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Analysis method of characteristic alkaloid composition in fresh Dendrobium huoshanense

A technology of alkaloids and rice dendrobium, which is applied in the field of component analysis of Chinese medicinal materials, can solve the problems of inapplicability and limited reference, and achieve the effect of ensuring separation loss and improving accuracy

Pending Publication Date: 2019-01-29
HEFEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The extraction, separation and detection methods reported above are useful for the analysis of the alkaloid composition of Dendrobium nobile or Dendrobium officinale of the same species, but are not applicable or have very limited reference value for the study of the characteristic alkaloid composition in Huoshan.

Method used

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  • Analysis method of characteristic alkaloid composition in fresh Dendrobium huoshanense
  • Analysis method of characteristic alkaloid composition in fresh Dendrobium huoshanense
  • Analysis method of characteristic alkaloid composition in fresh Dendrobium huoshanense

Examples

Experimental program
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Embodiment 1

[0031] Preparation of the crude extract of Huoshan officinalis

[0032] 300 g fresh strips of Dendrobium Huoshanense, cut into 1 cm segments, freeze-dried and crushed to obtain 50 g of freeze-dried powder.

[0033] Weigh 2g of Huoshan Dendrobii freeze-dried powder, add extraction solvent with a solid-liquid ratio of 1:25, the solvent is 70% ethanol (pH3) or 95% ethanol or chloroform (after ammonia water infiltration), extract three times at 80 ° C, each time 2 hours, the combined extracts were concentrated under reduced pressure to about 30mL to obtain the crude extract of Huoshan rice dendrobium.

[0034] The content of total alkaloids in the crude extract was determined by bromocresol green acid dye-UV-Vis spectrophotometry. The total alkaloid content is calculated as milligrams of total alkaloids dissolved in 1 g of freeze-dried powder, expressed in (‰). The results are shown in Table 1.

[0035] Table 1. Comparison of the total alkaloid content of Dendrobium in different...

Embodiment 2

[0039] Preparation and Composition Analysis of Crude Alkaloids from Huoshan officinalis

[0040] The Huoshan Dendrobii crude extract in Example 1 was concentrated under reduced pressure, added 2% HCl aqueous solution to adjust the pH to 2, extracted three times with dichloromethane (same volume as the acidic water layer) to remove fat; then added ammonia water to the aqueous layer to adjust the pH 10. Extract three times with chloroform (same volume as the alkaline water layer), combine the chloroform, and wash with water until neutral; concentrate under reduced pressure to obtain the crude extract of Huoshanense officinalis alkaloids, dissolve with methanol, and perform HPLC detection respectively ( attached figure 1 ), GC-MS (attached figure 2 ) and LC-MS analysis (attached image 3 ).

[0041] Agilent 1260HPLC analysis conditions are: mobile phase: acetonitrile-0.1% triethylamine solution = 50:50 (V / V); chromatographic column: SGE protocol C18 (5μm, 4.6×250mm); detectio...

Embodiment 3

[0048] Separation and Composition Analysis of Crude Extract of Alkaloids from Orchid Dendrobii by Solid-Phase Extraction Column

[0049] Plexa PCX (60mg / 3cc) solid-phase extraction column was activated with 5mL of methanol and equilibrated with 5mL of pure water, and the crude alkaloids of Huoshanense officinalis were dissolved in 2% H3PO4 solution. Rinse with formic acid aqueous solution, and then elute with 1:1 methanol / acetonitrile solution and 1:1 methanol / acetonitrile solution containing 3%, 5%, and 7% NH3·H2O respectively in 8-10mL (among them, for 1: 1 Methanol / acetonitrile eluent was carried out liquid mass analysis and found that there was no substance), the eluent containing ammonia was concentrated and desalted respectively to obtain alkaloid components I, II, and III, and the liquid mass spectrometry analysis was carried out again, and the analysis results were listed in In Table 2, some substances with undetermined structures, such as: 274.2740, 256.2414 and 246.2...

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Abstract

The invention discloses an analysis method of characteristic alkaloid composition in fresh Dendrobium huoshanense. The method comprises the following steps: taking acid ethanol crude extraction solution of fresh Dendrobium huoshanense strip as a starting raw material, acquiring alkaloid crude extract after performing acid-alkali adjusting-liquid-liquid extraction; initially analyzing possible compositions of the total alkaloids of the Dendrobium huoshanense by combining liquid chromatogram, gas chromatography-mass spectrometry, and liquid chromatography-mass spectrometry; and then separating by adopting a solid-phase extraction column to acquire characteristic alkaloid components, and confirming the alkaloid compositions by means of the liquid chromatography-mass spectrometry technology. The alkaloid compositions in the fresh Dendrobium huoshanense are analyzed and detection, and 11 types of alkaloids with known structures are totally identified, and three types of sesquiterpenoids alkaloids with unknown structures are identified, so that a method for researching the characteristic alkaloids substance of the Dendrobium huoshanense is provided, and the theoretical basis is laid forthe composition-activity relation research.

Description

technical field [0001] The invention relates to the technical field of component analysis of traditional Chinese medicinal materials, in particular to an analysis method for characteristic alkaloid composition in fresh Huoshan rice dendrobium. Background technique [0002] Dendrobium has the effects of nourishing the stomach and promoting body fluid, nourishing yin and clearing heat, etc., which are recorded in the 2010 and 2015 editions of "Chinese Pharmacopoeia". Modern pharmacological studies have shown that Dendrobium has multiple functions such as enhancing immunity, anti-fatigue, anti-oxidation, and preventing radiation damage. Its main active components are polysaccharides, alkaloids, and volatile components. Since ancient times, Dendrobium has paid attention to "fresh products first". Famous prescriptions such as "San Xian Decoction" and "Ye's Nourishing Stomach Decoction" are all made of fresh Dendrobium. Professor Hao Jinda, an expert in modern fresh medicine rese...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06G01N2030/062
Inventor 惠爱玲陈婧超徐涛张文成吴泽宇
Owner HEFEI UNIV OF TECH
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