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
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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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