采用HPLC法建立了花椒的高效液相指纹图谱。色谱条件为色谱柱:迪马铂金C18(250 mm×4.6 mm,5μm),柱温:30℃,流动相:V(乙腈)∶V(水)=50∶50,流速:0.8 mL/min,检测波长:254 nm,进样量2μL,检测时间:40 min。采用《中药色谱指纹图谱相似度评价系统A版》(研究版)提取HPLC图谱共有模式,计算各批次样品的相似度,结果显示,红花椒特征指纹图谱中有11个共有峰,15批红花椒样品中,除一份样本外,其他14批样品相似度均大于0.97,表明不同产地红花椒内在品质相似;青花椒特征指纹图谱中有10个共有峰,18批青花椒样品相似度都很低,相似度为0.52～0.67,表明不同产地青花椒内在品质差异性很大。红花椒相比青花椒在保留时间约30 min处存在一个比较明显的色谱峰,并采用飞行时间质谱对该特征差异色谱峰进行鉴定,推断该物质为不饱和五烯酰胺,即羟基-γ-山椒素或2'-羟基-N-异丁基-2,4,8,10,12-十四烷五烯酰胺及其同分异构体。

HPLC fingerprint of Pericarpium Zanthoxyli were established by fingerprinting technology.Platisil ODS C18 Column(250 mm×4.6 mm,5 μm)was used with acetonitrile-water(50∶50) as mobile phase at a flow rate of 0.8 mL/min.The detection wavelength was at 254 nm.The column temperature was at 22 ℃.The characteristic of Pericarpium Zanthoxyli fingerprint was established by《Evaluation Systems of similarity of the chromatographic fingerprint of traditional Chinese medicine,version A》(researchful version).Fingerprint of Zanthoxylum Bungeanum Maxim has eleven common peaks,fingerprint of Zanthoxylum Schinifolium Sieb.et Zucc has ten common peaks.Referring to common peaks,similarity value of Zanthoxylum Bungeanum Maxim and Zanthoxylum Schinifolium Sieb.et Zucc was calculated.The results showed fingerprint similarity of Zanthoxylum Bungeanum Maxim was high(amount similarity>0.97),but fingerprint of Zanthoxylum Schinifolium Sieb.et Zucc was low similarity(0.52~0.67).Quality of Zanthoxylum Bungeanum Maxim from different origins was more consistent,while Zanthoxylum Schinifolium Sieb.et Zucc has big difference.Characteristic difference between peak of HPLC fingerprint of Zanthoxylum Bungeanum Maxim and Zanthoxylum.Schinifolium Sieb.et Zucc was identified by TOF/MS,and was inferred to hydroxyl-γ-sanshool or 2'-hydroxyl-N-isobutyl-2,4,8,10,12-tetradepentaenoateamide or their structural isomer.