不同产地、不同部位荒漠肉苁蓉的指纹图谱特征


摘 要

中药材在我国具有悠久的用药历史，是传统医学的物质基础，为治疗疾病和维持身体健康发挥了不可替代的重要作用。我国道地药材资源丰富且疗效显著。中药指纹图谱是评价中药材药材道地性的有效方法之一，常用于中药的质量评价、质量控制和新药研究等，为推进我国中医药行业的长足发展提供了有力支撑。

本文以荒漠肉苁蓉为研究对象，通过液相色谱技术与傅里叶中红外光谱技术进行检测，分别建立了不同产地和不同部位的液相色谱指纹图谱与中红外光谱指纹图谱；结合化学计量学方法对荒漠肉苁蓉的苯乙醇苷成分与中红外光谱信息进行了分析；对不同产地荒漠肉苁蓉醇提物与粗多糖的抗氧化活性进行评价，旨在为荒漠肉苁蓉的药材鉴别与品质评价提供基础数据。通过研究，得出以下主要结果：

（1）建立了新疆、内蒙古和哈萨克斯坦产地荒漠肉苁蓉高效液相色谱指纹图谱，其中内蒙古荒漠肉苁蓉苯乙醇苷成分以松果菊苷和毛蕊花糖苷为主，且含量高于新疆与哈萨克斯坦产地样品。新疆与哈萨克斯坦产地荒漠肉苁蓉苯乙醇苷成分以松果菊苷和2′-乙酰毛蕊花糖苷为主。主成分分析结果表明，其可用于不同产地荒漠肉苁蓉的产地鉴别。

（2）建立了荒漠肉苁蓉花序部位与茎部位的高效液相色谱指纹图谱，荒漠肉苁蓉花序部位中苯乙醇苷成分以2'-乙酰毛蕊花糖苷为主，未检测出肉苁蓉苷A和异毛蕊花糖苷；荒漠肉苁蓉茎部位中苯乙醇苷成分以松果菊苷、毛蕊花糖苷和2'-乙酰毛蕊花糖苷为主，7种苯乙醇苷成分均有检出。综合评价结果显示，茎部位的综合品质高于花序部位。

（3）建立了新疆、内蒙古、青海和哈萨克斯坦产地荒漠肉苁蓉中红外光谱指纹图谱。不同产地中红外光谱的差异性分析表明内蒙古、新疆、青海及哈萨克斯坦样品分别具有14、14、17和16个特征吸收峰，共确定了9个共有特征吸收峰。通过光谱谱峰归属分析，判断出荒漠肉苁蓉中含有苯乙醇苷类、多糖类等成分。不同产地中红外光谱分析中发现，1 750~700 cm-1为一维中红外光谱指纹谱区，1 100~850 cm-1为二阶导数谱图指纹谱区，不同产地荒漠肉苁蓉具有区别于其他产地的吸收特征峰。

（4）建立了基于SIMCA-P的PCA（主成分分析）产地鉴别模型，其中R2X为0.777，Q2为0.723，所建模型的拟合度较好，预测能力较强，对于不同产地荒漠肉苁蓉样品区分效果良好。对不同产地荒漠肉苁蓉的共有峰率和变异峰率双指标序列分析表明，地理位置接近的样品之间共有峰率较高，变异峰率较低；反之，则共有峰率较低，变异峰率较高。

（5）建立了荒漠肉苁蓉花序部位与茎部位的中红外光谱指纹图谱，共确定12个吸收峰作为不同部位的共有特征吸收峰，建立荒漠肉苁蓉不同部位的OPLS-DA鉴别模型，其中R2Y为0.749，Q2Y为0.616，所建模型拟合度和预测结果较好。综合评价分析结果表明茎部位的综合品质高于花序部位。

（6）通过体外抗氧化活性测定分析发现，不同产地荒漠肉苁蓉醇提物和粗多糖对DPPH和ABTS自由基均具有良好的清除效果。经TOPSIS综合分析发现，醇提物的综合抗氧化能力从大到小依次为：青海样品＞内蒙古样品＞新疆样品＞哈萨克斯坦样品。粗多糖的综合总抗氧化能力从大到小依次为：哈萨克斯坦样品＞内蒙古样品＞新疆样品＞青海样品。

关键词：荒漠肉苁蓉，指纹图谱，高效液相色谱，中红外光谱，抗氧化活性

Abstract

Chinese herbal medicine has a long history of medication in China.It is the material basis of traditional medicine and plays anirreplaceable role in treating diseases and maintaining health. There are many authentic medicinal materials with significant curative effect and rich resources in China. Fingerprint of traditional Chinese medicine is one of the methods to evaluate the authenticity of authentic medicinal materials. It can be used for quality evaluation,quality control and new drug researchof traditional Chinese medicine, which can provide a strong development basis for promoting the basic research of traditional Chinese medicine industry in China.

