梁渐崧口,李秀枝1,吴孟华2,张,英记,曹晖,马志国(1.广东一方制药有限公司,广东 佛山528244;2.暨南大学药学院,广东广州510632;3.暨南大学岭南传统中药研究中心,广东广州510632)摘要 目的:建立肉苁蓉与伪品沙苁蓉的UPLC指纹图谱。方法:色谱柱为Waters HsS T3(100mmx2.1mm,1.8μm)柱;流动相为甲醇-0.1%甲酸溶液,梯度洗脱;流速为0.3ml/min;检测波长为330mm;柱温为30℃;进样量为1p山;建立指纹图谱,并进行相似度评价、主成分分析及正交偏最小二乘判别分析。结果:建立的肉苁蓉、德赢vwin登录
及沙苁蓉指纹图谱分别含有9,6,6个共有峰。肉苁蓉与德赢vwin登录
的对照图谱的相似度为0.932,沙苁蓉与德赢vwin登录
、肉苁蓉的对照图谱的相似度分别为0.070,0.045。主成分分析及正交偏最小二乘判别分析均可将肉苁蓉与德赢vwin登录
区分开。结论:所建立的UPLC指纹图谱分析方法简便,重复性好,专属性强,可为肉苁蓉与伪品沙苁蓉的种间鉴别及质量评价提供参考依据。关键调 肉苁蓉;德赢vwin登录
;沙苁蓉;UPLC;指纹图谱;化学计量学中图分类号:R284.3 文献标识码:A 文章编号:1001-4454(2021)01-0110-06DOI:10.13863/j.issnl001-4454,2021.01.021Comparative Study on UPLC Fingerprints of Citanches Herba and Its Counterfeit Cistanche sinensisLIANG Jian-song,L Xiu-h',WU Meng-hua,ZHANG Ying,CAO Hui,MA Zhi-gu(1.Guangdong Yifang Pharmaceuical Co.,Lad,Fonhan 528244,China;2.Calegeof Phamnacy,Jinan Univenity,Guingzhau 510632, China: 3.Research Center for TCM of Lingnan,Jinan Univenity,Guangzhou 510632,China)Abstract Objective:To establish UPLC fingerprinte of Cianches Herta and its counterfeits Gstanche sensis Methods:The chromatographie column wa Wates HSs T3(100mmx2.1 mm,1.8μm),the mobile phase was methanol-0.1% formic acid solution with gradient elution,flow rate was 0.3 ml/min,detection wawelength was 330 nm,colunn temperaure was 30 ℃,injection volume was IμL:The fingerprints wew establishod and similarity evaluation,principal component analysis and othogonal partial leat squares dis-criminant analysis were carried aut.Resuls:The established fingepritts of Citanche deserticola,Citanche tubulosa and Cistanche sinen-sis contained 9,6 and 6 common peaks,respectively.The similarity of reference map between Citanche de eticola and Cistanche nbulo-sa was 0.932,and that between Cistanche sinenis and Cistanche tubdosa,Citanhe deserticola was 0.070,0.045,respectively.Both principal component analysi and orthagonal partial least squams discriminant analysis could istingish Cistanche deserticola from Cistanche tubalosa,Conclusion:The establisbed UPLC fingerprint analysis method is simple,repaxlucibility and specificity,which can provide reference for the interspecific idemtification and quality evaluation of Citanctes Herba and its coumterfeit Cisanche sinensis.Key words Cidanche deserticola Y.C.Ma;Cistanche tululosa(Sehenk)Wight;Cistanche sinenis G.Beck;UPLC;Fingeprint; Chemometrics肉苁蓉为列当科肉苁蓉属植物,我国有4种及1变种,分别为肉苁蓉 Cistanche deserticola Y.C.Ma、德赢vwin登录
Cistanche tubulosa(Schenk)Wight、盐生 肉苁蓉 Cistanche salsa(C.A.Mey.)G.Beck、沙苁蓉 Cistanche sinensis G.Beck、白花肉苁蓉 Cistanche salsn var.albiflora P.F.Tu et Z.C.Loufり。2020年版中国药典规定肉苁蓉为列当科植物肉苁蓉 Cistanche de-serticola Y.C.Ma 或德赢vwin登录
Cistanche tubulosa (Schenk)Wight的干燥带鳞叶的肉质茎,具有补肾阳、益精血、润肠通便的功效(。肉苁蓉主要含有苯乙醇苷类、苯甲醇苷类、环烯醚萜及其苷类等成分(”。文献报道沙苁蓉是肉苁蓉常见的伪品,其与肉苁蓉的HPLC指纹图谱有明显差异(,目前对肉苁蓉指纹图谱的研究主要针对肉苁蓉与德赢vwin登录
