基于稀疏偏最小二乘回归分析白及抗氧化谱效关系

Spectrum-Effect Relationship Analysis for Antioxidant of Bletilla striata(Thunb.) Reichb. f. Based on Sparse Partial Least Squares Regression

目的:分析白及的抗氧化谱效关系,初步探索白及的抗氧化活性成分。方法:通过D101大孔树脂对白及提取物进行水、30%、50%、70%、95%乙醇分段洗脱,收集12批白及抗氧化效果最好的洗脱段作为试验材料分别建立高效液相(High performance liquid chromatography,HPLC)指纹图谱和进行抗氧化试验,随后通过稀疏偏最小二乘回归法建模进行指纹图谱和抗氧化效果的谱效分析,根据谱效分析结果进行有效峰的二级质谱(MS/MS)初步鉴定。结果:70%乙醇洗脱段的抗氧化效果最好,12批样品1,1-二苯基-2-三硝基苯肼自由基(1,1-diphenyl-2-picrylhydrazyl,DPPH)试验的IC 50范围为(57.883±1.077~135.948±2.096)μg·mL^-1,2,2'-联氮-双-3-乙基苯并噻唑啉-6-磺酸自由基(2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid),ABTS)试验的IC 50为(11.510±0.074~19.180±0.242)μg·mL^-1,其中12批样品的ABTS试验的IC 50均小于维他命E(Vitamin E,VE)。以DPPH为指标时,峰4、12、13、16为主要药效峰,以ABTS为指标时,峰8、13、15为主要药效峰;峰4、8、12、13、15、16经MS/MS初步鉴定为Lusianthridin,3,7-二羟基-2,4-二甲氧基菲,3-[(4-羟苯基)甲基]-4-甲氧基-9,10-二氢菲-2,7-二醇,Bletillatin D,Blestrianol A,3,3'-二羟基-2-(4-羟苄基)-5-甲氧基联苄,其中Bletillatin D与DPPH和ABTS自由基的清除效果均有较好的相关性,是一个潜在的抗氧化剂。结论:经大孔树脂洗脱后,白及的抗氧化效果显著提升,其抗氧化效果可能与其所含化合物Lusianthridin,3,7-二羟基-2,4-二甲氧基菲,3-[(4-羟苯基)甲基]-4-甲氧基-9,10-二氢菲-2,7-二醇,Bletillatin D,Blestrianol A,3,3'-二羟基-2-(4-羟苄基)-5-甲氧基联苄有关。

Objective:The antioxidant components of Bletilla striata were initially explored based on spectrum-effect relationship.Methods:B.striata extract was eluted via D101 macroporous resin with water,30%,50%,70%,95%ethanol in turn.Then,HPLC fingerprint and antioxidant experiments of 12 batches of B.striata were conducted after selecting the best elution fraction.Based on this,spectrum-effect relationship was established by partial least squares regression.Finally,effective peaks were initially identified by MS/MS based on spectrum-effect relationship.Results:70%ethanol elution showed the best antioxidant efficiency.Besides,70%ethanol elution of 12 batches of B.striata showed good antioxidation.All IC 50 values for DPPH assays were in the range of(57.883±1.077-135.948±2.096)μg·mL^-1,and for ABTS assays were(11.510±0.074-19.180±0.242)μg·mL^-1.The IC 50 of all samples for ABTS assays were lower than the IC 50 of VE.In addition,the main antioxidant components for DPPH were peak 4,12,13,16 and for ABTS were peak 8,13,15.And peak 4,8,12,13,15,16 were tentatively identified as Lusianthridin,3,7-dihydroxy-2,4-dimethoxyphenanthrene,3-[(4-hydroxyphenyl)me-thyl]-4-methoxy-9,10-dihydrophenanthrene-2,7-diol,Bletillatin D,Blestrianol A and 3,3'-dihydroxy-2-(4-hydroxybenzyl)-5-methoxybibenzyl,among them,Bletillatin D is a potential antioxidant,which showed a good correlation with the scavenging efficiency of DPPH and ABTS free radicals.Conclusion:The antioxidant efficiency of B.striata was significantly improved by D101 macroporous resin elution.And its antioxidant efficiency might be related to Lusiantridin,3,7-dihydroxy-2,4-dimethoxyphenanthrene,3-[(4-hydroxyphenyl)methyl]-4-methoxy-9,10-dihy-dro-phenanthrene-2,7-diol,Bletillatin D,Blesterinol A,3,3'-dihydroxy-2-(4-hydroxyben-zyl)-5-methoxybiphenyl.