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Quick identification method for ingenol

A fast technology for ingenol, applied in the field of rapid identification of ingenol by high-performance thin-layer chromatography, can solve problems such as complex structure, high detection cost, and long liquid chromatography analysis time

Inactive Publication Date: 2017-12-22
INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the large number of components and complex structure in Capper, different compounds may have the same or similar retention characteristics, which may easily lead to errors in qualitative results
In addition, the analysis time of liquid chromatography is long, the consumption of organic mobile phase is large, and the detection cost is easy to be high, so it is not suitable for the rapid detection and identification of ingenol

Method used

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  • Quick identification method for ingenol
  • Quick identification method for ingenol

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Take 1 mg of ingenol reference substance in a volumetric flask, add 1 mL of methanol to dissolve it completely, and use it as the reference substance solution. In addition, get capers ethanol extract and reclaim the solvent upper layer, adjust the pH value to 12-14, hydrolyze at room temperature for 0.5h, and make the test solution according to [0009]; draw the above-mentioned reference solution and the test solution according to the thin-layer chromatography experiment 5 uL of each product solution was spotted on the same GF254 silica gel plate, developed (the mixture of petroleum ether and ethyl acetate was used as the developing solvent, the volume ratio of petroleum ether:ethyl acetate=3:7), taken out and dried, Spray with 5% vanillin-concentrated sulfuric acid solution (vanillin 5g, 10% sulfuric acid ethanol solution 100 mL)

[0026] For color development, heat until the spots are clear, and observe at 365nm; in the thin-layer chromatography of the reference substa...

Embodiment 2

[0028] Get caperia caperi ethanol extract and reclaim the upper layer of solvent and adjust the pH value to 12-14, room temperature hydrolysis 1h, be made into need testing solution according to [0009]; draw each 5 uL of above-mentioned need testing solution according to thin-layer chromatography experiment, and The reference solution was spotted on the same GF254 silica gel plate, developed (the mixture of petroleum ether and ethyl acetate was used as the developing solvent, the volume ratio of petroleum ether: ethyl acetate = 3:7), taken out and dried, and sprayed with 5% Vanillin-concentrated sulfuric acid solution (vanillin 5g, 10% sulfuric acid ethanol solution 100 mL) develops color, heats until the spots are clear, and observes at 365nm; Light blue, other spots appear orange, cyan or pink, see details figure 1 Point 3 in .

Embodiment 3

[0030] Get capers ethanol extract and reclaim solvent upper layer to adjust pH value to 12-14, room temperature hydrolysis 2h, be made into need testing solution according to [0009]; Draw each 5 uL of above-mentioned need testing solution according to thin-layer chromatography experiment, and The reference solution was spotted on the same GF254 silica gel plate, developed (the mixture of petroleum ether and ethyl acetate was used as the developing solvent, the volume ratio of petroleum ether: ethyl acetate = 3:7), taken out and dried, and sprayed with 5% Vanillin-concentrated sulfuric acid solution (vanillin 5g, 10% sulfuric acid ethanol solution 100 mL) develops color, heats until the spots are clear, and observes at 365nm; Light blue, other spots appear orange, cyan or pink, see details figure 1 Point 4 in .

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Abstract

The invention belongs to the technical field of natural medicinal chemistry and particularly relates to a quick identification method for ingenol. The method successively includes the steps of: 1) performing alkali hydrolysis, neutralization and extraction to an upper layer obtained after solvent recovery to an alcohol extract liquid of Euphorbia lathyris, volatilizing the solvent out, and adding methanol for dissolution; 2) collecting a supernatant and performing thin-layer chromatographic experiment, wherein the sample is developed with petroleum ether (60-90 DEG C) - ethyl acetate as a developing agent; 3) taking a thin-layer plate out, air-drying the thin-layer plate, spraying a color developing agent onto the thin-layer plate for color development, and heating the thin-layer plate until stripes can be clearly seen; 4) observing the thin-layer plate with wavelength being 365 nm, wherein in a spectral map, light-blue color spots represent for the ingenol, while orange, cyan and pink spots represent for other substances. The method is high-effective, simple, accurate and reliable, fills the gap of specific quick detection and identification for the ingenol, and can be used for quick detection of the ingenol.

Description

technical field [0001] The invention belongs to the technical field of chemical identification of natural medicines, and in particular relates to a rapid identification method of ingenol by high-efficiency thin-layer chromatography. Background technique [0002] Ingenol was first obtained from Euphorbiaceae in 1968 E It is isolated from ingens and has a unique high-tension 5 / 7 / 7 / 3 tetracyclic core. It interacts with protein kinase C to produce pharmacological activities such as anti-cancer and anti-HIV. [0003] In 2012, Ingenol mebutate (drug name Picato) developed by LEO Pharmaceuticals was approved by the US FDA and became the first drug for the treatment of actinic keratosis. Gel formulation. In addition, ingenyl methacrylate can also be used in the treatment of basal cell carcinoma, and phase II clinical trials are underway. The production of ingenol methyl crotonate is obtained by semi-synthesis with ingenol as raw material, and the yield rate is 62%. However, inge...

Claims

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Application Information

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IPC IPC(8): G01N30/90
CPCG01N30/90
Inventor 姜杉陈雨赵兴增刘飞吕慧王鸣冯煦
Owner INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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