Comparative Analysis of Rutin Content in Some Egyptian Plants: A Validated RP-HPLC-DAD Approach

Abstract

Aims: To quantify the bioflavonoid rutin in three Egyptian plant families extracts in a comparative study and to prove the accuracy, precision, linearity and reproducibility of the used method.    Study Design: Development of RP-HPLC-DAD method. Rutin analysis in 29 plant extracts. Method validation using different parameters. Place and Duration of Study: Department of Pharmacognosy, Faculty of Pharmacy, Ain Shams University, Cairo, Egypt. The study was performed in 11 months. Methodology: A method was developed using reversed phased-high performance liquid chromatography coupled with diode array detector (RP-HPLC-DAD) and rutin as an analytical marker. Twenty-nine extracts from different Egyptian plants belonging to three families (Lythraceae, Lamiaceae and Asteraceae) were investigated for their rutin content. The method was then validated for accuracy, repeatability, precision, detection, quantification limits, linearity, and range parameters. Results: The content of rutin (mg g-1 extract) was highest in Punica granatum bark (158.29), followed by Melissa officinalis leaves (133.09), Lagerstroemia tomentosa flowers (120.16) and leaves (118.17). The lowest rutin content (4.4 mg g-1 extract) was found in Lagerstroemia speciosa bark. The calibration regression equation was y = 7390x + 15.81 showing a correlation coefficient (r) of 0.9995, with best linearity in the range of 0.01-0.1 mg mL-1. The detection and quantification limit values were 0.0031 and 0.0093 mg mL-1, respectively, confirming the quantification method sensitivity. A recovery value of 100.17% indicates the best method accuracy. Conclusion: The applied method was simple, precise, accurate, and proved successful for rutin determinations in different extracts for the first time in the selected plants, declaring regional variation in the phytoconstituents content of the Egyptian chemotypes and proved that Lythraceae plants were the richest in rutin, with Punica granatum bark extract showing the highest values. The method can be applied for the plants routine quality control analyses and the traces analysis of rutin in complex samples.