| Object:The optimal water extraction process of PNP was screened,and the effects of activated carbon decolorization and resin decolorization on the decolorization effect,physical properties and in vitro antioxidant activity of PNP were investigated.Draft PNP quality standards.Methods:1.Taking the extraction rate of PNP as the evaluation index,the effects of solid-liquid ratio,extraction time,extraction temperature and extraction times on the extraction rate were investigated by single factor experiment.On the basis of single factor experiment,four factors and three levels orthogonal experiment were used to optimize the water extraction conditions of PNP.2.With comprehensive score as evaluation index,the optimal decolorization specifications of activated carbon were screened from granular activated carbon and powdered activated carbon,and then the influence of activated carbon dosage,decolorization temperature and decolorization time on the comprehensive score was investigated by single factor experiment,and the decolorization conditions were optimized by three-factor and three-level orthogonal experiment on the basis of single factor experiment.Through static adsorption experiment,the best decolorizing resin was selected from six kinds of resins,and then the influence of sample loading amount,decolorizing time and rotational speed on comprehensive score was investigated by single factor experiment.On the basis of single factor experiment,the decolorizing conditions were optimized by three factors and three levels orthogonal experiment.3.The solubility,particle size,DSC,FT-IR,p H,relative molecular weight and Monosaccharide composition of UPNP,PNPC and PNPD were determined respectively.The scavenging rate of DPPH,ABTS~+and hydroxyl group free radicals,metal ion chelating ability and reducing power of UPNP,PNPC and PNPD were determined respectively.4.The identification method of PNP PMP-derived high performance liquid chromatography for monosaccharide composition was established,and the methods of PNP moisture detection,ash detection,heavy metal detection,relative molecular weight and distribution detection were established.The content of neutral and acidic sugars of PNP was determined by anthrone sulfate method combined with m-hydroxy-biphenyl method.Results:1.According to the orthogonal experiment results of PNP extraction process,the influencing sequence of each experimental factor on the extraction rate of PNP is as follows:C(extraction temperature)>D(extraction times)>B(extraction time)>A(solid-liquid ratio),the optimal extraction process of PNP was C3D2B3A1,that is,extraction temperature 92 ~oC,extraction times 2,extraction time 6 h,solid-liquid ratio1:10,3 batches of verification,the optimal extraction rate of PNP was 11.18%±0.41%.2.The decolorization effect of powdered activated carbon is the best.Orthogonal experiment results showed that the influencing order of each factor on the comprehensive score was A(amount of activated carbon)>B(decolorization time)>C(decolorization temperature),and the optimal decolorization condition is A2B3C1,that is,PNP extract 40 m L,amount of activated carbon 2.0%,decolorization time 45 min.The decolorization temperature was 50℃,and the three tests showed that the retention rate of acid polysaccharide was 91.20%±0.96%,the total sugar retention rate was77.60%±1.40%,the decolorization rate was 65.40%±0.07%,and the comprehensive score was 78.00%.Through static adsorption experiment,D285 resin was selected as the best decolorizing resin from six kinds of resins.Orthogonal experiment results showed that the influencing order of each influencing factor on the comprehensive score was A(sample amount of PNP extract)>B(decolorizing time)>C(rotational speed),and the optimal decolorizing condition was A1B1C3,that is,4 g D285 resin.The sample size of PNP extract was 40 m L,the decolorization time was 1 h,the oscillator speed was 180 r/min,and the three batches were verified:the acid polysaccharide retention rate was 56.50%±1.92%,the total sugar retention rate was 69.70%±0.95%,the decolorization rate was 87.20%±0.34%,the comprehensive score was 71.00%.3.PNP color obtained by no decolorization and two decolorization methods:UPNP>PNPC>PNPD.According to the results of equilibrium solubility experiment,the solubility of UPNP and PNPC had no significant difference(P>0.05),while the solubility of PNPD decreased(P<0.01).According to the particle size analysis results,the PDI of UPNP,PNPC and PNPD are 0.505,0.212 and 0.499,respectively.There are two groups of peaks on the UPNP particle size distribution diagram,corresponding to the average particle size is 228.94 nm and 12230.36 nm,respectively,and the particle size is concentrated around 228.94 