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[利用超高效液相色谱法和液相色谱-质谱/质谱联用技术分析丹参根和云南鼠尾草根中的亲脂性成分]

[Analysis of lipophilic components of Salvia miltiorrhiza roots and S. yunnanensis roots by UPLC and LC-MS/MS].

作者信息

Zhang Li-Guo, Hu Tian-Tian, Zhang Fang-Fang, Luan Shao-Rong, Li Wei, Deng Hai-Xing, Lan Zhu-Hui, Luo Xiao-Fang, Wu Zhong-Xiang, Leslaw Mleczko

机构信息

School of Chemistry & Molecular Engineering,East China University of Science and Technology Shanghai 200237,China.

Sichuan Dian Hong Pharmaceutical Development Co.,Ltd. Chengdu 610000,China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2019 Mar;44(6):1208-1215. doi: 10.19540/j.cnki.cjcmm.20181226.020.

Abstract

Fingerprints of lipophilic components in the roots of Salvia miltiorrhiza and S.yunnanensis were analyzed by UPLC-DADand UPLC coupled with mass spectroscopy to evaluate the differences and similarities of the lipophilic components in the two kinds of herbs.The UPLC analysis of 18 batches of S.miltiorrhiza and 16 batches of S.yunnanensis was performed on a 25℃Thermo Accucore C_(18)column(2.1 mm×100 mm,2.6μm)by Shimadzu LC-20AD;mobile phase was 0.026%phosphoric acid(A)-acetonitrile(B)with gradient elution;flow rate was 0.4 m L·min~(-1);detection wavelength was set at 270 nm;injection volume was 2μL.The molecular structures of the lipophilic components were analyzed on a 25℃Thermo Accucore C_(18)column(2.1 mm×100 mm,2.6μm)by Thermo U3000 UPLC Q Exactive Orbitrap LC-MS/MS with a mobile phaseconsisting of 0.1%formic acid water(A)and 0.1%formic acidacetonitrile(B).The mass spectrometry was acquired in positive modes using ESI.There are 10 common peaks in the lipophilic components of S.miltiorrhiza.The similarity between the 16 batches of S.miltiorrhiza and their own reference spectra was greater than 0.942,and the average similarity was 0.973.There are 12 common peaks in the lipophilic components of S.yunnanensis.The similarity between the 18 batches of S.yunnanensis and their own reference spectra was greater than 0.937,and the average similarity was 0.976.The similarity between the reference chromatograms of S.miltiorrhiza and S.yunnanensis was only 0.900.There are three lipophilic components in S.yunnanensis,which are not found in S.miltiorrhiza,and one of which isα-lapachone.There is a lipophilic component in S.miltiorrhiza not found in S.yunnanensis,which may be miltirone.The two herbs contain 8 common lipophilic components including dihydrotanshinoneⅠ,cryptotanshinone,tanshinoneⅠ,tanshinoneⅡ_A,nortanshinone in which the content of tanshinoneⅡ_A,dihydrotanshinoneⅠand cryptotanshinone of S.yunnanensisis significantly lower than that of S.miltiorrhiza(P<0.01),and the contents of tanshinoneⅠand nortanshinone are significantly lower than that of S.miltiorrhiza too(P<0.05).There are significant differences in the types and contents of lipophilic components between the roots of S.miltiorrhiza and S.yunnanensis,and the similarity between the fingerprints of interspecies is much lower than that between the same species.Therefore,the roots of S.miltiorrhiza and S.yunnanensis are two kinds of herbs which are quite different in chemical compounds and compositions.

摘要

采用超高效液相色谱-二极管阵列检测法(UPLC-DAD)和超高效液相色谱-质谱联用技术,对丹参和云南鼠尾草根部的亲脂性成分指纹图谱进行分析,以评价这两种药材中亲脂性成分的异同。采用岛津LC-20AD,在25℃的Thermo Accucore C₁₈柱(2.1 mm×100 mm,2.6μm)上对18批丹参和16批云南鼠尾草进行UPLC分析;流动相为0.026%磷酸(A)-乙腈(B),梯度洗脱;流速为0.4 mL·min⁻¹;检测波长设定为270 nm;进样量为2μL。采用Thermo U3000 UPLC Q Exactive Orbitrap LC-MS/MS,在25℃的Thermo Accucore C₁₈柱(2.1 mm×100 mm,2.6μm)上分析亲脂性成分的分子结构,流动相由0.1%甲酸水(A)和0.1%甲酸乙腈(B)组成。质谱采用电喷雾电离正离子模式采集。丹参亲脂性成分中有10个共有峰。16批丹参与其自身对照图谱的相似度大于0.942,平均相似度为0.973。云南鼠尾草亲脂性成分中有12个共有峰。18批云南鼠尾草与其自身对照图谱的相似度大于0.937,平均相似度为0.976。丹参和云南鼠尾草对照色谱图之间的相似度仅为0.900。云南鼠尾草中有3种亲脂性成分在丹参中未发现,其中一种为α-拉帕醌。丹参中有1种亲脂性成分在云南鼠尾草中未发现,可能是丹参酮。两种药材含有8种共同的亲脂性成分,包括二氢丹参酮Ⅰ、隐丹参酮、丹参酮Ⅰ、丹参酮ⅡA、去甲丹参酮,其中云南鼠尾草中丹参酮ⅡA、二氢丹参酮Ⅰ和隐丹参酮的含量显著低于丹参(P<0.01),丹参酮Ⅰ和去甲丹参酮的含量也显著低于丹参(P<0.05)。丹参和云南鼠尾草根部亲脂性成分的种类和含量存在显著差异,种间指纹图谱的相似度远低于种内相似度。因此,丹参和云南鼠尾草是两种化学成分和组成差异较大的药材。

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