School of Basic and Applied Sciences, Nirwan University Jaipur, Rajasthan 303305, India; Innovation Centre, Mane Kancor Ingredients Private Limited, Kochi, Kerala 683573, India.
School of Basic and Applied Sciences, Nirwan University Jaipur, Rajasthan 303305, India.
J Chromatogr B Analyt Technol Biomed Life Sci. 2024 Oct 1;1246:124277. doi: 10.1016/j.jchromb.2024.124277. Epub 2024 Aug 21.
The aim of this study was to develop and validate a densitometric High-Performance Thin-Layer Chromatography (HPTLC) method for the simultaneous quantification of quercetin (Q) and kaempferol (K) in Hibiscus mutabilis leaf extracts. The analyses were performed on silica gel 60 F254 plates using a mobile phase composed of toluene, formic acid, and ethyl acetate (6:0.4:4, v/v/v). Detection was carried out at a wavelength of 272 nm using a deuterium and tungsten light source. The method exhibited excellent linearity over the concentration range of 100-600 ng/spot for quercetin and 500-3000 ng/spot for kaempferol, with determination coefficients (r) of 0.9989 and 0.9973, respectively. The method showed no interferences from the plant matrix. The relative standard deviation (RSD) values for intra- and inter-day precision were less than 2% for both flavonoids. Recovery rates ranged from 97.69% to 99.20% for quercetin and from 89.91% to 95.87% for kaempferol. The limits of detection (LOD) were 190.23 ng/spot for quercetin and 187.23 ng/spot for kaempferol, while the limits of quantification (LOQ) were 570.10 ng/spot for quercetin and 566.12 ng/spot for kaempferol. This validated HPTLC method is reliable, precise, and accurate, making it suitable for the quality control of Hibiscus mutabilis leaf extracts. The study's findings can be broadly applied to the quality control of herbal products, pharmacological research, and the development of nutraceuticals. The method's ability to provide rapid and accurate quantification makes it an invaluable tool for researchers across various disciplines.
本研究旨在开发和验证一种高效薄层色谱（HPTLC）密度测定法，用于同时定量分析变色芙蓉叶提取物中的槲皮素（Q）和山奈酚（K）。分析在硅胶 60 F254 板上进行，使用甲苯、甲酸和乙酸乙酯（6:0.4:4，v/v/v）组成的流动相。检测波长为 272nm，使用氘钨灯光源。该方法在槲皮素的浓度范围为 100-600ng/点和山奈酚的浓度范围为 500-3000ng/点时表现出优异的线性关系，测定系数（r）分别为 0.9989 和 0.9973。该方法显示出对植物基质无干扰。两种黄酮类化合物的日内和日间精密度的相对标准偏差（RSD）值均小于 2%。槲皮素的回收率范围为 97.69%-99.20%，山奈酚的回收率范围为 89.91%-95.87%。槲皮素的检测限（LOD）为 190.23ng/点，山奈酚的检测限（LOD）为 187.23ng/点；槲皮素的定量限（LOQ）为 570.10ng/点，山奈酚的定量限（LOQ）为 566.12ng/点。该经验证的 HPTLC 方法可靠、精确且准确，适用于变色芙蓉叶提取物的质量控制。该研究结果可广泛应用于草药产品的质量控制、药理学研究和营养保健品的开发。该方法能够快速准确地定量，是各学科研究人员的宝贵工具。
