Simultaneous Determination of Rutin, Isoquercetin and Quercetin Flavonoids in Nelumbo nucifera by High-performance Liquid Chromatography Method

  • P Naveen Analytical Research and Development, Vidya Herbs Pvt. Ltd, Bengaluru, Karnataka, India
  • HB Lingaraju Phytochemistry Lab, Vidya Herbs Pvt., Ltd., Bengaluru, Karnataka, India
  • Anitha A Analytical Research and Development, Vidya Herbs Pvt. Ltd, Bengaluru, Karnataka, India
  • K Shyam Prasad Phytochemistry Lab, Vidya Herbs Pvt., Ltd., Bengaluru, Karnataka, India
Keywords: International Conference on Harmonisation, Linearity, Precision, Recovery, Limit of detection, Limit of quantification

Abstract

 Objective: The present study was investigated to provide a documentary evidence for the determination of rutin, isoquercetin and quercetin flavonoids from the flora of Nelumbo nucifera by reversed-phase high-performance liquid chromatography (RP-HPLC). Materials and Methods: RP-HPLC analysis was performed by gradient elution with a low-pressure gradient using 0.5% acetic acid: acetonitrile as a mobile phase with a flow rate of 1.0 ml/min. The separation was done at 26°C using a Kinetex XB-C18 column as stationary phase and the detection wavelength at 356 nm. The proposed method was validated as per International Conference on Harmonisation guidelines with respect to specificity, linearity, precision, accuracy, limit of detection (LOD) and limit of quantification (LOQ). Results: The validated results were within the acceptable limits. In specificity, the retention time of rutin, isoquercetin and quercetin peak in the sample was matched with the reference standard peak and showed good resolution. An excellent linearity was obtained with correlation coefficient (r) higher than 0.999. In precision, the repeatability and intermediate showed <1.0% of % relative standard deviation of peak area percentage indicating high precision and accurate. The recovery rate for rutin, isoquercetin and quercetin was between 99.85%–101.37%, 101.90%–103.24%,and 101.74%–106.73%, respectively. The lower LOD and LOQ of rutin, isoquercetin and quercetin enable the detection and quantitation of these flavonoids in N. nucifera at low concentrations. Conclusion: The developed analytical method is convenient for the determination of flavonoids content in herbal drugs.

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High‑performance liquid chromatograms of blank, standard and sample. The retention time of rutin, isoquercetin and quercetin reference standard and sample (LFE) peak were found to be at 3.748, 4.557, 11.465 and 3.756, 4.562, 11.496 respectively
Published
2017-07-24
How to Cite
1.
Naveen P, Lingaraju H, A A, Prasad KS. Simultaneous Determination of Rutin, Isoquercetin and Quercetin Flavonoids in Nelumbo nucifera by High-performance Liquid Chromatography Method. ijpi [Internet]. 24Jul.2017 [cited 27Jan.2023];7(2):94-00. Available from: https://jpionline.org/index.php/ijpi/article/view/188