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Method for identifying Chinese angelica and its adulterants by utilizing ITS sequence

A technique for mixing counterfeit products and angelica, applied in the field of identification of Chinese medicinal materials source varieties, can solve problems such as difficult identification, and achieve the effects of remarkable results, strong practicability, and easy operation.

Inactive Publication Date: 2014-08-27
INST OF MEDICINAL PLANT DEV CHINESE ACADEMY OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The counterfeit products of Angelica sinensis on the market mainly include Angelica sinensis, Soil Angelica, Ligusticum edulis, and Peucedanum chinensis, etc. Their shapes are similar, and it is difficult to identify them when they are mixed with genuine angelica.

Method used

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  • Method for identifying Chinese angelica and its adulterants by utilizing ITS sequence
  • Method for identifying Chinese angelica and its adulterants by utilizing ITS sequence
  • Method for identifying Chinese angelica and its adulterants by utilizing ITS sequence

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1 Angelica sinensis and its counterfeit product GenBank data analysis

[0023] Download the ITS sequence data of Angelica sinensis and its counterfeit products from GenBank, use the bioinformatics software CodonCode Aligner and MEGA5 for comparative analysis, and establish a Neighbor Joining Tree (NJ tree), the results are as follows Figure 5 As shown, the ITS sequence can well separate Angelica sinensis and its counterfeit products;

Embodiment 2

[0024] Example 2 Using ITS sequence to identify Angelica sinensis and its counterfeit products

[0025] 1. Sample source

[0026] Angelica sinensis and its counterfeit products include Angelica sinensis, Qingui, Duhuo, Ligusticum ligusticum, Ligusticum ligusticum Xinjiang, Angelica sinensis, Peucedanum chinensis, etc., which were collected from Gansu, Sichuan, Yunnan, Shaanxi, Hunan, Beijing and other places. Peucedanum decursiva (Miq.) Franch.&Sav.), Angelica sinensis (Levisticum officinale W.D.J.Koch), Ligusticum jeholense Nakai et Kitag., Conioselinum vaginatum (Spreng.) Thell. , Angelica pubescens Maxim.f.biserrata Shan et Yuan.

[0027] 2. DNA extraction

[0028] Take about 30 mg of the above samples, add sterilized small steel balls, grind them into powder with a sample grinder, and use the kit method to extract DNA. The kit was purchased from TianGen Company, and the model was Plant Genomic DNA Extraction Kit (spin column type) 200preps.

[0029] 3. PCR amplificatio...

Embodiment 3

[0043] Example 3 Using ITS sequence to identify commercially available Angelica sinensis samples

[0044] 1. Sample source

[0045] The 10 samples of Angelica sinensis used were collected from the traditional Chinese medicine Angelica sinensis sold in the market.

[0046] 2. DNA extraction

[0047] Take about 30 mg of the above samples, add sterilized small steel balls, grind them into powder with a sample grinder, and use the kit method to extract DNA. The kit was purchased from TianGen Company, and the model was Plant Genomic DNA Extraction Kit (spin column type) 200preps.

[0048] 3. PCR amplification

[0049] The PCR reaction system is calculated as 25 μl:

[0050] wxya 2 O: 8.5 μl

[0051] 2×Taq PCR Mix: 12.5μl

[0052] ITS4R / 5F primer: 1 / 1μl

[0053] DNA template: 2μl

[0054]The PCR reaction conditions are: 94°C for 5min; 94°C for 1min, 50°C for 1min, 72°C for 1.5min+3s / cycle, 30cycles; 72°C for 7min;

[0055] 4. Sequencing

[0056] The PCR products were dire...

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Abstract

The invention relates to a method for identifying Chinese angelica and its adulterants by utilizing an ITS sequence. The method comprises the following steps: 1, extracting total DNA of the traditional Chinese medicine Chinese angelica and its adulterants; 2, carrying out a PCR amplification reaction by using primers represented by SEQ ID NO.1 and SEQ ID NO.2 respectively with the DNA extracted in step 1 as a template; 3, directly sequencing PCR products; and 4, splicing sequences, contrasting, establishing an NJ tree, and identifying the traditional Chinese medicine Chinese angelica and its adulterants. The ITS sequence is used to identify the traditional Chinese medicine Chinese angelica and its adulterants for the first time. The method has the advantages of simple operation, substantial result, and strong practicality, can realize the rapid and accurate discrimination of Chinese angelica and its adulterants, and is adapted to wide popularization application.

Description

technical field [0001] The invention relates to the technical field of identification of source varieties of Chinese medicinal materials, in particular to a method for identifying angelica sinensis and its counterfeit products by using ITS sequences. Background technique [0002] Angelica sinensis, recorded in the 2010 edition of Pharmacopoeia as the dried root of Angelica sinensis (Oliv.) Diels. For blood deficiency and yellow complexion, dizziness and palpitations, irregular menstruation, amenorrhea dysmenorrhea, abdominal pain due to deficiency and cold, rheumatism arthralgia, injury from falls, carbuncle sores, dryness of the intestine and constipation. Modern pharmacological studies have also shown that Angelica is effective in promoting hematopoiesis and anti-anemia, inhibiting platelet aggregation, anti-thrombosis, anti-coagulation, anti-oxidation, anti-inflammation, anti-tumor, improving the activity of immune factors, neuroprotection and repair, and on the cardiovas...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6809C12Q2531/113C12Q2563/185
Inventor 黄林芳郑司浩林余霖陈士林
Owner INST OF MEDICINAL PLANT DEV CHINESE ACADEMY OF MEDICAL SCI
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