Quantification of the secondary metabolites by HPTLC, analgesic and antipyretic activity evaluation of Ficus racemosa L. leaves

  • Amit Kumar
  • Amrita Mishra
  • Arun Kumar Mishra
  • Harpreet SinghEmail author
Research Article


There have been no reports of simultaneous quantification of kaempferol and quercetin in Ficus racemosa L. Objective of the present work included to perform extraction using petroleum ether (PEE) and ethanol (EE), to perform preliminary phytochemical test, quantitative estimation of phytochemicals and simultaneous estimation of kaempferol and quercetin in Ficus racemosa L. using high performance thin layer chromatography (HPTLC) method and to assess the analgesic and antipyretic activity of the extracts. PEE and EE obtained from cold maceration technique. Extracts were subjected to qualitative test and estimated quantitatively. Extracts were applied on silica gel G 60 F254 plate; solvent-Toluene: Chloroform: Acetone 4.5:4.5:1 (v/v) and scanned at 254 nm. The extracts were also subjected to analgesic activity (hot plate method and tail immersion method) and antipyretic (Brewer’s yeast induced pyrexia model and Lipopolysaccharide induced pyrexia mode) activity. Findings of preliminary phytochemical test and quantitative estimation of phytochemicals confirms the presence of flavonoids in both the extracts (PEE and EE). The amount of flavonoids in PEE and EE were found to be 24.58 ± 0.60 mg/100 g and 21.01 ± 0.58 mg/100 g respectively. HPTLC system resulted well resolved bands for quercetin (Rf 0.38) and kaempferol (Rf 0.55). The validated HPTLC method was found suitable for kaempferol and quercetin quantification in Ficus racemosa L. Hot plate reaction time response was improved in a dose-dependent manner in a group of rats treated with PEE suggesting its analgesic potential. In view of analgesic potential by tail immersion method, PEE (400 mg/kg b.w.) showed no significant difference in tail flick reaction time as shown by standard treated animal (Aspirin 100 mg/kg b.w.). The findings of antipyretic activity suggested that PEE (400 mg/kg b.w.) have no significant difference in treating pyrexia when compared with Paracetamol (100 mg/kg). PEE (400 mg/kg b.w.) was found to be most potent. It may be due to the presence of comparatively high amount of flavonoids in PEE confirmed by the preliminary phytochemical test, quantitative estimation of phytochemicals and HPTLC quantification the PEE was found to be more effective when compared to EE.


Kaempferol Quercetin Lipopolysaccharide HPTLC quantification 



Authors are thankful to the Administration of IFTM University, Moradabad for the admirable assistance and motivation to complete the lab work.

Compliance with ethical standards

Ethical statement

The experimental protocols were approved (Ref 2016/839/ac/MPH/31) by the Institutional Animal Ethics Committee (IAEC) constituted under Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA-837/ac/2004) guidelines.

Conflict of interest

This manuscript described has not been published before; not under consideration for publication anywhere else; and has been approved by all co-author.


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Copyright information

© Institute of Korean Medicine, Kyung Hee University and Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  • Amit Kumar
    • 1
  • Amrita Mishra
    • 1
  • Arun Kumar Mishra
    • 1
  • Harpreet Singh
    • 1
    Email author
  1. 1.Central Facility of Instrumentation, School of Pharmaceutical SciencesIFTM UniversityMoradabadIndia

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