume is 10μL; among them, the SDS-PAGE results of ...

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Abstract

The invention relates to a method for detecting thaumatin in a beverage, in particular to a method for analyzing thaumatin in the beverage through ultra-high performance liquid chromatography and an SDS-PAGE method. According to ultra-high performance liquid chromatography, after the beverage is subjected to ultrafiltration treatment, chromatographic filler large in pore size serves as a separation medium, a DAD serves as a detector, liquid chromatographic analysis is carried out, and qualitative and quantitative detection is performed. According to the SDS-PAGE method, 12.5% separation glue is used, coomassie brilliant blue is used for dyeing, qualitative and semi-quantitative analysis of thaumatin can be achieved, the sensitivity meets standard limit use requirements, and a way is provided for further analysis and verification. The method has the advantages that the method for analyzing thaumatin in the beverage is provided, the sensitivity meets the requirement for the limited amount of thaumatin, and the chromatographic method and an electrophoresis method can prove each other so as to confirm a sample.

Description

technical field [0001] The invention belongs to the technical field of physical and chemical testing of food, in particular to a detection method of thaumatin in beverages. Background technique [0002] Thaumatin (also translated as Suomatweet, Thaumatin, Thaumatin) is a super-sweet protein extracted from the fruit of the natural plant Thaurnatococcusdaniellii, which has similar properties to other plant-derived proteins such as Monellin, fairy Curculin, Mabinlin, Miraculin, Pentadin, Brazzein, Neoculin, etc. are collectively called sweet proteins. Compared with traditional carbohydrate sugars or chemically synthesized sweeteners, these natural sweet proteins generally have the advantages of high sweetness, low calories, good taste, safety and non-toxicity, etc. It is considered to be a very promising food additive because of the various amino acids it needs and the characteristics that it will not cause blood sugar rise and tooth decay. Studies have found that thaumatin i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N27/447
CPCG01N27/447G01N30/02
Inventor 邵彪周小兰陈刚王琳琳
Owner 南通市产品质量监督检验所
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