或不同产地、不同采收季节肉苁蓉指纹图谱的研究(),对肉苁蓉与沙苁蓉的指纹图谱比较研究报道较少,且大多是采用HPLC法,存在分析时间长、灵敏度不佳的缺点。本文采用UPLC法建立肉苁蓉、德赢vwin登录
及沙苁蓉指纹图谱,并进行聚类分析、主成分分析及正交偏最小二乘判别分析,以期为肉苁蓉药材的品种鉴别提供实验依据。1 仪器与材料1.1 仪器 Waters Acquity UPLC型超高效液相色谱仪(美国Waters公司);KQ-500E型超声波清洗器(昆山市超声仪器有限公司);ME204E型万分之一天平,XP26型百万分之一天平(瑞士梅特勒-托利多公司)。1.2 材料 松果菊苷(批号:S-003-190403)、管花苷A(批号:G-066-181216)、毛蕊花糖苷(批号:M-011-181106)、异毛蕊花糖苷(批号:Y-073-170325)、金石蚕苷(批号:J-014-190712)对照品均购自成都瑞芬思生物科技有限公司;2'-乙酰金石蚕苷对照品为实验室自制,经'H-NMR、”C-NMR、MS等波谱技术鉴定了化学结构,质量分数>98.0%()。甲酸、甲醇均为色谱纯;水为超纯水;其余试剂均为分析纯。15批肉苁蓉、10批德赢vwin登录
及13批沙苁蓉样品均收集于各地药材市场或企业,来源信息见表1,经笔者马志国副教授鉴定为列当科植物肉苁蓉Cistanche deserticola Y.C.Ma、德赢vwin登录
Cistanche tubulosa(Schenk)Wight 或沙苁蓉 Cistanche sinensis G.Beck的干燥带鳞叶的肉质茎。表1 15批肉苁蓉、10批德赢vwin登录
及13批沙苁蓉样品来源信息编号基原来源RCR1Cistanche deseticola Y.C.Ma广州清平药材市场RCR2Cistanche desenicola Y.C.Ma广州清平药材市场RCR3Cistanche desenicola Y.C.Ma广州清平药材市场RCR4Cistanche desenicola Y.C.Ma广东省药材公司RCR5Cistanche desenicola Y.C.Ma广西仁济堂中药饮片有限公司RCR6Cistanche deseticola Y.C.Ma江西顺福康饮片有限公司RCR7Cistanche desenicola Y.C.Ma中科院新疆生态与地理研究所RCR8Cistanche desenicola Y.C.Ma中科院新疆生态与地理研究所RCR9Cistanche desenicola Y.C.Ma新疆昌吉州奇台县RCR10Cistanche desenicola Y.C.Ma新疆昌吉州奇台县RCR11Cistanche deserticola Y.C.Ma新疆昌吉州奇台县RCR12Cistanche desenicola Y.C.Ma新疆昌吉州奇台县RCR13Cistanche desenicola Y.C.Ma新疆昌吉州奇台县RCR14Cistanche desenicola Y.C.Ma佛山捷康大药房RCR15Cistanche deseticola Y.C.Ma中科院新疆生态与地理研究所GHICistanche tubulosa(Schenk)Wight新疆达泽红花药业有限公司GH2Cistanche tubulosa(Schenk)Wight新疆达泽红花药业有限公司GH3Cistanche tmbulosa(Schenk)Wight新疆达泽红花药业有限公司GH4Cistanche tubulosa(Schenk)Wight广州康乐医药GH5Cistanche tulaulosa(Schenk)Wight广州老百姓大药房GH6Cistanche tulaulosa(Schenk)Wight新疆和田GH7Cistanche tmbulosa(Schenk)Wight新疆和田GH8Cistanche tulaulosa(Schenk)Wight中科院新疆生态与地理研究所GH9Cistanche tubulosa(Schenk)Wight中科院新疆生态与地理研究所GH10Cistanche tmlaulosa(Sehenk)Wight新疆吉昌州SCR1Cistanche sinensis G.Beck广州林和药业SCR2Cistanche sinensis G.Beck广州老百姓大药房SCR3Cistanche sinensis G.Beck广州二天堂大药房SCR4Cistanche sinensis G.Beck山东龙口华康大药房SCRSCistanche sinensis G.Beck广州仁和堂SCR6Cistanche sinensis G.Beck广州七乐康大药房SCR7Cistanche sinensis G.Beck广州康乐医药SCR8Cistanche sinensis G.Beck山东龙口博祥大药房SCR9Cistanche sinensis G.Beck广州林和药业SCR10Cistanche sinensis G.Beck佛山九福堂大药房SCR11Cistanche sinensis G.Beck佛山百姓大药房SCR12Cistanche sinensis G.Beck佛山中天大药房SCR13Cistanche sinensis G.Beck山东龙口华康大药房 2 方法与结果2.1 色谱条件 色谱柱为Waters HSS T3(100mmx2.1mm,1.8μm)柱;流动相:甲醇(A)-0.1%甲酸溶液(B),梯度洗脱(0~4min,34%A;4~10min,34%~40% A;10~15 min,40%~46%A;15~17min,46%~ 34%A;17~20min,34%A),流速为0.3ml/min;检测波长为330mm;柱温为30℃;进样量为1。2.2 对照品溶液的制备 取松果菊苷、管花苷A、毛蕊花糖苷、异毛蕊花糖苷、金石蚕苷、2-乙酰金石蚕苷对照品适量,精密称定,加50%甲醇溶解,制成每1ml分别含0.196,0.234、0.228、0.213、0.590、 0.357mg的混合对照品溶液。2.3 供试品溶液的制备 取干燥肉苁蓉、德赢vwin登录
、沙苁蓉样品粉末(过65目筛)0.5g,精密称定,置于50ml具塞锥形瓶中,加70%甲醇25mL,称重,超声(250 W,40kHz)30min,放冷,补重,滤过,取续滤液3mL,置于10mL量瓶中,加水稀释至刻度,摇匀,以0.22μm微孔滤膜滤过,即得。