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Method for detecting monosaccharide contents in osmanthus fragrans by pre-column derivatization HPLC-MS/MS

A derivatization and monosaccharide technology, applied in the field of pre-column derivatization HPLC-MS/MS detection of monosaccharide components in sweet-scented osmanthus, to achieve high separation efficiency, good separation, and short analysis time

Inactive Publication Date: 2019-01-11
HUBEI UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the application of PMP pre-column derivatization high performance liquid chromatography coupled with electrospray ionization mass spectrometry in sweet-scented osmanthus has not been reported yet.

Method used

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  • Method for detecting monosaccharide contents in osmanthus fragrans by pre-column derivatization HPLC-MS/MS
  • Method for detecting monosaccharide contents in osmanthus fragrans by pre-column derivatization HPLC-MS/MS
  • Method for detecting monosaccharide contents in osmanthus fragrans by pre-column derivatization HPLC-MS/MS

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] [Example 1] Sample Processing

[0048] a, the extraction of polysaccharides in osmanthus fragrans

[0049] Accurately weigh 10g of sweet-scented osmanthus and 200ml of absolute ethanol into a 500ml round-bottomed flask and mix them, condense and reflux in a constant temperature water bath for one hour at 80°C, take the filter residue after filtration, repeat this operation, filter and dry again; Then the filter residue obtained is placed in a 1000ml round-bottomed flask and put into 400ml of water, extracted in a constant temperature water bath for 1.5h at 90°C, and repeated twice; after suction filtration, the filtrate was obtained, and the filtrates were combined twice, Evaporate and concentrate the filtrate to 150ml in a rotary evaporator; add 4 times the volume of absolute ethanol to the concentrated polysaccharide solution, then centrifuge for 15min (8000r / min) in a centrifuge, and dry to obtain sweet-scented osmanthus crude polysaccharide; add 50mL of distilled wa...

Embodiment 2

[0056] [Example 2] HPLC-MS / MS pre-column derivatization and quantitative analysis

[0057] 1. Instruments and reagents:

[0058] Osmanthus osmanthus, osmanthus osmanthus, laurel osmanthus (Guangxi) (Hubei Jishantang Chinese Herbal Pieces Co., Ltd.), acetonitrile (chromatographically pure), methanol (chromatographically pure), glacial acetic acid (chromatographically pure), absolute ethanol (analytical pure) , acetone (analytical pure), chloroform (analytical pure) (Tianjin Oubokai Chemical Co., Ltd.), n-butanol, ammonia water (analytical pure, Tianjin Hengxing Chemical Reagent Manufacturing Co., Ltd.), hydrochloric acid (analytical pure, Xinyang Chemical Reagent Factory ), ammonium acetate (chromatographically pure, Tianjin Kemiou Chemical Reagent Co., Ltd.), 1-phenyl-3-methyl-5-pyrazolone (PMP), D-mannose (≥99.0% (GC) Sinopharm Chemical Reagent Co., Ltd.), D-glucose, D-xylose, D-galactose (≥99.0% (GC), purchased by Aladdin), D-ribose, L-fucose (≥99.0% (GC) , Cooler Chemical...

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Abstract

The invention discloses a method for detecting monosaccharide components in osmanthus fragrans by pre-column derivatization HPLC-MS / MS. The method uses 1-phenyl-3-methyl-5-pyrazolone (PMP) pre-columnderivatization as a pretreatment mean, and uses high performance liquid chromatography coupled with electrospray ionization mass spectrometry to simultaneously determine 6 kinds of the monosaccharidecomponents in the osmanthus fragrans. The method has the advantages of simple, sensitive, high selectivity, high separation efficiency, small injection amount, short analysis time, multi-mode determination and the like, and provides a scientific basis for the study of the monosaccharide components of polysaccharides in the osmanthus fragrans. Meanwhile, the method lays the groundwork for the studyof other natural products.

Description

technical field [0001] The invention belongs to the field of polysaccharide component analysis, in particular to a method for detecting monosaccharide components in osmanthus fragrans by pre-column derivatization HPLC-MS / MS. Background technique [0002] Osmanthus fragrans Lour, belonging to the family Oleaceae, is an ornamental plant used as a traditional folk medicine in southern and central China for the treatment of various diseases. It has been used as a medicinal plant in China for thousands of years and has a wide range of biological effects, including reducing inflammation, acting as an antioxidant, preventing aging and inhibiting melanin production. In addition, the main bioactive components in Osmanthus fragrans extract are carotenoids and phenolic acids. The analysis of polysaccharides from natural medicines is extensive. The study of polysaccharides as drugs began in the 1950s, and up to now, hundreds of polysaccharides have been found to have biological activi...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06
Inventor 范宝磊李婷婷
Owner HUBEI UNIV OF SCI & TECH
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