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Method for rapidly determining L-glutamic acid in food

A technology for rapid determination of glutamic acid, applied in the direction of measuring devices, biological testing, material inspection products, etc., can solve the problems of expensive instruments and consumables, failure to achieve high throughput, and non-specificity, etc., to achieve simple operation, The instrument price and detection reagent are low in cost and easy to promote.

Inactive Publication Date: 2016-12-21
CENT TESTING INT GRP CO LTD
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  • Summary
  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

Among them, the optical rotation method, the ultraviolet spectrophotometer method, the neutralization titration method, and the ninhydrin method are not specific, and are only suitable for the detection of glutamic acid in monosodium glutamate with a single component; -Glutamic acid decarboxylase control has specificity, but the operation is more complicated, and it is easy to introduce errors if you don't pay attention; the amino acid automatic analyzer method is an improved ninhydrin method, which is suitable for the analysis of all amino acids, and is easy and accurate to operate Good performance, but the instruments and consumables are expensive, and only one sample can be detected at a time, failing to achieve high throughput

Method used

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  • Method for rapidly determining L-glutamic acid in food
  • Method for rapidly determining L-glutamic acid in food

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preparation example Construction

[0030] 1. Preparation of sample detection solution:

[0031] Solid or semi-solid samples are crushed or minced with a grinder or a meat grinder, and 50 g is put into a homogenizing cup, and 100 mL of 1.0 mol / L dilute perchloric acid solution is added for homogeneous extraction for 5 minutes (Note: the extraction method The resulting L-glutamic acid is free L-glutamic acid, if you want the extract to include L-glutamic acid that constitutes the protein structure, you need to increase the proteolysis step), transfer the mixture to a 50mL centrifuge tube, and centrifuge at 1000g After 5 minutes, carefully remove the floating matter in the upper layer, take the middle clear liquid, and filter it with a medium-speed filter paper (the liquid sample does not need the above pretreatment, just filter it directly), take an appropriate amount of filtrate, and adjust the pH with 2mol / L potassium hydroxide solution to 8.0 to 9.0, and diluted to a certain volume, as a sample detection solut...

Embodiment 1

[0047] 1. Preparation of sample detection solution

[0048] Weigh 2g of soy sauce sample into a beaker, accurate to 0.01g, dilute with water, adjust the pH to 8.6 with 2.0mol / L potassium hydroxide solution, transfer to a 500mL volumetric flask, and set the volume to the mark, filter to make the soy sauce sample detectable The concentration of liquid L-glutamic acid is controlled within the range of 0.006g / L to 0.06g / L.

[0049] 2. Preparation of Assay Reagents

[0050] Reagent A1: Weigh 1.488g of triethanolamine phosphate, dipotassium hydrogen phosphate (K 2 HPO 4 ) 0.172g, potassium dihydrogen phosphate (KH 2 PO 4) 1.4mg, dissolved in water, adjusted the pH to 8.6 with 2mol / L potassium hydroxide solution, added 0.5g of Triton X-100, dilute to 100mL, and made triethanolamine phosphate buffer solution with pH 8.6 . The solution is stored at 2°C to 8°C, and the shelf life is 2 months.

[0051] Reagent A2: Weigh diaphorase freeze-dried powder (enzyme activity unit is 25U / m...

Embodiment 2

[0067] 1. Preparation of sample detection solution

[0068] Cut about 100g of chicken into small pieces, grind it with a meat grinder, weigh 50g, accurate to 0.01g, put it into a homogenizing cup, and add 100mL of 1.0mol / L dilute perchloric acid at 0°C to the homogenizing cup solution, homogenized extraction for 5 minutes (note: the L-glutamic acid obtained by this extraction method is free L-glutamic acid, if you want the extract to include L-glutamic acid that constitutes the protein structure, you need to increase the proteolysis step), Transfer the homogenized mixture to a 50mL centrifuge tube, centrifuge at 2000g for 5 minutes, carefully remove the upper lipid film, take the middle supernatant, and filter.

[0069] Draw 10mL of the filtrate into a beaker, dilute with water, adjust the pH to 8.6 with 2.0mol / L potassium hydroxide solution, transfer it to a 100mL volumetric flask, and set the volume to the mark, so that the chicken sample detection solution L-glutamic acid c...

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Abstract

The present invention relates to a method for rapidly determining the L-glutamic acid in food. The method comprises: grinding or mincing a solid or semi-solid sample by using a suitable tool, weighing a proper amount of the grinded or minced sample, adding an appropriate solvent, homogenizing, carrying out centrifuged separation, and filtering to obtain a free L-glutamic acid extraction liquid, or carrying out protein hydrolysis to obtain the L-glutamic acid extraction liquid constituting the protein structure (the liquid sample is directly filtered without the pretreatment); properly diluting the sample extraction liquid, and adjusting the pH value to 8.6; reducing nicotinamide adenine dinucleotide (NAD<+>) into nicotinamide adenine dinucleotide (NADH) in the presence of glutamate dehydrogenase(GIDH) through the L-glutamic acid of the sample detection liquid, and carrying out a reaction on the NADH and iodonitrotrtrazolium chloride (INT) under the effect of diaphorase to generate formazan; and determining the absorption peak of the formazan at 490 nm by using a light-absorption microplate reader, and calculating the L-glutamic acid content in the sample according to the absorption value of the formazan by using an external standard method. According to the present invention, the detection method has characteristics of simple operation, good accuracy, fastness, high throughput, easy popularization, and the like.

Description

technical field [0001] The invention relates to a method for rapidly determining L-glutamic acid in food, in particular to a redox reaction principle based on glutamic acid dehydrogenase and diaphorase two-way control, combined with a light absorption microplate reader to rapidly detect L-glutamic acid - Glutamate method. Background technique [0002] L-glutamic acid widely exists in the organisms of animals and plants, and is a naturally occurring nutrient in food. At present, there are many detection methods for L-glutamic acid, including polarimetry, ultraviolet spectrophotometry, neutralization titration, ninhydrin method, Valsalva respiration method, and amino acid automatic instrument method. Among them, the optical rotation method, the ultraviolet spectrophotometer method, the neutralization titration method, and the ninhydrin method are not specific, and are only suitable for the detection of glutamic acid in monosodium glutamate with a single component; -Glutamic ...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6812
Inventor 缪素娜戴煦董宁刘攀超刘泽华
Owner CENT TESTING INT GRP CO LTD
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