高效薄层色谱法同时测定药用植物中多酚类化合物的方法开发与验证
Development and validation of High-performance Thin-layer Chromatography Method for Simultaneous Determination of Polyphenolic Compounds in Medicinal Plants.
作者信息
Jayachandran Nair C V, Ahamad Sayeed, Khan Washim, Anjum Varisha, Mathur Rajani
机构信息
Department of Pharmacology, Delhi Institute of Pharmaceutical Sciences and Research, New Delhi, India.
Department of Pharmacognosy and Phytochemistry, Bioactive Natural Product Laboratory, Faculty of Pharmacy, Jamia Hamdard, New Delhi, India.
出版信息
Pharmacognosy Res. 2017 Dec;9(Suppl 1):S67-S73. doi: 10.4103/pr.pr_122_16.
CONTEXT
Quantitative standardization of plant-based products is challenging albeit essential to maintain their quality.
AIMS
This study aims to develop and validate high-performance thin-layer chromatography (HPTLC) method for the simultaneous determination of rutin (Ru), quercetin (Qu), and gallic acid (Ga) from Linn. (PG) and (L.) Correa. (AM) and correlate with antioxidant activity.
MATERIALS AND METHODS
The stock solution (1 mg/mL) of standard Ru, Qu, and Ga in methanol: Water (1:1) was serially diluted and spotted (5 μL) on slica gel 60 F thin-layer chromatography plates. Toluene: Ethyl acetate: Formic acid: Methanol (3:4:0.8:0.7, v/v/v) was selected as mobile phase for analysis at 254 nm. Hydroalcoholic (1:1) extracts of leaves of PG and AM were fractionated and similarly analyzed. Antioxidant activity was also determined using 2, 2-diphenyl-1-picrylhydrazyl assay.
RESULTS
The developed method was robust and resolved Ru, Qu, and Ga at R 0.08 ± 0.02, 0.76 ± 0.01, and 0.63 ± 0.02, respectively. The intra-day, interday precision, and interanalyst were <2% relative standard deviation. The limit of detection and limit of quantification for Ru, Qu, and Ga were 4.51, 4.2, 5.27, and 13.67, 12.73, 15.98 ng/spot, respectively. Antioxidant activity (Log 50% inhibition) of PG and AM was 4.947 ± 0.322 and 6.498 ± 0.295, respectively.
CONCLUSION
The developed HPTLC method was rapid, accurate, precise, reproducible, and specific for the simultaneous estimation of Ru, Qu, and Ga.
SUMMARY
HPTLC method for simultaneous determination and quantification of Rutin, Quercetin and Gallic acid, is reported for quality control of herbal drugs. A: Aqueous fraction; AM: L. Correa; B: Butanol fraction; C: Chloroform fraction; EA: Ethyl acetate fraction; Ga: Gallic acid; H: Hexane fraction; HA: Hydroalcoholic extract; HPTLC: High-performance thin-layer chromatography; PG: ; Qu: Quercetin; Ru: Rutin.
背景
植物基产品的定量标准化具有挑战性，尽管对于维持其质量至关重要。
目的
本研究旨在开发并验证一种高效薄层色谱（HPTLC）方法，用于同时测定来自[植物名称1]（PG）和[植物名称2]（L.）Correa.（AM）中的芦丁（Ru）、槲皮素（Qu）和没食子酸（Ga），并将其与抗氧化活性相关联。
材料与方法
将芦丁、槲皮素和没食子酸标准品（1mg/mL）的储备液用甲醇：水（1:1）进行系列稀释，取5μL点样于硅胶60F薄层色谱板上。选择甲苯：乙酸乙酯：甲酸：甲醇（3:4:0.8:0.7，v/v/v）作为流动相，在254nm波长下进行分析。对PG和AM叶片的水醇（1:1）提取物进行分馏并进行类似分析。还使用2,2-二苯基-1-苦基肼法测定抗氧化活性。
结果
所开发的方法稳健，分别在Rf值为0.08±0.02、0.76±0.01和0.63±0.02处分离出芦丁、槲皮素和没食子酸。日内、日间精密度以及不同分析人员之间的相对标准偏差均<2%。芦丁、槲皮素和没食子酸的检测限和定量限分别为4.51、4.2、5.27和13.67、12.73、15.98ng/斑点。PG和AM的抗氧化活性（半数抑制浓度的对数值）分别为4.947±0.322和6.498±0.295。
结论
所开发的HPTLC方法快速、准确、精密、可重现且特异，可同时测定芦丁、槲皮素和没食子酸。
总结
报道了用于同时测定和定量芦丁、槲皮素和没食子酸的HPTLC方法，用于草药的质量控制。A：水相部分；AM：[植物名称2]；B：丁醇部分；C：氯仿部分；EA：乙酸乙酯部分；Ga：没食子酸；H：己烷部分；HA：水醇提取物；HPTLC：高效薄层色谱；PG：[植物名称1]；Qu：槲皮素；Ru：芦丁。
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