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<p><i>Nigella sativa</i> L., is valued for its pharmacological properties, largely attributed to its seeds and their major bioactive compound, thymoquinone. However, limited standardized formulations exist, likely due to challenges in efficient extraction and metabolite stability. In this study, thymoquinone was identified and isolated using preparative column chromatography and confirmed through HPLC analysis. The eluted fractions were allowed to evaporate at ambient temperature, yielding a concentrated yellowish thymoquinone extract. Fractions collected in test tubes 21–35 contained the highest amount of thymoquinone (0.71 ± 0.017&#xa0;mg), followed by fractions 36–50 (0.019 ± 0.001&#xa0;mg). HPLC results confirmed a purity of 98%. The use of preparative column chromatography with optimized solvent combinations, supported by TLC verification, proved to be an effective method for large-scale isolation of thymoquinone.</p>

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Preparative Column Chromatography for the Isolation and Identification of Thymoquinone from Nigella (Nigella sativa L.) Seeds

  • 期刊论文

  • 发表日期：

    2026年1月26日

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  • Y. Ravi,
  • P. I. Vethamoni,
  • S. N. Saxena,
  • S. S. Meena,
  • N. Ashoka,
  • Sharda Choudhary,
  • Ravindra Singh,
  • Arvind Kumar Verma,
  • P. Dhamotharan,
  • Vinay Bhardwaj

摘要

Nigella sativa L., is valued for its pharmacological properties, largely attributed to its seeds and their major bioactive compound, thymoquinone. However, limited standardized formulations exist, likely due to challenges in efficient extraction and metabolite stability. In this study, thymoquinone was identified and isolated using preparative column chromatography and confirmed through HPLC analysis. The eluted fractions were allowed to evaporate at ambient temperature, yielding a concentrated yellowish thymoquinone extract. Fractions collected in test tubes 21–35 contained the highest amount of thymoquinone (0.71 ± 0.017 mg), followed by fractions 36–50 (0.019 ± 0.001 mg). HPLC results confirmed a purity of 98%. The use of preparative column chromatography with optimized solvent combinations, supported by TLC verification, proved to be an effective method for large-scale isolation of thymoquinone.