目的: 建立能有效区分硬尖神香草Hyssopus cuspidatus Boriss.及其混淆品大苞荆芥Nepeta bracteata Benth.和欧神香草Hyssopus officinalis L.的指纹图谱和一测多评（QAMS）法,为硬尖神香草的真伪鉴别和质量评价提供依据。方法: 采用Agilent XDB C18（250 mm×4.6 mm,5 μm）色谱柱,以乙腈（A）-0.1%甲酸溶液为流动相,梯度洗脱,流速1.0 mL · min-1,柱温30 ℃,检测波长280 nm,进样量20 μL。对硬尖神香草中化学成分进行分析,建立指纹图谱方法,并结合相似度评价、聚类分析（CA）、主成分分析（PCA）、正交偏最小二乘法-判别分析（OPLS-DA）法,筛选确认差异性质量标志物;采用熵权优劣解距离（TOPSIS）评价不同批次及产地的硬尖神香草及其混淆品,筛选最佳产地;以迷迭香酸为内参物,建立了9个差异性质量标志物的QAMS。结果: 建立了22批药材的指纹图谱,共标识51个共有峰,通过与对照品比对指认出15个色谱峰（新绿原酸、绿原酸、咖啡酸、阿魏酸、槲皮素-3-O-β-D吡喃葡萄糖苷酸、迷迭香酸、丹酚酸B、木犀草素、迷迭香酸甲酯、迷迭香酸乙酯、β-谷甾醇、芫花素、山柰素、三裂鼠尾草素、山楂酸）,样品与对照图谱相似度在0.509~0.996;CA将22批神香草样品分为3大类;PCA不同硬尖神香草及其混淆品化学成分存在一定的差异,并提取区别硬尖神香草及混淆品质量的主成分5个;OPLS-DA筛选出12个可能的差异性质量标志物;同时建立9个差异性质量标志物的QAMS。9个差异性质量标志物的线性关系均良好（r≥0.999 0）,精密度、稳定性、重复性和耐用性试验测定结果的RSD均<3.0%,平均加样回收率97.5%~103.1%,RSD均<2.0%。以迷迭香酸为内参物建立的QAMS与外标法（ESM）的含量测定结果比较,P分别为0.959、0.964、0.971、0.988、0.999、0.889、0.962、0.972、0.997,均无显著性差异（P>0.05）。结论: 建立的指纹图谱和一测多评方法简便可行,专属性强,建立的质量综合评价模式全面客观,可用于硬尖神香草及其混淆品药材的真伪鉴别和质量评价。
Objective: To establish the HPLC fingerprint and quantitative analysis of multi-components by single-marker method, so as to effectively distinguish Hysspous cuspidatus Boriss. and its adulterants. Methods: High performance liquid chromatography (HPLC) was performed on an Agilent XDB C18 (250 mm×4.6 mm, 5 μm) column with acetonitrile (A)-0.1% formic acid solution as the mobile phase with gradient elution at a flow rate of 1.0 mL · min-1, a column temperature of 30 ℃, a detection wavelength of 280 nm, and an injection volume of 20 μL. The chemical constituents were analysed, and the fingerprinting method was established, combined with similarity evaluation, CA, PCA, OPLS-DA to screen and confirm the differentiation markers. TOPSIS was used to evaluate the different batches of Hysspous cuspidatus Boriss and its adulterants from different places, and to screen the best place of origin. Rosemarinic acid was used to evaluate different bathes of Hysspous cuspidatus Boriss and its adulterants. Using rosemarinic acid as the internal reference, the QAMS method of nine differential quality markers was established. Results: The fingerprints of 22 batches of herbs were established, and a total of 51 common peaks were identified. Fifteen peaks were recognized by comparison with the reference standards, which were neochlorogenic acid, chlorogenic acid, caffeic acid, ferulic acid, quercetin-3-O-β-D-glucopyranosiduronic acid, rosemarinic acid, salvianolic acid B, lignocerotoxin, methyl rosemarinic acid, ethyl rosemarinic acid, β-sitosterol, coriandrin, kaempferol, trichothecenes, and hawthorn acid. The similarities between the samples and reference fingerprint ranged from 0.509 to 0.996. CA classified 22 batches samples into 3 major categories. PCA showed some differences in the chemical compositions of Hysspous cuspidatus Boriss and its adulterants with main differential components of 5. OPLS-DA screened out 12 possible differential quality markers. QAMS was established for nine differential markers. The linearity of the nine differential markers was good, and the RSDs of the precision, stability, repeatability and durability test determinations were all<3.0%, and the average spiked recoveries ranged from 97.5% to 103.1%, with the RSDs all<2.0%. There was no significant difference between the results of QAMS and ESM using rosemarinic acid as the internal reference material. Conclusion: The fingerprint and QAMS method for simultaneously measuring the content of nine components are accurate and reliable to evaluate the quality of Hysspous cuspidatus Boriss.
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