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Method for detecting free amino acids in tea by using reversed-phase high-performance liquid chromatography

A reversed-phase high performance liquid phase, free amino acid technology, applied in the field of analytical chemistry, can solve the problem of not containing glutamine, and achieve the effect of simple sample pretreatment and low cost

Inactive Publication Date: 2015-06-03
ANHUI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method does not include the detection of glutamine

Method used

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  • Method for detecting free amino acids in tea by using reversed-phase high-performance liquid chromatography
  • Method for detecting free amino acids in tea by using reversed-phase high-performance liquid chromatography

Examples

Experimental program
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Effect test

Embodiment 1

[0030] The mensuration of free amino acid in embodiment 1 green tea

[0031] (1) Standard products: aspartic acid, serine, glutamic acid, glycine, glutamine, histidine, arginine, threonine, alanine, proline, tyrosine, cysteine, Valine, Methionine, Isoleucine, Lysine, Leucine, Phenylalanine, Theanine.

[0032] ⑵Sample pretreatment

[0033] Weigh 1.5g tea sample and soak in 200ml Watsons water, extract at 100°C for 45min, cool to room temperature, transfer to a 250ml volumetric flask after suction filtration, let stand to take the supernatant, filter with a 0.22um microporous membrane, seal the obtained filtrate Save it for the next step.

[0034] ⑶ Derivation of samples

[0035] The derivatizing agent is 6-aminoquinoline-N-hydroxysuccinimidyl formate (AQC), the derivatizing time of the derivatizing agent is 10 min, and the derivatizing temperature is 55°C. The free amino acids in the green tea sample were derivatized with the above derivation conditions; the derivation spec...

Embodiment 2

[0046] The mensuration of free amino acid in embodiment 2 black tea

[0047] (1) Standard products: aspartic acid, serine, glutamic acid, glycine, glutamine, histidine, arginine, threonine, alanine, proline, tyrosine, cysteine, Methionine, Isoleucine, Lysine, Leucine, Phenylalanine, Theanine.

[0048] ⑵Sample pretreatment

[0049] Weigh 1.5g tea sample and soak it in 200ml Watsons water, extract it at 100°C for 45min, cool to room temperature, transfer it to a 250ml volumetric flask after suction filtration, let it stand still to take the supernatant, filter it with a 0.22um microporous membrane filter, and seal the obtained filtrate Save it for the next step.

[0050] ⑶ Derivation of samples

[0051] The derivatization agent is 6-aminoquinoline-N-hydroxysuccinimidyl carboxylate (AQC), the derivatization time is 10min, and the derivatization temperature is 55°C. The free amino acids in the black tea sample were derivatized with the above derivation conditions, and the deri...

Embodiment 3

[0058] The mensuration of free amino acid in the oolong tea of ​​embodiment 3

[0059] (1) Standard products: aspartic acid, serine, glutamic acid, glycine, glutamine, histidine, arginine, threonine, alanine, proline, tyrosine, cysteine, Methionine, isoleucine, lysine, leucine, phenylalanine, and theanine were purchased from Sigma.

[0060] ⑵Sample pretreatment

[0061] Weigh 1.5g tea sample and soak it in 200ml Watsons water, extract it at 100°C for 45min, cool to room temperature, transfer it to a 250ml volumetric flask after suction filtration, let it stand still to take the supernatant, filter it with a 0.22um microporous membrane filter, and seal the obtained filtrate Save it for the next step.

[0062] ⑶ Derivation of samples

[0063] The derivatization agent is 6-aminoquinoline-N-hydroxysuccinimidyl carboxylate (AQC), the derivatization time is 10min, and the derivatization temperature is 55°C. The free amino acids in the oolong tea sample were derivatized with the ...

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Abstract

The invention relates to a method for detecting free amino acids in tea by using the reversed-phase high-performance liquid chromatography. The method comprises the following steps: drying and grinding fresh tea, and adding boiling water for digestion; after supernatant liquor is collected, repeatedly extracting and merging the supernatant liquor; after the supernatant liquor is sucked and filtered, carrying out precolumn derivatization for a tea soup sample with an AccQ.Tag derivatization reagent; optimizing chromatographic separation and detection conditions with HPLC and AccQ.Tag C18 special amino acid analysis columns, and finally separating and detecting 19 kinds of free amino acids such as glutamine. The method can simultaneously determine the contents of 19 kinds of free amino acids such as glutamine in the tea, makes up the vacancy that glutamine in the tea cannot be quickly and accurately analyzed based on the high-performance liquid chromatography qualitatively and quantitatively at present, and provides a new idea for researching the synthesis path of amino acids in the tea.

Description

technical field [0001] The invention belongs to the field of analytical chemistry, and in particular relates to a high-performance liquid chromatography (HPLC) method for simultaneously detecting the contents of 19 kinds of free amino acids in tea leaves. Background technique [0002] Amino acids are the most basic substances that constitute biological proteins, and are also important components of active peptides, enzymes, and other biologically active molecules in organisms, and are closely related to life activities. Tea leaves are rich in amino acids and thus provide the essential nutrients needed by the human body. Amino acids in tea are also one of the characteristic components that directly affect the quality of tea, and their content and composition are closely related to the taste and aroma of tea. Therefore, the analysis of amino acids in tea is of great significance to the quality identification and nutritional value evaluation of tea. [0003] At present, the c...

Claims

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Application Information

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IPC IPC(8): G01N30/89
Inventor 宛晓春李敏李大祥邰玉玲顾晨晨宗建法
Owner ANHUI AGRICULTURAL UNIVERSITY
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