In this paper, the Cistanche deserticola were selected as the research object, the HPLC and Fourier transform infrared spectroscopy (FTIR) were usedto establish the fingerprints of different areas and different parts of C. deserticola.The phenylethanoid glycosides and mid infrared spectrum information of C.deserticola were analyzed by chemometrics method.At the same time, the antioxidant activities of ethanol extract and crude polysaccharide of C.deserticola from different areas were evaluated.The purpose was to provide a theoretical basis for the identification and quality evaluation of C.deserticola industry.Through the study, the main results were obtained as follows :

(1) HPLC fingerprints of C.deserticola from Xinjiang, Inner Mongolia and Kazakhstan were established.The main components of phenylethanolic glycosides in C.deserticola from Inner Mongolia were echinacoside and verbascoside,and the contents were higher than those from Xinjiang and Kazakhstan. Echinacoside and 2'-Acetylacteoside were the main phenylethanoid glycosides in C.deserticola from Xinjiang and Kazakhstan.The results of principal component analysis showed that it could be used to identify the producing areas of C. deserticola from different producing areas.

(2) HPLC fingerprints of inflorescence and stem of C.deserticola were established.2'-acetylverbascoside was the main phenylethanoid glycoside in inflorescence of C.deserticola, but the Cistanoside A and Isoacteoside were not detected.Echinacoside, Acteoside and 2'-Acetylacteoside werethe main phenylethanoid glycosides in the stem of C. deserticola, and seven phenylethanoid glycosides were detected in the stem parts.Comprehensive evaluation results showed that the comprehensive quality of stem was higher than that of inflorescence.

(3) The mid infrared spectrum fingerprints of C.deserticola from different producing areas was established.The samples from Xinjiang, Inner Mongolia, Qinghai and Kazakhstan had 14, 14,17 and 16 characteristic absorption peaks respectively, and 9 common characteristic absorption peaks were identified. According to the spectral peak assignment analysis,it was found that there were phenylethanoid glycosides and polysaccharides in C. deserticola.The results showed that 1 750-700 cm-1 was one-dimensional mid infrared spectrum fingerprint area, and 1 100-850 cm-1 was second derivative spectrum fingerprint area. C.deserticola from different habitats had absorptioncharacteristic peaks different from other habitats.

(4) The PCA (principal component analysis) origin identification model based on SIMCA-P was established, the r2x was 0.777 and the Q2 was 0.723.The model had good fitting degree and strong prediction ability, and could distinguish C.deserticola samples from different origins.Sequence analysis of common peak rate and variation peak rate of C.deserticola from different habitats showed that the common peak rate was higher and the variation peak rate was lower among samples in close geographical position;on the contrary, the common peak rate was lower and the variation peak rate was higher

(5) The mid infrared spectrum fingerprints of inflorescence and stem of C.deserticola were established.A total of 12 absorption peaks were identified asthe common characteristic peaks of different parts. The OPLS-DA identification model of different parts of C.deserticola was established, the r2y was 0.749 and q2y was 0.616.The prediction results of the model were good,which could effectively distinguish different parts of C. deserticola.The results of comprehensive evaluation and analysis showed that the comprehensive quality of stem was higher than the inflorescence.

(6) Through in vitro antioxidant activity determination and analysis,it is found that the ethanol extract and crude polysaccharide of Cistanche deserticola from different areas had good scavenging effects on DPPH and ABTS free radicals. TOPSIS analysis showed that the order of antioxidant capacity of ethanol extractwas Qinghai sample > Inner Mongolia sample > Xinjiang sample > Kazakhstan sample. The order of total antioxidant capacity of crude polysaccharides from large to small was Kazakhstan > Inner Mongolia > Xinjiang > Qinghai.

Key Words: Cistanche deserticola, Fingerprint, HPLC,Mid infrared spectrum, Antioxidant activity