陈明伟 韦锦梅 甄丹丹 丘琴 卢显兴 黄秋艳 甄汉深

摘要：【目的】以廣西产罗汉松[Podocarpus macrophyllus （Thunb.） D. Don]的叶为研究对象，建立高效液相色谱（HPLC）指纹图谱，为罗汉松叶的质量评价提供参考依据。【方法】利用HPLC对广西产罗汉松叶进行指纹图谱研究，建立共有模式，并进行方法学考察，使用相似度评价系统对10批不同产地罗汉松叶的共有峰进行确认，对其相似度进行评价;使用SPSS 21.0，以共有峰原始峰面积数据进行聚类分析。【结果】方法学考察结果中，精密度试验主要共有峰的相对保留时间和相对峰面积的相对标准偏差（RSD）均小于3%;稳定性试验相对峰面积的RSD均小于3%，表明样品提取液在24 h内较稳定;重复性试验所有色谱峰的相对保留时间的RSD均小于2%;建立罗汉松叶的指纹图谱，10批不同产地罗汉松叶初步建立其共有模式，标定了22个共有峰，样品指纹图谱与对照指纹图谱相似度都在0.900以上，相似性良好。在10批罗汉松叶药材的指纹图谱中，共有峰的相对保留时间无明显差异。10批罗汉松叶样品可分为3类：S4为第1类;S3、S2和S1为第2类;S5、S6、S7、S8、S9和S10为第3类。【结论】建立的罗汉松叶指纹图谱特征性强，准确可靠，可为罗汉松叶的质量评价提供参考依据。

关键词： 罗汉松叶;质量分析;指纹图谱;高效液相色谱（HPLC）

中图分类号： S791.46;R284.1                          文献标志码： A 文章编号：2095-1191（2021）08-2078-09

Fingerprint of Podocarpus macrophyllus（Thunb.） D. Don

leaves from Guangxi

CHEN Ming-wei， WEI Jin-mei， ZHEN Dan-dan， QIU Qin*， LU Xian-xing，

HUANG Qiu-yan， ZHEN Han-shen

（Guangxi University of Chinese Medicine， Nanning  530200， China）

Abstract：【Objective】The high efficiency liquid chromatography（HPLC） fingerprint was adopted to analyze leaves of Podocarpus macrophyllus（Thunb.） D. Don. from Guangxi，so as to provide a reference for the quality evaluation of leaves of P. macrophyllus. 【Method】HPLC was used to study the fingerprint of P. macrophyllus leaves and the common fingerprint mode was established，the method validation was carried. The similarity evaluation system for chromatographic fingerprint of TCM（2012.1 edition） was used to calculate the similar degree of common peaks of 10 batches of P. macrophyllus leaves from different places of origins. The SPSS 21.0 statistical software was used to cluster the original peak area data of the common peaks. 【Result】In precision tests，relative standard deviation（RSD） of relative retention time and relative peak area of the major common peaks were less than 3%. RSD of relative peak area in stability tests are all less than 3%，the results showed that the sample extract was stable within 24 h. RSD of the relative retention time of all chromatographic peaks in the reproducibility tests were less than 2%. The fingerprints of the leaves of P. macrophyllus were established. The common fingerprint pattern of 10 batches of P. macrophyllus leaves were initially established，22 common peaks were confirmed. The similarity between the fingerprint of the sample and the control was more than 0.900，the similarity was good. There was no great difference in the relative retention time of the common peaks among 10 batches of P.macrophyllus leaves. The 10 batches of P. macrophyllus leaves could be divided into 3 categories. S4 into the first category，S3，S2 and S1 were grouped into the second category，S5，S6，S7，S8，S9 and S10 were grouped into the third category.【Conclusion】The fingerprint established is characterized，accuracy and reliability，which can provide basis for the quality evaluation of the P. macrophyllus leaves.

Key words： Podocarpus macrophyllus（Thunb.） D. Don leaves; quality analysis; fingerprint; high efficiency liquid chromatography（HPLC）

Foundation item： Guangxi First-class Discipline Construction Project（2018XK052）; Zhuang and Yao Pharmaceutical Collaborative Innovation Center Project（Guijiaokeyan〔2013〕20）; Guangxi Zhuang and Yao Medicine Key Laboratory Project（Guikejizi〔2014〕32）; Guangxi Key Discipline Zhuang Pharmacy Project（Guijiaokeyan〔2013〕16）

0 引言

【研究意义】罗汉松[Podocarpus macrophyllus（Thunb.） D. Don]又名羅汉柏、罗汉杉、金钱松、长青罗汉杉、土杉、仙柏等，为罗汉松科罗汉松属植物，分布在热带、亚热带和南温带地区，在广西地区常见。罗汉松既可作观赏植物，又可作药用植物，其全株均可药用（吴鲁东等，2012;张东峰，2020），其中穗花杉双黄酮为罗汉松的主要活性成分（黄振青等，2014;杨雨等，2014）。……