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Simple, efficient and economical methods for isolation and estimation of novel isoflavone using RP-HPLC.

Afroze Alam1, Kamlesh Kumar Naik2, Navneet Kumar Upadhaya1

  • 1School of Pharmaceutical Sciences, Shoolini University, Bajhol, Solan, 173229, Himachal Pradesh, India.

Methodsx
|March 15, 2017
PubMed
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A simplified method isolates a novel isoflavone from Iris kashmiriana. This commercially viable procedure avoids chromatography, yielding a highly pure compound (98.9%) for potential applications.

Area of Science:

  • Phytochemistry
  • Natural Product Chemistry
  • Pharmacognosy

Background:

  • Iris kashmiriana is a plant species with potential bioactive compounds.
  • Isoflavones are a class of flavonoids with diverse biological activities.
  • Traditional isolation methods often involve complex chromatographic techniques.

Purpose of the Study:

  • To develop a simplified, commercially viable procedure for isolating bioactive isoflavones from Iris kashmiriana.
  • To characterize the isolated isoflavone using spectroscopic methods.
  • To determine the purity of the isolated compound.

Main Methods:

  • Direct isolation using solvent extraction (petroleum ether, methanol) and successive fractionation (toluene, chloroform, ethyl acetate, n-butanol).
  • Acid hydrolysis of the n-butanol fraction to yield the isoflavone.
Keywords:
Iris kashmerianaIsoflavoneQuantitative estimationRP-HPLCSpectroscopic chracterization

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  • Purification and characterization using Thin Layer Chromatography (TLC), melting point, UV spectroscopy, Infrared (IR) spectroscopy, Nuclear Magnetic Resonance (NMR) spectroscopy (¹H and ¹³C), and Mass Spectrometry.
  • Quantitative analysis using Reverse-Phase High-Performance Liquid Chromatography (RP-HPLC).

    • Main Results:

    • A previously undescribed isoflavone was successfully isolated from Iris kashmiriana.
    • The isolation procedure was simplified, avoiding chromatographic techniques.
    • The isolated isoflavone achieved a purity of 98.9% as determined by RP-HPLC.
    • Comprehensive spectroscopic data confirmed the structure of the isoflavone.

    Conclusions:

    • A novel, simplified, and commercially viable method for isolating isoflavones from Iris kashmiriana has been established.
    • The developed method offers an efficient alternative to traditional isolation techniques.
    • The high purity and confirmed structure of the isolated isoflavone suggest potential for further pharmacological investigation.