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A method for simultaneous detection of carboxymethyllysine and carboxyethyllysine in green tea

A technology of carboxymethyllysine and carboxyethyllysine, which is applied in the field of analysis and detection, can solve problems affecting the applicability of the method, and achieve good stability

Inactive Publication Date: 2019-01-29
HANGZHOU TEA RES INST CHINA COOP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, LC-MS / MS analysis method has been reported to determine CML and CEL in food matrix. The pretreatment of this method is relatively simple, the detection speed is fast, the reproducibility is good, and the stability is good. However, different raw material matrices will affect the applicability of the method. sex

Method used

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  • A method for simultaneous detection of carboxymethyllysine and carboxyethyllysine in green tea
  • A method for simultaneous detection of carboxymethyllysine and carboxyethyllysine in green tea
  • A method for simultaneous detection of carboxymethyllysine and carboxyethyllysine in green tea

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] (1) Extraction: Weigh 50 mg tea powder sample into a 10 mL centrifuge tube, add 2 mL of n-hexane, shake vigorously for 3 min, then centrifuge at 10,000 rpm for 5 min, discard the solvent, and repeat the above steps twice. After degreasing with n-hexane, the sample was blown dry with nitrogen until it recovered to a powder state. Add 1mL Na to the sample 2 B 4 o 7 Buffer (0.2mol / L) and 0.5mL NaBH 4 (1mol / L, prepared with 0.1mol / L NaOH), adjust pH=9.2, and reduce at 4°C overnight (about 8h). Next, add 1 mL of chloroform / methanol (2:1, v / v), centrifuge at 10,000 rpm for 10 min, discard the supernatant; add 5 mL of HCI (6 mol / L) to the obtained precipitate, and place it in an oven at 110 ° C , Hydrolyzed for 12h. After the hydrolysis was completed, the hydrolyzed sample was taken out and cooled to room temperature (25°C), and the volume was adjusted to 10 mL with ultrapure water, then filtered with qualitative filter paper and the filtrate was collected.

[0035] (2) ...

Embodiment 2

[0044] (1) Extraction: Weigh 50 mg of tea powder (same as in Example 1) and place it in a 10 mL centrifuge tube, add 2 mL of n-hexane, vibrate vigorously for 3 min, then centrifuge at 5000 rpm for 10 min, discard the solvent, and repeat the above steps 3 times. After degreasing with n-hexane, the sample was blown dry with nitrogen until it recovered to a powder state. Add 1mL Na to the sample 2 B 4 o 7 Buffer (0.1mol / L) and 0.5mL NaBH 4 (0.8mol / L, prepared with 0.1mol / L NaOH), adjust pH=8.5, and reduce at 4°C for 10h. Next, add 1 mL of chloroform / methanol (1:1, v / v), centrifuge at 5000 rpm for 10 min, discard the supernatant; add 5 mL of HCI (4 mol / L) to the obtained precipitate, and place in an oven at 100 ° C , Hydrolyzed for 30h. After the hydrolysis was completed, the hydrolyzed sample was taken out and cooled to room temperature (25°C), and the volume was adjusted to 10 mL with ultrapure water, then filtered with qualitative filter paper and the filtrate was collecte...

Embodiment 3

[0054] (1) Extraction: Weigh 50 mg of tea powder (same as in Example 1) and place it in a 10 mL centrifuge tube, add 2 mL of n-hexane, vibrate vigorously for 3 min, then centrifuge at 12000 rpm for 4 min, discard the solvent, and repeat the above steps twice. After degreasing with n-hexane, the sample was blown dry with nitrogen until it recovered to a powder state. Add 1mL Na to the sample 2 B 4 o 7 Buffer (0.3mol / L) and 0.5mL NaBH 4 (1.2mol / L, prepared with 0.1mol / L NaOH), adjust pH=10, and reduce at 6°C for 2h. Next, add 1 mL of chloroform / methanol (3:1, v / v), centrifuge at 12000 rpm for 4 min, discard the supernatant; add 5 mL of HCI (8 mol / L) to the obtained precipitate, and place in an oven at 120 ° C , Hydrolyzed for 12h. After the hydrolysis was completed, the hydrolyzed sample was taken out and cooled to room temperature (25°C), and the volume was adjusted to 10 mL with ultrapure water, then filtered with qualitative filter paper and the filtrate was collected. ...

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Abstract

The invention relates to the technical field of analysis and detection, and provides a method for synchronously detecting carboxymethyl lysine and carboxyethyl lysine in green tea in order to solve the problem that an existing method for detecting CML and CEL will be affected by different raw material substrates. d4-CML and d4-CEL are used as interior labels, a multi-choice reaction monitoring mode is adopted, a quantitative analysis method for synchronously detecting CML and CEL in green tea is built through methodological argumentation, and a foundation is provided for monitoring and controlling AGEs in tea. The detection method is good in accuracy, reproducibility and stability, and applicable to practical detection of CML and CEL in green tea.

Description

technical field [0001] The invention relates to the technical field of analysis and detection, in particular to a method for synchronously detecting carboxymethyllysine and carboxyethyllysine in green tea. Background technique [0002] my country's tea garden area and tea output rank first in the world, and it is a big country in the tea industry. There are many types of tea, rich endoplasmic components, and various processing procedures. During the tea making process, the endoplasmic components will undergo complex chemical changes, and potential hazards such as advanced glycation end products (Advanced Glycation End Products, AGEs) will easily be produced. Tea processing Process safety has become a new focus in the field of tea quality and safety. AGEs, one of the products of Maillard reaction, are a class of compounds with complex structures, which have the characteristics of irreversibility, cross-linking, not easy to be degraded, structural heterogeneity, and enzyme st...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06
Inventor 王彬刘婧田野曾丽珍刘青茹周春江蒋玉兰唐靖邸胜达
Owner HANGZHOU TEA RES INST CHINA COOP
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