nm.PNPC also has two groups of peaks in particle size distribution map,whose corresponding average particle size is 380.87 nm and12420.35 nm,and particle size is concentrated around 380.87 nm.There are three groups of peaks in the particle size distribution diagram of PNPD,corresponding to the average particle size of 44.44 nm,881.44 nm,11729.60 nm,and the particle size is concentrated in the vicinity of 881.44 nm.Only PNPD is distributed in the particle size of less than 100 nm,so it is speculated that PNPD has the break of glycosidic bond in the decolorization process.To produce polysaccharides with smaller molecular weights.According to DSC analysis,UPNP,PNPC and PNPD all have a set of melting peaks near 292℃,ΔH values are 111.1 J/g,175.00 J/g and 145.70 J/g,respectively.The enthalpy values of PNPC are the highest,indicating that PNPC has the highest thermal stability.In addition,there is a set of melting peaks at 157.78℃,ΔH is 86.46 J/g,which is speculated to be caused by two different crystal states in the crystallization process of PNPD.According to FT-IR results,UPNP,PNPC,PNPD and precipitate in PNPD all had characteristic polysaccharide absorption peaks.According to the determination of p H value of PNP extract before and after decolorization,it was found that the p H of PNP extract after decolorization by D285 resin was significantly increased(P<0.01).According to the results of molecular weight determination,the molecular weight of UPNP was 18206 Da and the retention time was 17.70 min,the molecular weight of PNPC was 17644 Da and the retention time was 17.67 min,and the molecular weight of PNPD was 24355 Da and the retention time was 18.82 min.According to the results of monosaccharide composition,UPNP,PNPD and PNPC were all composed of glucose,galactose,arabinose,galacturonic acid,mannose,ribose,rhamnose,glucuronic acid,xylose and fucose.According to the results of in vitro free radical scavenging experiments,the scavenging ability of DPPH and ABTS~+free radicals,the chelating ability of metal ions and the reducing power of PNPC group were significantly higher than those of UPNP group and PNPD group(P<0.01).and the scavenging ability of hydroxyl group free radicals of PNPC group and PNPD group was not significantly different.The scavenging ability of PNPD group on DPPH and ABTS~+free radical was significantly lower than that of UPNP group(P<0.01).4.The identification method of PNP PMP-derived high performance liquid chromatography for monosaccharide composition was established,and the methods of PNP moisture detection,ash detection,heavy metal detection,relative molecular weight and distribution detection were established.The content of neutral and acidic sugars of PNP was determined by anthrone sulfate method combined with m-hydroxy-biphenyl method.Conclusion(s):The optimal water extraction process of PNP was:solid-liquid ratio 1:10,extraction temperature 92 ~oC,extraction times 2,extraction time 6 h.The optimal d ecolorization process of activated carbon was as follows:activated carbon dosage 2.0%,decolorization time 45 min,decolorization temperature 50℃,comprehensive score 78.00%;The optimal decolorization process of D285 resin was 4 g D285 resin,the sam ple size of PNP extract was 40 m L,the decolorization time was 1 h,and the oscillator speed was 180 r/min.The comprehensive score is 71.00%.In the decolorization proce ss of D285 resin,OH~-was dissociated,resulting in a strong local alkaline environment.It is estimated that the PNP glycosidic bond was hydrolyzed,resulting in the distributi on of polysaccharide with particle size less than 100 nm,as well as polysaccharide wit h larger particle size,which resulted in decreased solubility and stability of polysaccha ride and decreased antioxidant activity in vitro.Therefore,activated carbon method wa s the best PNP decolorization method.PNP pilot production process is as follows:750g of pantoginseng residue was extracted,the ratio of material to liquid was 1:10,extra ction temperature was 92 ~oC,extraction time was 6 h,filtration was combined with filt rate,concentration was reduced at 80℃to 5000 m L,centrifugation was performed at8000 r/min for 5 min,supernatant was collected,and 2.0%activated carbon powder w as added,decolorization temperature was 50℃,decolorization time was 45 min.Rem ove the activated carbon,reduce pressure at 80℃and concentrate to 1000 m L,add an hydrous ethanol with triploid volume,overnight at 4℃,filter,take precipitation,and d ry at 80℃to obtain.The quality standard of PNP was established around the identific ation,inspection and content determination of polysaccharide.The quality standard sy stem was simple,stable and feasible. |
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