青海地区藏药甲地然果(椭圆叶花锚)HPLC指纹图谱和5个主成分含量测定研究

A Study on HPLC Fingerprinting and Quantitative Analysis of Five Major Components in Jia Di Ran Guo(Halenia elliptica D.Don) from Qinghai Tibetan Medicine Region

目的:建立椭圆叶花锚HPLC指纹图谱与含量测定方法。方法:采用Inspire C18(4.6 mm×250 mm,5μm)色谱柱进行分离,乙腈-甲醇-水(含0.03%冰乙酸)系统梯度洗脱,检测波长254 nm,流速1.0 mL·min^(-1),进样量10μL,柱温35℃。采用以上色谱条件对22批椭圆叶花锚样品进行指纹图谱分析,标定了25个共有峰,经对照标品比对,指认出其中5个主要成分的色谱峰,并对这5个成分进行了含量测定。结果:5种主要成分依次为去甲氧基花锚苷、花锚苷、獐牙菜苦苷、异荭草苷、1-羟基-3,4,5-三甲氧基口山酮,含量分别为11.2692~20.4006 mg·g^(-1)、5.0018~11.7832 mg·g^(-1)、3.2782~15.3578 mg·g^(-1)、1.1517~4.5183 mg·g^(-1)、1.4075~6.9511 mg·g^(-1)。22批椭圆叶花锚样品指纹图谱和对照指纹图谱相似度在0.908~0.991之间,各批次药材成分种类基本一致。采用SPSS 25.0与SIMCA 14.1软件对椭圆叶花锚样品5种成分含量进行聚类分析、PCA分析和OPLS-DA分析,可将样品分为3类,椭圆叶花锚差异的标志性成分为花锚苷、去甲氧基花锚苷和獐牙菜苦苷。结论:该方法具有简便、专属性强、稳定、可重复的特点,可为椭圆叶花锚药材的鉴别与质量评价提供参考。

Objective To establish HPLC fingerprint and content determination method of Halenia elliptica D.Don.Methods Separation was achieved using an Inspire C18 column(4.6 mm × 250 mm,5 μ m).With a gradient elution system of acetonitrile-methanol-water(containing 0.03% glacial acetic acid).Detection was performed at a wavelength of 254 nm.With a flow rate of 1.0 mL·min^(-1),an injection volume of 10 μL,and a column temperature of 35℃.The established chromatographic conditions were applied to analyze the fingerprint of 22batches of Halenia elliptica D.Don samples.Twenty-five common peaks were identified through reference standard comparison.Among them,five major components were recognized,and their contents were determined.Results The five major components were identified as deoxyloganin,loganin,swertiamarin,isochlorogenic acid,and1-hydroxy-3,4,5-trimethoxyanthraquinone,with contents ranging from 11.2692 mg·g^(-1) to 20.4006 mg·g^(-1),5.0018 mg·g^(-1) to 11.7832 mg·g^(-1),3.2782 mg·g^(-1) to 15.3578 mg·g^(-1),1.1517 mg·g^(-1) to 4.5183 mg·g^(-1),and 1.4075mg·g^(-1) to 6.9511 mg·g^(-1),respectively.The fingerprint similarity between the chromatographic profiles of 22batches of Halenia elliptica D.Don samples and the reference fingerprint ranged from 0.908 to 0.991,indicating a high consistency in the composition of medicinal components across different batches.Cluster analysis,PCA analysis,and OPLS-DA analysis conducted using SPSS 25.0 and SIMCA 14.1 software categorized the samples into three groups.The discriminative components contributing to the differences in Halenia elliptica were identified as loganin,deoxyloganin,and swertiamarin.Conclusion The proposed method,characterized by its simplicity,specificity,stability,and reproducibility,can serve as a valuable reference for the identification and quality assessment of Halenia elliptica D.Don medicinal materials.