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<p>Lycopodii Herba (LH), the dried whole plant of <i>Lycopodium japonicum</i> Thunb., is frequently confused with other plants within Lycopodiaceae family or other morphologically similar herbs, such as <i>Palhinhaea cernuum</i> L. or <i>Selaginella tamariscina</i>. In this study, the traditional microscopic identification and two high-performance thin-layer chromatography (HPTLC) methods were used to identify the authentic species <i>L. japonicum</i> against two adulterants. By using HPTLC coupled with electrospray ionization‒mass spectrometry (HPTLC‒ESI‒MS) technique, four characteristic zones found in the two HPTLC chromatograms were rapidly identified as <i>N</i>-demethyl-α-obscurine, α-obscurine, β-obscurine, and 9-octadecenoic acid. Among them, <i>N</i>-demethyl-α-obscurine was isolated by separation guided with TLC and structurally elucidated by nuclear magnetic resonace (NMR) data, α-obscurine was verified by comparison with the reference substance. In the microscopic image of LH, the moniliform thickened anticlinal walls of the epidermal cells were found as a typical characteristic differing from the other two species. The research results demonstrated that the traditional microscopic identification method and HPTLC‒MS are both effective and reliable for identification of the authenticity of LH.</p>

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Authenticity of Lycopodii Herba from its adulterant species by high-performance thin-layer chromatography and microscopic identification

  • 期刊论文

  • 发表日期：

    2025年7月31日

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  • Anlu Ma,
  • Chengcheng Ma,
  • Lihua Gu,
  • Lihong Wu,
  • Li Yang,
  • Zhengtao Wang

摘要

Lycopodii Herba (LH), the dried whole plant of Lycopodium japonicum Thunb., is frequently confused with other plants within Lycopodiaceae family or other morphologically similar herbs, such as Palhinhaea cernuum L. or Selaginella tamariscina. In this study, the traditional microscopic identification and two high-performance thin-layer chromatography (HPTLC) methods were used to identify the authentic species L. japonicum against two adulterants. By using HPTLC coupled with electrospray ionization‒mass spectrometry (HPTLC‒ESI‒MS) technique, four characteristic zones found in the two HPTLC chromatograms were rapidly identified as N-demethyl-α-obscurine, α-obscurine, β-obscurine, and 9-octadecenoic acid. Among them, N-demethyl-α-obscurine was isolated by separation guided with TLC and structurally elucidated by nuclear magnetic resonace (NMR) data, α-obscurine was verified by comparison with the reference substance. In the microscopic image of LH, the moniliform thickened anticlinal walls of the epidermal cells were found as a typical characteristic differing from the other two species. The research results demonstrated that the traditional microscopic identification method and HPTLC‒MS are both effective and reliable for identification of the authenticity of LH.