摘要
An efficient, sensitive, accurate and rapid analytical ultra-fast liquid chromatography （UFLC） method for quality evaluations ofPyrrosia petiolosa （Christ） Ching from 20 regions of China was developed in this study. Ten marker compounds were simultaneously quantified, including 5-caffeoylquinic acid （5-CQA）, 3-caffeoylquinic acid （3-CQA）, 4-caffeoylquinic acid （4-CQA）, 1-caffeoylquinic acid （1-CQA）, 3,5-dicaffeoylquinic acid （3,5-diCQA）, 4,5-dicaffeoylquinic acid （4,5-diCQA）, 3,4-dicaffeoylquinic acid （3,4-diCQA）, astragalin, kaempferol-3,7-di-O-glucoside and （±）eriodictyol-7-O-β-D-glucuronide. Chromatography was performed on a Kromasil 100-2.5C18 （100 mm×2.1 mm, 2.5 μm） C18 column with gradient elution. The mobile phases consisted of 0.1% formic acid/water （A） and 0.1% formic acid/methanol （B）. The detection wavelength was set at 326 nm and the flow rate was 0.4 mL/min. Ten components were separated well with good linearity （r2〉0.9998）, precision, repeatability, stability. The recovery was in the range of 99.08%-102.77%. The results showed that the content determination using RP-UFLC-DAD fingerprint technique provides an efficient, sensitive, accurate and rapid analytical method for quality assessment ofP. petiolosa （Christ） Ching. Cluster analysis and principal components analysis were successfully applied to analyze 20 samples, the results revealed that the method was efficient and authentic to distinguish producing areas and the source of P. petiolosa （Christ） Ching. Keywords： Pyrrosiapetiolosa （Christ） Ching, Caffeoylquinic acids, Flavonoids, Multicomponent determination, UFLC
在本研究中,建立了高效、灵敏、准确的超快速液相色谱法同时快速分析测定有柄石韦中10个指标成分(新绿原酸、绿原酸、隐绿原酸、1-咖啡酰奎宁酸、3,5-二咖啡酰奎宁酸、4,5-二咖啡酰奎宁酸、3,4-二咖啡酰奎宁酸、紫云英苷、圣草酚-7-O-β-D-吡喃葡萄糖醛酸苷、山奈酚-3,7-双-O-葡萄糖苷)的含量。采用了Kromasil 100-2.5C18(100 mm×2.1 mm,2.5μm)C18快速色谱柱,以0.1%甲酸水和0.1%甲酸甲醇溶液为洗脱剂洗脱,检测波长为326 nm,流速为0.4 mL/min。10个成分在此色谱条件下能达到基线分离,标准曲线线性关系良好(r 2>0.9998),日内与日间精密度、重复性、稳定性均符合实验要求,加样回收率范围为99.1%–104.5%。对20批不同来源的有柄石韦样品进行了含量测定。此研究为有柄石韦中多成分含量测定提供了高效、灵敏、准确的快速分析方法。此外,还对20份样品进行了聚类分析和主成份分析,结果显示此方法可以很好地区分20份样品的来源及资源分布情况。
基金
Study of Safety Testing Techniques and Standards on New Traditional Chinese Drug(National Key Science and Technology Special Projects,Grant No.2014ZX09304307-001-001)
