【作者】 张莉；

【导师】 郑磊；

【作者基本信息】 合肥工业大学 ， 食品工程（专业学位）， 2021， 硕士

【摘要】 多花黄精(Polygonatum cyrtonema Hua)为百合科(Liliaceae)黄精属(Polygonatum Mill.)植物,是一种常见的药食同源植物,可经黄酒“九蒸九制”炮制。目前,对于多花黄精炮制前后药理活性和成分的差异性比较,多集中于醇提物成分,而缺乏水提物相关活性成分的比较研究。因此,本论文以“炮制前后的多花黄精水提物”(以下简称为“炮制前后水提物”)为切入点,比较了其抗氧化活性与相关成分的变化。本论文,首先将炮制前后的多花黄精粉末,经热水浸提获得炮制前后的水提物,比较其体外抗氧化活性,发现“九蒸九制”后水提物总抗氧化能力(Total antioxidant capacity,T-AOC)和2,2-Diphenyl-1-picrylhydrazyl(DPPH)自由基清除能力,均得到显著性提高。此后,将炮制前后的水提物,经醇沉,取上清液再经离心、浓缩、干燥以获得到脂溶性组分,取醇沉物再经除蛋白、透析、干燥以得到多糖组分。通过测定T-AOC和DPPH等指标,发现炮制后水提物脂溶性组分和多糖组分抗氧化活性,均显著高于炮制前。对于脂溶性组分,利用优化后的HPLC分析条件:色谱柱ZORBAX SB-C18(5μm,4.6×250 mm)、进样20μL、波长203 nm、柱温30℃、流速1.0 m L/min;流动相水和乙腈梯度洗脱;再经HPLC-ESI-MS/MS技术,鉴别了20种化合物。Guanosine和L-Tryptophan炮制后含量显著降低,Adenosine、Cyclo dileucyl(or isoleucyl)、Polygonatine B、4-(1-methyl-dodecyl)-benzene Sulfonic acid和Linoleic acid在炮制后含量显著升高。对于多糖组分,通过HPGPC法对炮制前后的分子量进行探究,发现炮制前多糖(PCPC)为均一对称峰,分子量4842 Da;炮制后多糖(PCPW)为三个重叠峰,分子量分为1.69×10~7Da、7.96×10~6Da和3.39×10~6Da。单糖组成分析显示,PCPC主要为果糖(93.57%),另含有葡萄糖(6.43%);PCPW主要也是果糖(95.35%),另含有则葡萄糖(3.00%)、鼠李糖(0.50%)、半乳糖(0.05%)、甘露糖(0.83%)、半乳糖醛酸(0.15%)和葡萄糖醛酸(0.14%)。进一步,DEAE cellulose-52色谱柱分析表明,PCPC主要含由蒸馏水洗脱的中性多糖PCPC-1;PCPW既含由蒸馏水洗脱的中性多糖PCPW-1,也含有不同浓度氯化钠溶液洗脱的酸性多糖PCPW-2～PCPW-7。总之,本论文发现,炮制后水提物抗氧化活性显著提升。这一提升,可能与炮制前后脂溶性组分和多糖组分成分的显著变化相关。更多还原

【Abstract】 Polygonatum cyrtonema Hua,a common medicinal and edible homologous plant,belongs to the genus Polygonatum Mill.(Liliaceae),which can be processed by nine steaming and nine processing processes with rice wine.At present,the comparison of pharmacological activity and composition difference of crude and wine-processed Polygonatum cyrtonema Hua is mainly focused on the composition of alcohol extract,but the comparative study of active composition related to aqueous extract is lacking.Therefore,in this paper,the changes of the antioxidant activity and related composition of crude and wine-processed P.cyrtonema Hua aqueous extract(hereinafter referred to as the aqueous extract before and after processing)were compared.In this paper,firstly,the aqueous extract of crude and wine-processed P.cyrtonema Hua was obtained by hot water extraction,and its antioxidant activity in vitro was compared.The total antioxidant capacity(T-AOC)and free radical scavenging ability of2,2-diphenyl-1-picrylhydrazyl(DPPH)of the aqueous extract were significantly increased after nine steam and nine processing.After that,the aqueous extract was precipitated by alcohol,and the supernatant was centrifuged,concentrated and dried to obtain the fat-soluble part.The alcohol precipitate was then deproteinized,dialysed and dried to obtain the polysaccharide part.By measuring the indexes of T-AOC and DPPH,it was found that the antioxidant activity of the fat-soluble and polysaccharide parts of wine-processed P.cyrtonema Hua aqueous extract were significantly higher than those of crude P.cyrtonema Hua aqueous extract.For fat-soluble part,the optimized HPLC analysis conditions were used.Chromatographic column was ZORBAX SB-C18(5μm,4.6×250 mm),injection was20μL,wavelength was 203 nm,column temperature was 30℃,flow rate was 1.0m L/min and gradient elution of mobile phase water and acetonitrile.Then,20 known compounds were found by HPLC-ESI-MS/MS.Among the known compounds,the content of Guanosine and L-tryptophan were significantly reduced while the content of Adenosine、Cyclo dileucyl(or isoleucyl)、Polygonatine B、4-(1-methyl-dodecyl)-benzene sulfonic acid and Linoleic acid were significantly improved after processing.For the polysaccharide part,HPGPC method was used to investigate the molecular weight of the polysaccharide before and after processing.It was found that the polysaccharide before processing(PCPC)had a uniform symmetrical peak with a molecular weight of 4842 Da.After processing,the polysaccharide(PCPW)had three overlapping peaks,and the molecular weights were 1.69×10~7Da,7.96×10~6Da and3.39×10~6.The analysis of monosaccharide composition showed that PCPC was mainly fructose(93.57%)and glucose(6.43%).PCPW was mainly fructose(95.35%),and it also contained glucose(3.00%),rhamnose(0.50%),galactose(0.05%),mannose(0.83%),galacturonic acid(0.15%)and glucuronic acid(0.14%).Further analysis on DEAE Cellulose-52 column showed that PCPC-1,a neutral polysaccharide eluted by distilled water,was mainly contained in PCPC.PCPW contained neutral polysaccharide PCPW-1 eluted by distilled water and acidic polysaccharide PCPW-2～PCPW-7 eluted by sodium chloride solution of different concentrations.In conclusion,the present study found that the antioxidant activity of P.cyrtonema Hua aqueous extract significantly increased after processing,which maight be related to the significant changes in the composition of fat-soluble part and polysaccharide part in P.cyrtonema Hua aqueous extract.更多还原