Separation And Analysis Of Active Components In Apple Pomace And Green Onion Roots And Neuroprotective Effect On Parkinson's Model Mice

Apples and green onions are common fruit and vegetable,which are rich in resources in our country with huge yields and good economic benefits.However,the accumulation of apple pomace and green onion roots which are by-products of apples and green onions has caused certain environmental problems and waste of useful materials.There are many active ingredients in these by-products.Extracting,separating and purifying these active substances are a vital way to realize the role and value of these by-products.Isoquercitrin is one of the active ingredients in apple pomace,which has good anti-oxidant,anti-inflammatory and other biological activities.Parkinson's disease(PD)is a chronic motor-controlled neurodegenerative disease that seriously affects the normal life of patients.Many studies have shown that its pathogenesis is related to oxidative stress,neuroinflammation,and mitochondrial dysfunction.There is currently no drug that can completely cure PD.Therefore,isoquercitrin separated from apple pomace was administered to Parkinson's model mice induced by MPTP to explore whether it had a neuroprotective effect on these PD model mice and whether it could prevent or delay the PD process.The active ingredients in green onion roots include volatile oils and non-volatile flavonoids.In this paper,GC-MS and GC�GC-TOF/MS were selected to analyze the volatile ingredients in the roots of green onions,and high-speed countercurrent chromatography(HSCCC)was used to separate flavonoids from green onion roots.There are four research results as follows:1.The HPLC analysis method and the HSCCC separation method of isoquercitrin in apple pomace were established.A total of 9.8 g crude extract was obtained from 50 g apple pomace powder.The isoquercitrin with a purity of 96.85%was isolated from apple pomace crude extract by HSCCC.Isoquercitrin was administered to Parkinson's model mice.The behavioral ability of mice was evaluated by rotarod test,pole-climbing test,and suspension test and the results were verified at the molecular level through oxidative stress,immunofluorescence,immunohistochemistry experiments.It was found that isoquercitrin had a neuroprotective effect on MPTP-induced Parkinson's model mice and it was expected to be used in the treatment or improvement of Parkinson's disease.2.The volatile oil in green onions roots was obtained by steam distillation extraction.A total of 126.9 mg volatile oil was obtained from 50.00 g green onion roots powder,and the oil yield was 0.25%.One-dimensional GC-MS separation and identification confirmed 89 compounds,and comprehensive two-dimensional GC�GC-TOF/MS separation and identification confirmed 256 compounds.The separation performance of the comprehensive twodimensional GC�GC-TOF/MS was much higher than that of GC-MS,with higher sensitivity,resolution and peak capacity,which made the analysis and identification of the volatile oil components of green onion roots more accurate.3.The HPLC analysis method of flavonoids in the crude extract of green onion roots was optimized,and a high-speed countercurrent chromatography method was established to separate flavonoids of green onion roots.By comparing different solvent systems,the n-C6H14/CH3COOC2H5/CH3OH/H2O(3:7:5:5,v/v)was found to have the best effect.Therefore,this system was chosen as the solvent system for separating the target substance in the crude extract of green onion roots.A target active substance with a purity of 91.43%was isolated from the crude extract of green onion root,and the other 5 target substances were enriched.