Rapid analysis method of unknown sesquiterpenes pyridine alkaloids in thunder god vine

A rapid analysis technology for semiterpene pyridine, which is applied in the field of medicine, can solve problems such as difficulties in natural products, limited database, poor reproducibility of secondary mass spectrometry, etc., and achieve the effect of improving reliability and avoiding cumbersome data

Inactive Publication Date: 2015-08-19
CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

At present, the main analysis is NMR, but NMR identification needs to obtain pure products, so it is impossible to quickly analyze the relatively low content of unknown sesquiterpene pyridine alkaloids in Tripterygium wilfordii
Liquid-mass spectrometry is an important method for analyzing unknown trace substances in complex systems. Current methods often require databases (J.Am.Chem.Soc.2009, 131, 5030; Anal.Chem.2008, 80, 1280), but the database of LC-MS is very limited, and the reproducibility of MS / MS between different instruments is poor. In addition, it is very difficult to obtain trace amounts of standard natural products; there are many isomers in plants Constructs increase the difficulty of analysis

Method used

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  • Rapid analysis method of unknown sesquiterpenes pyridine alkaloids in thunder god vine
  • Rapid analysis method of unknown sesquiterpenes pyridine alkaloids in thunder god vine
  • Rapid analysis method of unknown sesquiterpenes pyridine alkaloids in thunder god vine

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Embodiment 1

[0024] (1). Extraction of Active Components of Tripterygium wilfordii

[0025] Tripterygium wilfordii was purchased from the Hehuachi market in Chengdu, soaked in 100% methanol ultrasonically for one hour, then concentrated and filtered under low pressure to obtain the corresponding solid.

[0026] (2). Analysis of fragment ions of known single sesquiterpene pyridine alkaloids

[0027] Instrument: Bruker ESI-Q-TOF (micrOTOF Q)

[0028] Mass spectrometry conditions: ionization mode, positive ion mode (+); spray gas pressure, 30psi; argon as collision gas, collision energy, 25eV; drying gas temperature, 180°C; capillary port voltage, -4500V, metal cover voltage, - 4000V.

[0029] The secondary spectrum of the known compound cangorinine E-1 is analyzed (attached Figure 1 (a)), the study found that the fragment ions in the high-mass region are mainly lost by breaking the side chain, and the fragment ions in the low-mass region, such as m / z 206, are mainly the characteristic ion...

Embodiment 2

[0038] (1). Extraction of Active Components of Tripterygium wilfordii

[0039] Tripterygium wilfordii was purchased from the Hehuachi market in Chengdu, soaked in 100% methanol ultrasonically for one hour, then concentrated and filtered under low pressure to obtain the corresponding solid.

[0040] (2). Analysis of fragment ions of known single sesquiterpene pyridine alkaloids

[0041] Instrument: Bruker ESI-Q-TOF (micrOTOF Q)

[0042]Mass spectrometry conditions: ionization mode, positive ion mode (+); spray gas pressure, 30psi; argon as collision gas, collision energy, 25eV; drying gas temperature, 180°C; capillary port voltage, -4500V, metal cover voltage, - 4000V.

[0043] The secondary spectrum of the known compound euonymine is analyzed (attached Figure II (a)), the study found that the fragment ions in the high-mass region are mainly lost by breaking side chains, and the fragment ions in the low-mass region are mainly characteristic ions of the pyridine part.

[00...

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Abstract

The invention relates to a rapid mass spectrometry method for indentifying unknown sesquiterpenes pyridine alkaloids in thunder god vine, which belongs to the pharmacy field. The method comprises the following steps: processing Chinese fevervine herb by a pre-treatment process of a sample through solvent ultrasonic extraction; using an electro-spray, a quadrupole and a time-of-flight mass spectrometer for analyzing fragment ions of the known single sesquiterpenes pyridine alkaloid, especially analyzing the fragment ions through frame cracking; using a high performance liquid chromatography, the electro-spray and the quadrupole and the time-of-flight mass spectrometer to obtain a secondary mass spectrum of sesquiterpenes pyridine alkaloids in a complex system to obtain the fragment ions of an unknown compound, wherein a profile diagram of the fragment ions of sesquiterpenes pyridine alkaloid in the complex system can be respectively obtained by using two methods; comparing the profile diagram of the fragment ions of the sesquiterpenes pyridine alkaloid in the known complex system and the complex system and comparing the characteristic fragment ions, and performing a rapid structural analysis on the unknown sesquiterpenes pyridine alkaloid in the complex system.

Description

technical field [0001] The invention belongs to the technical field of medicine, and relates to independent and rapid analysis of unknown sesquiterpene pyridine alkaloid components in Tripterygium wilfordii. Background technique [0002] Tripterygium wilfordii Hook.f. is a plant of the genus Tripterygium wilfordii Hook.f., which is mainly distributed in southern China and is an important traditional Chinese medicinal material. It is used to treat autoimmune diseases such as rheumatoid arthritis, chronic nephritis, lupus erythematosus, and contact dermatitis, all of which have significant curative effects. [0003] Sesquiterpene pyridine alkaloids are a very important active ingredient in Tripterygium genus. Studies have shown that this type of compound has anti-immune, insecticidal, anti-HIV (J.Nat.Prod.2000,63,357), anti-cancer, etc. active. Therefore, it has attracted many scientists to conduct research. Efficient analytical methods are crucial to research in the biolog...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88
Inventor 吴志军方冬梅蒋艳戚华溢张国林
Owner CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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