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Mass spectrum guided separation method of cyclopamine analogues in fritillaria plants

A technology for cyclopamine and analogues, which is applied in the field of mass spectrometry-guided separation of cyclopamine analogues in Fritillary plants, can solve the problems of enhancing the orientation and sensitivity of the separation process, and having no solutions

Active Publication Date: 2016-10-26
NANJING FORESTRY UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is still no practical solution on how to enhance the orientation and sensitivity of the separation process

Method used

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  • Mass spectrum guided separation method of cyclopamine analogues in fritillaria plants
  • Mass spectrum guided separation method of cyclopamine analogues in fritillaria plants
  • Mass spectrum guided separation method of cyclopamine analogues in fritillaria plants

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] 1. Experimental part

[0039] 1.1 Reagents and materials

[0040] Reagents: chromatographically pure acetonitrile and chromatographically pure methanol (Merck, Germany), chromatographically pure formic acid (purity 99%, ROC, USA), analytically pure ammonium formate (Shanghai Lingfeng Chemical Reagent Co., Ltd., Shanghai, China). Double distilled water (18MΩ·cm -1 ) made by Milli-Q pure water meter.

[0041] Reference substance: 15 isosteroidal alkaloids, including 5 jervine-type isosteroidal alkaloids: cyclopamine (CA), jervine (JV), peimisine (PS), cycloposine (CS ) and pseudojervine (PJV); 5 cevanine-type isosteroidal alkaloids: puqiedine, puqiedinone, hupehenine, verticine and verticinone; 5 veratramine-type isosteroidal alkaloids (veratramine-type isosteroidal alkaloids): veratrosine (VS), puqienine A, puqienine B, puqienine C, and puqienine D.

[0042] Experimental material: MCX cartridge (Waters Oasis@, mixed strong cation exchange solid phase extraction cartr...

Embodiment 2

[0091] 1. Automatic purification system to separate fritillaria in Fritillaria

[0092] The results of mass spectrometry showed that the isosteroid alkaloid fritillaria m / z 428.4 peaked at 14.0min, and its content was very high. The target compound was separated by an automatic purification system. First establish the total ion chromatogram of fritillaria m / z 428.4 automatic purification system Image 6 (B), according to the extraction chromatogram of the target compound m / z 428.4 see Image 6 (A) Determine the initial collection position, and use the mass spectral response loudness at this position as the mass spectral response intensity threshold (MIT). Finally, it was determined that when the injection volume was 400 μL (concentration about 92 mg / mL), the separation effect was better, and the corresponding response intensity threshold was set to 1600000. At this time, m / z 428.4 peaked at 18.0-19.5 min, and the target compound Collection was performed to obtain fraction m / ...

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Abstract

The invention relates to a mass spectrum guided separation method of cyclopamine analogues in fritillaria plants. According to the method, common product ions of a jerveratrum isosteroidal alkaloid reference substance are taken as the guide, an LC-QTOF-MS / MS technology is introduced to establish a rapid characterization method of jerveratrum isosteroidal alkaloid; then a series of cyclopamine analogues is screened out from fritillaria herbals by using the strong screening functions of LC-QQQ-MS / MS, and finally an automatic purification system is adopted to prepare fractions according to the mass spectrum response signals of fractions.

Description

technical field [0001] The invention relates to a method for separating natural medicines, and discloses a method for separating cyclopamine analogues in Fritillaria plants guided by mass spectrometry. Background technique [0002] Cyclopamine is a kind of veratrum isosteroidal alkaloid, which mainly exists in Liliaceae plants Fritillaria pallidiflora Schrenk, Veratrum californicum and Veratrum grandiflorum. Studies on cyclopamine from the 1960s to the 1980s were limited to teratogenic effects, but studies in the mid-1990s showed that cyclopamine is an inhibitor of the hedgehog signaling pathway, and that mutations in the hedgehog signaling pathway are associated with a variety of tumors Therefore, cyclopamine, as a potential new anti-tumor drug, has set off a research boom all over the world. In recent years, researchers have conducted more in-depth research on cyclopamine, and found that it has good inhibitory activity on medulloblastoma, cholangiocarcinoma, ovarian cance...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/14
CPCG01N30/02G01N30/14G01N2030/027
Inventor 姜艳李会军杜园
Owner NANJING FORESTRY UNIV
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