维吾尔药材菠菜子的DNA条形码鉴定及质量控制研究

目的:基于DNA条形码分子鉴定法建立维吾尔药材菠菜子的鉴定方法,并构建菠菜子HPLC指纹图谱和其指标性成分β-蜕皮甾酮的HPLC含量测定方法,为菠菜子的质量控制提供参考。方法:以核糖体DNA第二内部转录间隔区(ITS2)为主体条形码序列对菠菜子进行DNA条形码鉴定;采用Agilent ZORBAX SB-Aq C_(18)色谱柱(4.6 mm×250 mm,5μm),以甲醇(A)-0.2%磷酸溶液(B)为流动相梯度洗脱(0~10 min,10%A;10~20 min,10%→30%A;20~40 min,30%→40%A;40~65 min,40%→80%A),检测波长240 nm,建立菠菜子的HPLC指纹图谱方法;采用Agilent ZORBAX SB-Aq C_(18)色谱柱(4.6 mm×250 mm,5μm),以甲醇(A)-0.2%磷酸溶液(B)为流动相梯度洗脱(0~15 min,45%→40%A;15~25 min,40%→80%A;25~30 min,80%→45%A;30~45 min,45%A),检测波长248 nm,建立指标性成分β-蜕皮甾酮的HPLC含量测定方法。结果:DNA条形码鉴定系统进化树显示,ITS2序列可将菠菜子与其近缘物种很好区分,且分支支持率达100%;HPLC指纹图谱标定7个共有峰,10批菠菜子药材与对照图谱的相似度均大于0.95;指标性成分β-蜕皮甾酮在6.115~122.304μg/mL范围内线性关系良好(r=0.9997),重复性试验RSD<2.0%,加样回收率为97.43%~103.59%,10批样品中β-蜕皮甾酮的含量为0.099%~0.241%。结论:DNA条形码可准确鉴定菠菜子的基原,HPLC指纹图谱专属性强,结合指标性成分β-蜕皮甾酮的含量测定可以控制菠菜子药材的质量。

Inorganic Elemental Determinations of Marine Traditional Chinese Medicine Meretricis concha from Jiaozhou Bay:The construction of Inorganic Elemental Fingerprint Based on Chemometric Analysis

The goal of this paper is to explore the relationship between the inorganic elemental fingerprint and the geographical origin identification ofMeretricis concha, which is a commonly used marine traditional Chinese medicine （TCM） for the treatment of asthma and scald bums. For that, the inorganic elemental contents ofMeretricis concha from five sampling points in Jiaozhou Bay have been determined by means of inductively coupled plasma optical emission spectrometry, and the comparative investigations based on the contents of 14 inorganic elements （A1, As, Cd, Co, Cr, Cu, Fe, Hg, Mn, Mo, Ni, Pb, Se and Zn） of the samples from Jiaozhou Bay and the previous reported Rushan Bay were performed. It has been found that the samples from the two bays are ap- proximately classified into two kinds using hierarchical cluster analysis, and a four-factor model based on principle component analysis could explain approximately 75% of the detection data, also linear discriminant analysis can be used to develop a prediction model to distinguish the samples from Jiaozhou Bay and Rushan Bay with accuracy of about 93%. The results of the present investi- gation suggested that the inorganic elemental fingerprint based on the combination of the measured elemental content and chemom- etric analysis is a promising approach for verifying the geographical origin ofMeretricis concha, and this strategy should be valuable for the authenticity discrimination of some marine TCM.