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Construction method of HPLC fingerprint and DNA fingerprint of Saussurea Involucrata

A technology of DNA fingerprinting and fingerprinting, which is applied in biochemical equipment and methods, microbiological measurement/inspection, material separation, etc., to achieve the effect of large amount of characterization information, good repeatability and simple method

Inactive Publication Date: 2009-04-29
INST OF PROCESS ENG CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] There is no report on DNA fingerprinting of Snow Lotus

Method used

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  • Construction method of HPLC fingerprint and DNA fingerprint of Saussurea Involucrata
  • Construction method of HPLC fingerprint and DNA fingerprint of Saussurea Involucrata
  • Construction method of HPLC fingerprint and DNA fingerprint of Saussurea Involucrata

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Establishment of Xinjiang Snow Lotus HPLC Fingerprint

[0032] 1. Instruments and materials

[0033] Shimadzu LC-20A chromatograph, equipped with autosampler, PDA diode array detector, LC solution chromatographic workstation, Shimadzu Corporation, Japan; LTQ liquid mass spectrometer, Thermoelectric Company of the United States; organic phase filter membrane d: 50mm, pore size: 0.22μm, organic phase syringe filter 13mm×0.20μm, Shanghai Anpu Company; 40-mesh standard sieve, aperture: 0.45μm: Shangyu Instrument Factory; JY99-2D ultrasonic cell pulverizer; Ningbo Xinzhike Instrument Research Institute; TDL-5-A low-speed desktop centrifuge, Shanghai Anting Scientific Instrument Factory; ultrapure water device, Millipore, USA; RE-52AA rotary evaporator, Shanghai Yarong Biochemical Instrument Factory; KQ5200DB CNC ultrasonic cleaner, Yuhuan, China dawn peak.

[0034] Methanol, acetonitrile (chromatographic alcohol J&K Chemical); anhydrous methanol, acetic acid (ana...

Embodiment 2

[0063] Example 2 The establishment method of the Xinjiang Snow Lotus DNA fingerprint:

[0064] 1. Extraction of DNA

[0065] Grinding samples of dried leaves of medicinal materials from various origins were taken, and their DNA was extracted by CTAB method. The purity and concentration of the obtained DNA were identified by gel electrophoresis combined with density scanning method.

[0066] 2. PCR amplification

[0067] The PCR reaction was carried out on a PCR amplification instrument modeled as Longene MG-96G, and the reaction system was 25ul. in,

[0068] RAPD analysis includes: template genome 50ng DNA, 2.3mmol / LMg 2+ , 1.8UTaq enzyme, 0.35mmol / LdNTP, 0.6umol / L primer, the basic amplification program is: 95°C pre-denaturation for 2min, 94°C denaturation for 40s, 37°C annealing for 1min, 72°C extension for 2min, 40 cycles, and finally 72°C Extend for 7min.

[0069] ISSR analysis includes: template genome 50ng DNA, 2.5mmol / L Mg 2+ , 2.5U Taq enzyme, 0.35mmol / LdNTP, 0.8...

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Abstract

The invention relates to an HPLC fingerprint spectrum of Saussurea involucrate and a method for establishing a DNA fingerprint spectrum of the Saussurea involucrate. The invention is characterized in that the HPLC fingerprint spectrum comprises fingerprint spectrums of two extraction positions with different polarities of ethyl acetate and n-butyl alcohol, and the fingerprint spectrum of the extraction position of the ethyl acetate has 20 common peaks, wherein 7 peaks refer to scopoletin, rutoside, isoquercitrin, xylostein, quercetrin, homoplantaginin and hispidulin respectively; the fingerprint spectrum of the extraction position of the n-butyl alcohol has 11 common peaks, wherein 3 peaks refer to bergapten, scopoletin and rutoside; and the DNA fingerprint spectrum consists of an RAPD molecular fingerprint spectrum and an ISSR molecular fingerprint spectrum. The quality and the reality and falsity of the Saussurea involucrate can be universally and effectively detected in the aspects of chemistry and biology by combination of HPLC fingerprint and DNA fingerprint.

Description

technical field [0001] The invention relates to a method for establishing an HPLC fingerprint and a DNA fingerprint of Xinjiang snow lotus, which can detect the quality of Xinjiang snow lotus from two angles of chemistry and biology. Background technique [0002] Xinjiang snow lotus whole herb is used as medicine, which is the mainstream variety of snow lotus medicinal materials. Snow lotus has a long history of medicinal use. It is sweet, bitter, and warm, and belongs to the liver, spleen, and kidney meridians. It has the effects of clearing away heat and detoxification, expelling wind and dampness, promoting menstruation and promoting blood circulation, strengthening muscles and yang, anti-inflammatory and analgesic, warming the palace and dispelling cold; it is used for rheumatoid arthritis, cold palace abdominal pain, amenorrhea, retained placenta, stroke, kidney deficiency low back pain, Nocturnal emission, impotence, measles impotence, lung-cold cough, high mountain i...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N30/02
Inventor 袁晓凡赵兵王晓东王美玲戴卓昊欧元
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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