Development and Validation of an HPLC/PDA Methodology for Assessing the Quality of Folium Perilla Frutescens under Diverse Soil Conditions and Growth Stages Based on the Bioactive Compound Rosmarinic Acid
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Abstract
Perilla frutescens (L.), renowned for its medicinal properties such as lowering blood uric acid, anti-inflammatory effects, and anti-allergy effects, contains a variety of beneficial compounds including phenolics, flavonoids, and essential oils. Among these, rosmarinic acid stands out for its remarkable ability to reduce blood uric acid levels. The study has introduced an innovative approach to extract and quantify rosmarinic acid from Perilla leaves, employing a straightforward extraction method with quick analysis. The optimized extraction method involves using ultrasound for 10 minutes with 70% ethanol. For chromatographic analysis, a Phenomenex Luna C18 column (250 mm x 4.6 mm, 5 µm) was utilized with a mobile phase composed of acetonitrile and 0.1% formic acid in a ratio of 27:73 (v/v), flowing at 1.2 mL/min for 7 minutes. This methodology underwent thorough validation following the Association Of Official Analytical Collaboration guidelines, demonstrating a limit of quantification of 0.1 µg/mL. Furthermore, it exhibits excellent linearity within the concentration range of 10-100 µg/mL, supported by a high correlation coefficient of 0.9988. The procedure is applied to quantitatively determine the rosmarinic acid contents in Perilla leaves across 17 provinces in Vietnam, revealing Ho Chi Minh City to have the most abundant concentration of this compound. Research on the growth stages of Perilla frutescens has revealed that the flowering stage exhibits the highest levels of rosmarinic acid. These findings are pivotal for evaluating the quality of raw materials and optimizing the use of Perilla frutescens as a medicinal herb.
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