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Metabonomics database establishment method and application of Chinese wolfberry tissues

A technology of metabolomics and database, which is applied in the field of establishment of metabolomics database of Lycium barbarum tissue, which can solve problems such as the influence of drug efficacy, affecting the application of Lycium barbarum, and difficult standardized production of traditional Chinese medicine.

Active Publication Date: 2020-10-20
WOLFBERRY ENG RES INST NINGXIA ACADEMY OF AGRI & FORESTRY SCI
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  • Description
  • Claims
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Problems solved by technology

[0003] However, the quality of wolfberry is different due to various methods of planting regions and planting techniques, and these differences will affect the application of wolfberry
For example, in the field of traditional Chinese medicine, due to the different quality of wolfberry, it will affect the efficacy of the medicine, which has become one of the factors that make it difficult to standardize the production of traditional Chinese medicine.

Method used

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  • Metabonomics database establishment method and application of Chinese wolfberry tissues
  • Metabonomics database establishment method and application of Chinese wolfberry tissues
  • Metabonomics database establishment method and application of Chinese wolfberry tissues

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Extraction method optimization experiments and data

[0035] Substance pretreatment:

[0036] The sample extraction process of this project is as follows:

[0037] Collect the leaves and fruit tissues of different varieties of Lycium barbarum, aggregate the leaves of different varieties, aggregate the fruits of different varieties, perform pretreatment respectively and add extraction reagents to extract leaf samples and fruit samples;

[0038] (1) The sample is vacuum freeze-dried;

[0039] (2) Utilize grinder to grind to powder;

[0040] (3) Weigh 40mg-50mg of powder and dissolve it in the extract solution with volume V=net sample weight (mg)×12μL / mg;

[0041] (4) Thoroughly mix the sample with the solvent, vortex each tube for 20 seconds, vortex once every half hour, vortex 5 times in total, and put it in a 4°C refrigerator overnight;

[0042] (5) After centrifugation (12,000 rpm, 10 minutes, 4°C), the supernatant was aspirated, the sample was filtered with a micr...

Embodiment 2

[0047] Mass Spectrometry Parameter Confirmation Experiment and Data

[0048] Chromatographic conditions:

[0049] (1) Chromatographic column: Waters ACQUITY UPLC HSS T3 C18 1.8μm, 2.1mm*100mm;

[0050] (2) Mobile phase: the aqueous phase is ultrapure water, and the organic phase is acetonitrile, all adding 0.1% acetic acid by volume percentage;

[0051] (3) Elution gradient: 0min water / acetonitrile (95:5V / V), 10.0min is 5:95V / V, 11.0min is 5:95V / V, 11.1min is 95:5V / V, 14.0min is 95 :5V / V;

[0052] (4) The flow rate is 0.4ml / min; the column temperature is 40°C; the injection volume is 2μl.

[0053] Mass spectrometry parameter screening test:

[0054] Electrospray ionization (ESI) temperature was 550°C, mass spectrometry voltage (+) 5500V, (-) 4500V, curtain gas (CUR) 35psi, collision-induced ionization (collision-activateddissociation, CAD) parameters were set to medium , the scanning mode is positive and negative ion scanning. In the triple quadrupole (QQQ), each ion pai...

Embodiment 3

[0060] Construction of Lycium barbarum leaf and Lycium barbarum fruit database:

[0061] After the preliminary screening of positive and negative ions and deduplication in Example 2, the following method was used to construct the database.

[0062] Step S1: Collect the leaves and fruit tissues of 20 different varieties of Lycium barbarum, group them together according to the leaves and fruits, and extract the leaf samples and fruit samples according to the method in Example 1 to obtain leaf sample extracts and fruit sample extracts;

[0063] Step S2: The extract obtained in the step S1 is detected by LC-MS / MS using UPLC-QTrap6500 with reference to the conditions confirmed in Example 2;

[0064] Step S3: Analyzing the chemical structure of each metabolite and locating ion chromatographic peaks;

[0065] Step S4: Build the database.

[0066] A total of 3,448 metabolite chromatographic peaks were detected. Among the 3,448 identified metabolites, the database was searched for kn...

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Abstract

The invention provides a metabonomics database establishment method and application of Chinese wolfberry tissues. The metabonomics database establishment method comprises the following steps: collecting leaves and fruit tissues of different varieties of Chinese wolfberry, summarizing the leaves of different varieties, summarizing the fruits of different varieties, respectively carrying out pretreatment, and adding an extraction reagent for extraction to obtain a leaf extracting solution and a fruit extracting solution; carrying out LC-MS / MS (Liquid Chromatography-Mass Spectrometry / Mass Spectrometry) detection on the extracting solution obtained in the step S1 and carrying out data analysis; analyzing the chemical structure of each metabolite and positioning an ion chromatographic peak; andbuilding a database. The method disclosed by the invention has the beneficial effects that (1) through mutual cooperation of a pretreatment method, a mass spectrometry method and a chromatographic method, a large amount of substances of the Chinese wolfberry leaves and the Chinese wolfberry fruits are rapidly identified, and a comprehensive metabolite database is constructed; (2) the metabolite database provides convenience for subsequent qualitative and quantitative analysis of the Chinese wolfberry metabolites; and (3) detection and determination of specific metabolites of the Chinese wolfberry leaves and the Chinese wolfberry fruits, so that a material basis is provided for subsequent metabolism research and development of Chinese wolfberry.

Description

technical field [0001] The invention belongs to the field of analytical chemistry, in particular to a method and application for establishing a metabolomics database of wolfberry tissue. Background technique [0002] Lycium barbarum is a widely used medicinal and edible crop, and its fruits and leaves have broad application prospects. [0003] However, the quality of wolfberry is different due to various methods of planting regions and planting techniques, and these differences will affect the application of wolfberry. For example, in the field of traditional Chinese medicine, due to the different quality of wolfberry, it will affect the efficacy of the medicine, which has become one of the factors that make it difficult to standardize the production of traditional Chinese medicine. [0004] Therefore, in order to better understand and identify the differences of different tissues of Lycium barbarum, maximize the utilization of its functional substances, and facilitate subs...

Claims

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Application Information

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IPC IPC(8): G16C20/70G16C20/90
CPCG16C20/70G16C20/90
Inventor 赵建华李浩霞尹跃安巍陈曦胡丽娟黄稳娥曹有龙
Owner WOLFBERRY ENG RES INST NINGXIA ACADEMY OF AGRI & FORESTRY SCI
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