UV-Vis结合HPLC FP对不同采收期傣药灯台叶的鉴别及品质研究  

Distinguish and Quality Estimation of the Leaves of Alstonia scholaris (L.) R.Br.from Different Harvest Time Based on the UV-Vis·FP and HPLC·FP

作　　者：杨妮娜[1,2] 张霁[3] 赵艳丽[3] 王元忠[3] 赵应红[1] 

机构地区：[1]西双版纳傣族自治州傣医医院,云南西双版纳666100 [2]云南中医学院中药学院,云南昆明650500 [3]云南省农业科学院药用植物研究所,云南昆明650200

出　　处：《光谱学与光谱分析》2016年第12期4021-4027,共7页Spectroscopy and Spectral Analysis

基　　金：国家自然科学基金项目(81260608);云南省自然科学基金重大项目(2013FC006)资助

摘　　要：建立不同采收期灯台叶紫外-可见光谱指纹图谱及HPLC指纹图谱,结合主成分分析、聚类分析对不同采收期灯台叶进行快速鉴别和品质评价,确定最佳采收期,推动傣药现代化发展进程。通过单因素实验确定灯台叶紫外-可见光谱的最佳提取条件,采集12个月份灯台叶紫外光谱数据,平行3次,扣除背景8点平滑后倒入SIMCA-P+11.5进行主成分分析,以前三个主成分三维得分图快速鉴别不同采收期。Agilent ZORBAX Eclipse XDB C18(250×4.6mm,5μm)色谱柱,以乙腈(B)-0.1%甲酸水(A)为流动相,梯度洗脱(0~5min,5%B;5~35min,5%B→26%B;35~40min,26%B→56%B),流速1mL·min^(-1),进样量7μL,柱温30℃,检测波长287nm。不同采收期灯台叶紫外-可见光谱根据吸收峰位置及变化的幅度可以将光谱分为三段,第一段为235~400nm,第二段为400~500nm,第三段为500~800nm。第一段中吸收峰数目最多,主要集中在270,287和325nm,吸光度及其变化幅度最大,体现出不同月份光谱图的指纹特征。第二段吸收峰较少主要分布在410nm和464nm附近,吸光度及其变化较第一段减小。第三段图谱在665nm处均有一个较大吸收峰,吸光度无明显差异。将UV-Vis光谱数据进行主成分分析,不同月份样品在主成分得分图中离散分布,同一月份样品相对集中,可以将不同月份样品鉴别开。HPLC指纹图谱结合聚类分析可将不同采收期样品分为Ⅲ类,第Ⅰ类为3,4,5和7月份,第Ⅱ类为6,8和9月份,第Ⅲ类为10,11,12,1及2月。结合共有峰面积可以看出同一类样品化学成分含量相近,不同类之间差异较明显,第Ⅲ类样品化学成分含量最高。UV-Vis FP,HPLC FP结合主成分分析和聚类分析能快速鉴别不同采收期灯台叶并对其进行品质评价。灯台叶最佳采收期为10月至次年2月,即傣历中的冷季。UV-Vis and HPLC fingerprint of different harvest time of the leaves of Alstonia scholaris（L.）R.Br.were establish the for identification and quality evaluation to promote the development of Dai Medicine modernization.The optimal extraction condition was used to obtain UV-vis data of different harvest time which were deducted background and eight spot smooth,were collected to make the principal component analysis in SIMCA-P＋11.5,identifying the samples quickly with the first three principal component three-dimensional diagram.The HPLC fingerprint were obtained with Agilent ZORBAX Eclipse XDB C18（250×4.6mm,5μm）chromatographic column with the mobile phase of acetonitrile（B）-water（contain 0.1% formic acid）（A）for gradient elution（0~5min,5% B;5~35min,5% B→26% B;35~40min,26% B→56% B）.The wavelength was set at 287 nm and the column temperature was maintained at 30 ℃.The flow rate was 1.0mL·min^（-1） and the injection volume was 7μL.The HPLC fingerprint of different harvest time of the leaves of Alstonia scholaris（L.）R.Br.was analysised by cluster analysis to quality evaluation.Research findings showing：（1）The UV-Vis spectrogram of different harvest time of the leaves of Alstonia scholaris（L.）R.Br.were divided into three parts according to the absorption peak position and amplitude of variation.The first was 235 to 400nm,the second was 400 to 500nm,and the third was 500 to 800nm.In the first part,absorption peak were focused on 270,287 and 325nm,which can reflect the fingerprint character for the high absorbance and amplitude of variation.Absorption peak were distributed in 410 and 464nm in the second part,absorbance and amplitude of variation were lower than the first part.There was a bigger absorption peak at 665 nm in the third part,but the absorbance had no difference.The UV-Vis data of different harvest time were gathered to make the principal component analysis,the result was that the samples of same month were concentrated distribution,but different month samples

关 键 词：紫外-可见光谱 HPLC指纹图谱 傣药 灯台叶 采收期 鉴别 质量评价 

分 类 号：O657.3[理学—分析化学]